BACKGROUND:L-3-n-butylphthalide(L-NBP) can inhibit phosphorylation of tau protein and reduce the neurotoxicity of beta-amyloid peptide 1-42(Aβ_(1-42)). OBJECTIVE:To observe the neuroprotective effects of L-NBP on cas...BACKGROUND:L-3-n-butylphthalide(L-NBP) can inhibit phosphorylation of tau protein and reduce the neurotoxicity of beta-amyloid peptide 1-42(Aβ_(1-42)). OBJECTIVE:To observe the neuroprotective effects of L-NBP on caspase-3 and nuclear factor kappa-B(NF-κB) expression in a rat model of Alzheimer's disease. DESIGN,TIME AND SETTING:A cell experiment was performed at the Central Laboratory of Provincial Hospital affiliated to Shandong University between January 2008 and August 2008. MATERIALS:L-NBP(purity>98%) was provided by Shijiazhuang Pharma Group NBP Pharmaceutical Company Limited.Aβ_(1-42),3-[4,5-dimethylthiazolo-2]-2,5 iphenyltetrazolium bromide(MTT),and rabbit anti-Caspase-3 polyclonal antibody were provided by Cell Signaling, USA;goat anti-choactase and rabbit anti-NF-κB antibodies were provided by Santa Cruz,USA. METHODS:Primary cultures were generated from rat basal forebrain and hippocampal neurons at 17 or 19 days of gestation.The cells were assigned into five groups:the control group,the Aβ_(1-42) group(2μmol/ L),the Aβ_(1-42) + 0.1μmol/L L-NBP group,the Aβ_(1-42)+ 1μmol/L L-NBP group,and the Aβ_(1-42)+ 10μmol/L L-NBP group.The neurons were treated with Aβ_(1-42)(2μmol/L) alone or in combination with L-NBP(0.1,1,10μmol/L) for 48 hours.Cells in the control group were incubated in PBS. MAIN OUTCOME MEASURES:Morphologic changes were evaluated using inverted microscopy, viability using the MTT method,and the changes in caspase-3 and NF-κB expression using Western blot. RESULTS:Induction with Aβ_(1-42) for 48 hours caused cell death and soma atrophy,and increased caspase-3 and NF-κB expression(P<0.05).L-NBP blocked these changes in cell morphology, decreased caspase-3 and NF-κB expression(P<0.05),and improved cell viability,especially at the high dose(P<0.05). CONCLUSION:Aβ_(1-42) is toxic to basal forebrain and hippocampal primary neurons;L-NBP protects against this toxicity and inhibits the induction of caspase-3 and NF-κB expression.展开更多
The relation between the expression and activity of MMP-9 in C-reactive protein (CRP)-induced human THP-1 mononuclear cells and the activation of nuclear factor kappa-B (NF-кB) was studied to investigate the possible...The relation between the expression and activity of MMP-9 in C-reactive protein (CRP)-induced human THP-1 mononuclear cells and the activation of nuclear factor kappa-B (NF-кB) was studied to investigate the possible role of CRP in plaque destabilization. Human THP-1 cells were incubated in the presence of CRP at 0 (control group), 25, 50 and 100 μg/mL (CRP groups) for 24 h. In PDTC (a specific NF-кB inhibitor) group, the cells were pre-treated with PDTC at 10 μmol/L and then with 100 μg/mL CRP. The conditioned media (CM) and human THP-1 cells in different groups were harvested. MMP-9 expression in CM and human THP-1 cells was measured by ELISA and Western blotting. MMP-9 activity was assessed by fluorogenic substrates. The expression of NF-кB inhibitor α (IкB-α) and NF-кB P65 was detected by Western blotting and ELISA respectively. The results showed that CRP increased the expression and activity of MMP-9 in a dose-dependent manner in the human THP-1 cells. Western blotting revealed that IкB-α expression was decreased in the cells with the concentrations of CRP and ELISA demonstrated that NF-кB P65 expression in the CRP-induced cells was increased. After pre-treatment of the cells with PDTC at 10 μmol/L, the decrease in IкB-α expression and the increase in NF-кB P65 expression in the CRP-induced cells were inhibited, and the expression and activity of MMP-9 were lowered too. It is concluded that increased expression and activity of MMP-9 in CRP-induced human THP-1 cells may be associated with acti- vation of NF-кB. Down-regulation of the expression and activity of MMP-9 may be a new treatment alternative for plaque stabilization by inhibiting the NF-кB activation.展开更多
Medium-chain fatty acids and their derivatives are natural ingredients that support immunological functions in animals.The effects of glycerol monolaurate(GML)on intestinal innate immunity and associated molecular mec...Medium-chain fatty acids and their derivatives are natural ingredients that support immunological functions in animals.The effects of glycerol monolaurate(GML)on intestinal innate immunity and associated molecular mechanisms were investigated using a chicken embryo model.Sixty-four Arbor Acres broiler embryos were randomly allocated into four groups.On embryonic day 17.5,the broiler embryos were administered with 9 mg of GML,which was followed by a 12-h incubation period and a12-h challenge with 32μg of lipopolysaccharide(LPS).On embryonic day 18.5,the jejunum and ileum were harvested.Results indicated that GML reversed the LPS-induced decline in villus height and upregulated the expression of mucin 2(P<0.05).GML decreased LPS-induced malondialdehyde production and boosted antioxidant enzyme activity(P<0.05).GML alleviated LPS-stimulated intestinal secretion of interleukin(IL)-1β,IL-6,and tumor necrosis factor-a(TNF-a)(P<0.05).GML also normalized LPS-induced changes in the gene expression of Toll-like receptor 4,nuclear factor kappa-B p65(NF-κB p65),cyclooxygenase-2,NOD-like receptor protein 3,IL-18,zonula occludens 1,and occludin(P<0.05).GML enhanced as well the expression of AMP-activated protein kinase a1 and claudin 1(P<0.05).In conclusion,GML improved intestinal morphology and antioxidant status by alleviating inflammatory responses and modulating NF-κB signaling in LPS-challenged broiler embryos.展开更多
Background:Inflammation is an important factor in pathological scarring.The role of neutrophils,one of the most important inflammatory cells,in scar hyperplasia remains unclear.The purpose of this article is to study ...Background:Inflammation is an important factor in pathological scarring.The role of neutrophils,one of the most important inflammatory cells,in scar hyperplasia remains unclear.The purpose of this article is to study the correlation between neutrophil extracellular traps(NETs)and scar hyperplasia and identify a new target for inhibiting scar hyperplasia.Methods:Neutrophils were isolated from human peripheral blood by magnetic-bead sorting.NETs in plasma and scars were detected by enzyme-linked immunosorbent assays(ELISAs),immunofluorescence and flow cytometry.Immunohistochemistry was used to assess neutrophil(CD66B)infiltration in hypertrophic scars.To observe the entry of NETs into fibroblasts we used immunofluorescence and flow cytometry.Results:We found that peripheral blood neutrophils in patients with hypertrophic scars were more likely to form NETs(p<0.05).Hypertrophic scars showed greater infiltration with neutrophils and NETs(p<0.05).NETs activate fibroblasts in vitro to promote their differentiation and migration.Inhibition of NETs with cytochalasin in wounds reduced the hyperplasia of scars in mice.We induced neutrophils to generate NETs with different stimuli in vitro and detected the proteins carried by NETs.We did not find an increase in the expression of common scarring factors[interleukin(IL)-17 and transforming growth factor-β(TGF-β),p>0.05].However,inhibiting the production of NETs or degrading DNA reduced the differentiation of fibroblasts intomyofibroblasts.In vitro,NETs were found to be mediated by Toll-like receptor 9(TLR-9)in fibroblasts and further phosphorylated nuclear factor Kappa-B(NF-κB).We found that IL-6,which is downstream of NF-κB,was increased in fibroblasts.Additionally,IL-6 uses autocrine and paracrine signaling to promote differentiation and secretion.Conclusions:Our experiments found that NETs activate fibroblasts through the TLR-9/NF-κB/IL-6 pathway,thereby providing a new target for regulating hypertrophic scars.展开更多
Heavy infection of the virus leads to overproduction of cytokines. The overproduction of cytokine (cytokines storms) is responsible for the critical cases and deaths of COVID-19. The nuclear factor kappa-B stimulates ...Heavy infection of the virus leads to overproduction of cytokines. The overproduction of cytokine (cytokines storms) is responsible for the critical cases and deaths of COVID-19. The nuclear factor kappa-B stimulates the expression of the genes, which is responsible for cytokines storm and RNA transcription. The COVID-19 virus can be controlled by inhibition of nuclear factor kappa-B. Nuclear factor kappa-B is controlled by inhibition of hydrogen peroxide and inhibitor kappa-B kinase enzyme.展开更多
BACKGROUND:The active form of nuclear factor-kappa B(NF- κB)is involved in the initiation,generation,and development of hepatocellular carcinoma(HCC),and is up-regulated in inflammation-associated malignancies.We inv...BACKGROUND:The active form of nuclear factor-kappa B(NF- κB)is involved in the initiation,generation,and development of hepatocellular carcinoma(HCC),and is up-regulated in inflammation-associated malignancies.We investigated the dynamic expression of NF-κB and its influences on the occurrence of HCC through antiangiogenic(thalidomide) intervention in NF-κB activation. METHODS:Hepatoma models were induced with 2-fluorenyl- acetamide(2-FAA,0.05%)in male Sprague-Dawley rats,and thalidomide(100 mg/kg body weight)was administered intragastrically to intervene in NF-κB activation.The pathological changes in the liver of sacrificed rats were assessed after hematoxylin and eosin staining.NF-κB mRNA was amplified by RT-nested PCR.The alterations of NF-κB and vascular endothelial growth factor(VEGF)expression were analyzed by enzyme-linked immunosorbent assay,immunohistochemistry,and Western blotting. RESULTS:Rat hepatocytes showed denatured,precancerous,and cancerous stages in hepatocarcinogenesis,with an increasing tendency of hepatic NF-κB,NF-κB mRNA,and VEGF expression,and their values in the HCC group were higher than those in controls(P<0.001).In the thalidomide- treated group,the morphologic changes generated only punctiform denaturation and necrosis at the early or middle stages,and nodular hyperplasia or a little atypical hyperplasia at the final stages,with the expression of NF-κB (χ2=9.93,P<0.001)and VEGF(χ2=8.024,P<0.001)lower than that in the 2-FAA group. CONCLUSION:NF-κB is overexpressed in hepatocarcinogenesis and antiangiogenic treatment down-regulates the expression of NF-κB and VEGF,and delays the occurrence of HCC.展开更多
BACKGROUND:Urinary trypsin inhibitor(UTI)inhibits the inflammatory response and protects against ischemia- reperfusion(I/R)injury.The inflammatory response is mediated by nuclear factor-kappa B(NF-κB)and its related ...BACKGROUND:Urinary trypsin inhibitor(UTI)inhibits the inflammatory response and protects against ischemia- reperfusion(I/R)injury.The inflammatory response is mediated by nuclear factor-kappa B(NF-κB)and its related target genes and products such as vascular endothelial cell adhesion molecule and CXC chemokines.We aimed to assess the roles of those mediators in a UTI-treated mouse model of hepatic I/R injury. METHODS:Treatment group 1(UTI given 5 minutes prior to liver ischemia),treatment group 2(UTI given 5 minutes after the anhepatic phase)and a control group were investigated.Blood and liver samples were obtained and compared at 1,3,6 and 24 hours after reperfusion. RESULTS:Attenuation of pathological hepatocellular damage was greater in the treatment groups than in the control group(P<0.05).Compared with the control group, the UTI treatment groups showed significantly lower serum alanine aminotransferase and aspartate aminotransferase levels,decreased myeloperoxidase activity,and reduced NF- κB activation.Also downregulated was the expression of tumor necrosis factor-alpha,cytokine-induced neutrophil chemoattractant,and macrophage inflammatory protein-2 at the mRNA level.P-selectin protein and intercellular adhesion molecule-1 protein expression were also downregulated.In addition,the treatment group 1 showed a better protective effect against I/R injury than the treatment group 2.CONCLUSIONS:UTI reduces NF-κB activation and downregulates the expression of its related mediators, followed by the inhibition of neutrophil aggregation and infiltration in hepatic I/R injury.The protective role of UTI is more effective in prevention than in treatment.展开更多
AIM: To investigate whether NF-κB is activated in human gastric carcinoma tissues and, if so, to study whether there is any correlation between NF-κB activity and heparanase expression in gastric carcinoma.METHODS: ...AIM: To investigate whether NF-κB is activated in human gastric carcinoma tissues and, if so, to study whether there is any correlation between NF-κB activity and heparanase expression in gastric carcinoma.METHODS: NF-κB activation was assayed by immunohistochemical staining in formalin-fixed, paraffin-embedded specimens from 45 gastric carcinoma patients. Electrophoretic mobility shift assay (EMSA) method was used for nuclear protein from these fresh tissue specimens. Heparanase gene expression was quantified using quantitative RT-PCR.RESULTS: The nuclear translocation of RelA (marker of NF-κB activation) was significantly higher in tumor cells compared to adjacent and normal epithelial cells [(41.3±3.52)% vs (0.38±0.22) %, t= 10.993, P= 0.000＜0.05; (41.3±3.52)%vs (0±0.31)%, t = 11.484, P = 0.000＜0.05]. NF-κB activation was correlated with tumor invasion-related clinicopathological features such as lymphatic invasion,pathological stage, and depth of invasion (Z = 2.148,P= 0.032＜0.05; χ2 = 8.758, P= 0.033＜0.05; χ2 = 18.531,P = 0.006＜0.05). NF-κB activation was significantly correlated with expression of heparanase gene (r= 0.194,P= 0.046＜0.05).CONCLUSION: NF-κB RelA (p65) activation was related with increased heparanase gene expression and correlated with poor clinicopathological characteristics in gastric cancers. This suggests NF-κB as a major controller of the metastatic phenotype through its reciprocal regulation of some metastasis-related genes.展开更多
Concomitantly with the increase in the prevalences of overweight/obesity, nonalcoholic fatty liver disease(NAFLD) has worldwide become the main cause of chronic liver disease in both adults and children. Patients with...Concomitantly with the increase in the prevalences of overweight/obesity, nonalcoholic fatty liver disease(NAFLD) has worldwide become the main cause of chronic liver disease in both adults and children. Patients with fatty liver display features of metabolic syndrome(Met S), like insulin resistance(IR), glucose intolerance, hypertension and dyslipidemia. Recently, epidemiological studies have linked obesity, Met S, and NAFLD to decreased bone mineral density and osteoporosis, highlighting an intricate interplay among bone, adipose tissue, and liver. Osteoprotegerin(OPG), an important symbol of the receptor activator of nuclear factor-B ligand/receptor activator of nuclear factor kappa B/OPG system activation, typically considered for its role in bone metabolism, may also play critical roles in the initiation and perpetuation of obesityrelated comorbidities. Clinical data have indicated that OPG concentrations are associated with hypertension, left ventricular hypertrophy, vascular calcification, endothelial dysfunction, and severity of liver damage in chronic hepatitis C. Nonetheless, the relationship between circulating OPG and IR as a key feature of Met S as well as between OPG and NAFLD remains uncertain. Thus, the aims of the present review are to provide the existent knowledge on these associations and to discuss briefly the underlying mechanisms linking OPG and NAFLD.展开更多
AIM: To detect the nuclear factor kappa B (NF-κB) condition in human stage Ⅳ gastric carcinoma patients and to explore the correlation between NF- κB activation and survival of these patients after chemotherapy. ME...AIM: To detect the nuclear factor kappa B (NF-κB) condition in human stage Ⅳ gastric carcinoma patients and to explore the correlation between NF- κB activation and survival of these patients after chemotherapy. METHODS: Expression of NF-κB-p65 was determined by immunohistochemical analysis. Activity of NF- κB DNA-binding in carcinoma tissue was detected by electrophoretic mobility shift assay. Kaplan-Meier survival analysis was performed to show the relation between NF-κB and progression-free survival (PFS) or overall survival (OS) of the patients. RESULTS: The positive expression rate of NF-κB-p65 in 60 gastric cancer tissue samples was 76.7% (46/60). The expression of NF-κB-p65 was reduced in adjacent carcinoma and normal tissue samples. Electrophoretic mobility shift assay (EMSA) analysis showed a strong activation of NF-κB in cancer tissue samples. A survival difference was found in NF- κB-p65 positive and negative patients. NF-κB-p65 expression was negative in cancer tissue samples (n = 14). PFS was 191.40 ± 59.88 d and 152.93 ± 16.99 d, respectively, in patients with positive NF- κB-p65 expression (n = 46) (P = 0.4028). Thesurvival time of patients with negative and positive NF-κB-p65 expression was 425.16 ± 61.61 d and 418.85 ± 42.98 d, respectively (P = 0.7303). Kaplan-Meier analysis showed no significant difference in PFS or OS. The 46 patient tissue which positive NF- κB-p65 expression was found in the tissue samples from the 46 patients whose PFS and OS were 564.89 ± 75.94 d and s 352.37 ± 41.32 d, respectively (P = 0.0165). CONCLUSION: NF-κB is activated in gastric carcinoma tissue, which is related to the OS after chemotherapy.展开更多
AIM: Nuclear factor kappa B (NF-κB) regulates a large number of genes involved in the inflammatory response to critical illnesses, but it is not known if and how NF-κB is activated and intercellular adhesion molecul...AIM: Nuclear factor kappa B (NF-κB) regulates a large number of genes involved in the inflammatory response to critical illnesses, but it is not known if and how NF-κB is activated and intercellular adhesion molecule-1 (ICAM-1)expressed in the gut following traumatic brain injury (TBI).The aim of current study was to investigate the temporal pattern of intestinal NF-κB activation and ICAM-1expression following TBI.METHODS: Male Wistar rats were randomly divided into six groups (6 rats in each group) including controls with sham operation and TBI groups at hours 3, 12, 24, and 72, and on d 7. Parietal brain contusion was adopted using weight-dropping method. All rats were decapitated at corresponding time point and mid-jejunum samples were taken. NF-κB binding activity in jejunal tissue was measured using EMSA. Immunohistochemistry was used for detection of ICAM-1 expression in jejunal samples.RESULTS: There was a very low NF-κB binding activity and little ICAM-1 expression in the gut of control rats after sham surgery. NF-κB binding activity in jejunum significantly increased by 160% at 3 h following TBI (P＜0.05 vs control), peaked at 72 h (500% increase)and remained elevated on d 7 post-injury by 390% increase. Compared to controls, ICAM-1 was significantly up-regulated on the endothelia of microvessels in villous interstitium and lamina propria by 24 h following TBI and maximally expressed at 72 h post-injury (P＜0.001). The endothelial ICAM-1 immunoreactivity in jejunal mucosa still remained strong on d 7 post-injury. The peak of NF-κB activation and endothelial ICAM-1 expression coincided in time with the period during which secondary mucosal injury of the gut was also at their culmination following TBI.CONCLUSION: TBI could induce an immediate and persistent up-regulation of NF-κB activity and subsequent up-regulation of ICAM-1 expression in the intestine.Inflammatory response mediated by increased NF-κB activation and ICAM-1 expression may play an important role in the pathogenesis of acute gut mucosal injury following TBI.展开更多
BACKGROUND:Hepatocellular carcinoma(HCC)is one of the most common malignant tumors.We analyzed the expression of nuclear-transcription factor-kappa B(NF-κB) during hepatocarcinogenesis in order to evaluate its dynami...BACKGROUND:Hepatocellular carcinoma(HCC)is one of the most common malignant tumors.We analyzed the expression of nuclear-transcription factor-kappa B(NF-κB) during hepatocarcinogenesis in order to evaluate its dynamic expression and its clinical value in the development and diagnosis of HCC. METHODS:Hepatoma models were induced by oral administration of 2-acetamidoflurene(2-FAA)to male Sprague-Dawley rats.Morphological changes were observed after hematoxylin and eosin staining.The cellular distribution of NF-κB expression during different stages of cancer development was investigated by immunohistochemistry, and the level of NF-κB expression in liver tissues was quantitatively analyzed by ELISA.The gene fragments of hepatic NF-κB were amplified by nested-polymerase chain reaction assay. RESULTS:Hepatocytes showed vacuole-like degeneration during the early stages,then had a hyperplastic nodal appearance during the middle stages,and finally progressed to tubercles of cancerous nests with high differentiation. The NF-κB-positive material was buff-colored,fine particles localized in the nucleus,and the incidence of NF-κB-positive cells was 81.8%in degeneration,83.3%in precancerous lesions,and 100%in cancerous tissues.All of these values were higher than those in controls(P<0.01). Hepatic NF-κB expression and hepatic NF-κB-mRNA were also higher during the course of HCC development(P<0.01).CONCLUSION:The NF-κB signal transduction pathway is activated during the early stages of HCC development, and its abnormal expression may be associated with the occurrence of HCC.展开更多
Activation of nuclear factor kappa B(NF-κB) is a hallmark of various central nervous system(CNS) pathologies. Neuron-specific inhibition of its transcriptional activator subunit RelA, also referred to as p65, promote...Activation of nuclear factor kappa B(NF-κB) is a hallmark of various central nervous system(CNS) pathologies. Neuron-specific inhibition of its transcriptional activator subunit RelA, also referred to as p65, promotes neuronal survival under a range of conditions, i.e., for ischemic or excitotoxic insults. In macro- and microglial cells, post-lesional activation of NF-κB triggers a growth-permissive program which contributes to neural tissue inflammation, scar formation, and the expression of axonal growth inhibitors. Intriguingly, inhibition of such inducible NF-κB in the neuro-glial compartment, i.e., by genetic ablation of RelA or overexpression of a transdominant negative mutant of its upstream regulator IκBα, significantly enhances functional recovery and promotes axonal regeneration in the mature CNS. By contrast, depletion of the NF-κB subunit p50, which lacks transcriptional activator function and acts as a transcriptional repressor on its own, causes precocious neuronal loss and exacerbates axonal degeneration in the lesioned brain. Collectively, the data imply that NF-κB orchestrates a multicellular program in which κB-dependent gene expression establishes a growth-repulsive terrain within the post-lesioned brain that limits structural regeneration of neuronal circuits. Considering these subunit-specific functions, interference with the NF-κB pathway might hold clinical potentials to improve functional restoration following traumatic CNS injury.展开更多
AIM: To investigate the effect of different dietary fatty acids on hepatic inflammasome activation.METHODS: Wild-type C57BL/6 mice were fed either a high-fat diet or polyunsaturated fatty acid(PUFA)-enriched diet. Pri...AIM: To investigate the effect of different dietary fatty acids on hepatic inflammasome activation.METHODS: Wild-type C57BL/6 mice were fed either a high-fat diet or polyunsaturated fatty acid(PUFA)-enriched diet. Primary hepatocytes were treated with either saturated fatty acids(SFAs) or PUFAs as well as combined with lipopolysaccharide(LPS). The expression of NOD-like receptor protein 3(NLRP3) inflammasome, peroxisome proliferator-activated receptor-γ and nuclear factor-kappa B(NF-κB) was determined by real-time PCR and Western blot. The activity of Caspase-1 and interleukine-1β production were measured.RESULTS: high-fat diet-induced hepatic steatosis was sufficient to induce and activate hepatic NLRP3 inflammasome. SFA palmitic acid(PA) directly activated NLRP3 inflammasome and increased sensitization to LPS-induced inflammasome activation in hepatocytes. In contrast, PUFA docosahexaenoic acid(Dh A) had thepotential to inhibit NLRP3 inflammasome expression in hepatocytes and partly abolished LPS-induced NLRP3 inflammasome activation. Furthermore, a highfat diet increased but PUFA-enriched diet decreased sensitization to LPS-induced hepatic NLRP3 inflammasome activation in vivo. Moreover, PA increased but Dh A decreased phosphorylated NF-κB p65 protein expression in hepatocytes.CONCLUSION:Hepatic NLRP 3 inflammasome activation played an important role in the development of non-alcoholic fatty liver disease. Dietary SFAs and PUFAs oppositely regulated the activity of NLRP3 inflammasome through direct activation or inhibition of NF-κB.展开更多
AIM: To investigate the role of p38 mitogen-activated protein kinase (p38MAPK) in murine experimental autoimmune hepatitis (EAH). METHODS: To induce EAH, the syngeneic S-100 antigen emulsified in complete Freud's ...AIM: To investigate the role of p38 mitogen-activated protein kinase (p38MAPK) in murine experimental autoimmune hepatitis (EAH). METHODS: To induce EAH, the syngeneic S-100 antigen emulsified in complete Freud's adjuvant was injected intraperitoneally into adult male C57Bl/6 mice. Liver injury was assessed by serum ALT and liver histology. The expression and activity of p38 MAPK were measured by Western blot and kinase activity assays. In addition, DNA binding activities of nuclear factor kappa B (NF-κB) were analyzed by electrophoretic mobility shift assay. The effects of SB203580, a specifi c p38 MAPK inhibitor, on liver injuries and expression of proin? ammatory cytokines (interferon-γ, IL-12, IL-1β and TNF-α) were observed. RESULTS: The activity of p38 MAPK and NF-κB was increased and reached its peak 14 or 21 d after the fi rst syngeneic S-100 administration. Inhibition of p38 MAPK activation by SB203580 decreased the activation of NF-κB and the expression of proin? ammatory cytokines. Moreover, hepatic injuries were improved significantly after SB203580 administration. CONCLUSION: p38 MAPK and NF-κB play an important role in an animal model of autoimmune hepatitis (AIH) induced by autoantigens.展开更多
A mouse model of viral encephalitis was induced by intracranial injection of a Coxsackie virus B3 suspension. Quantitative real-time reverse transcription-PCR and western blot assay were applied to detect mRNA and pro...A mouse model of viral encephalitis was induced by intracranial injection of a Coxsackie virus B3 suspension. Quantitative real-time reverse transcription-PCR and western blot assay were applied to detect mRNA and protein expression of intelectin-2 and nuclear factor-kappa B in the viral encephalitis and control groups. Nuclear factor-kappa B and intelectin-2 mRNA and protein expression were significantly increased in mice with viral encephalitis. After intraperitoneal injection of Shuanghuanglian at a dose of 1.5 mg/kg for 5 successive days, intelectin-2 and nuclear factor-kappa B protein and mRNA expression were significantly decreased. To elucidate the relationship between intelectin-2 and nuclear factor-kappa B, mice with viral encephalitis were administered an intracerebral injection of 107 pfu recombinant lentivirus expressing intelectin shRNA. Both protein and mRNA levels of intelectin and nuclear factor-kappa B in brain tissue of mice were significantly decreased. Experimental findings suggest that Shuanghuanglian injection may downregulate nuclear factor-kappa B production via suppression of intelectin production, thus inhibiting inflammation associated with viral encephalitis.展开更多
AIM: To investigate the effect of high homocysteine(Hcy) levels on apolipoprotein E(apoE) expression and the signaling pathways involved in this gene regulation.METHODS: Reverse transcriptase polymerase chain reaction...AIM: To investigate the effect of high homocysteine(Hcy) levels on apolipoprotein E(apoE) expression and the signaling pathways involved in this gene regulation.METHODS: Reverse transcriptase polymerase chain reaction(RT-PCR) and Western blot were used to assess apo E expression in cells treated with various concentrations(50-500 μmol/L) of Hcy. Calcium phosphatetransient transfections were performed in HEK-293 and RAW 264.7 cells to evaluate the effect of Hcy on apoE regulatory elements [promoter and distal multienhancer 2(ME2)]. To this aim, plasmids containing the proximal apoE promoter [(-500/+73)apoE construct] alone or in the presence of ME2 [ME2/(-500/+73)apoE construct] to drive the expression of the reporter luciferase gene were used. Co-transfection experiments were carried out to investigate the downstream effectors of Hcymediated regulation of apoE promoter by using specific inhibitors or a dominant negative form of IKβ. In other co-transfections, the luciferase reporter was under the control of synthetic promoters containing multiple specific binding sites for nuclear factor kappa B(NF-κB), activator protein-1(AP-1) or nuclear factor of activated T cells(NFAT). Chromatin immunoprecipitation(ChI P)assay was accomplished to detect the binding of NF-κB p65 subunit to the apoE promoter in HEK-293 treated with 500 μmol/L Hcy. As control, cells were incubated with similar concentration of cysteine. NF-κB p65 proteins bound to DNA were immunoprecipitated with anti-p65 antibodies and DNA was identified by PCR using primers amplifying the region-100/+4 of the apoE gene. RESULTS: RT-PCR revealed that high levels of Hcy(250-750 μmol/L) induced a 2-3 fold decrease in apoE m RNA levels in HEK-293 cells, while apo E gene expression was not significantly affected by treatment with lower concentrations of Hcy(100 μmol/L). Immunoblotting data provided additional evidence for the negative role of Hcy in apoE expression. Hcy decreased apoE promoter activity, in the presence or absence of ME2, in a dose dependent manner, in both RAW 264.7 and HEK-293 cells, as revealed by transient transfection experiments. The downstream effectors of the signaling pathways of Hcy were also investigated. The inhibitory effect of Hcy on the apo E promoter activity was counteracted by MAPK/ERK kinase 1/2(MEK1/2) inhibitor U0126, suggesting that MEK1/2 is involved in the downregulation of apoE promoter activity by Hcy. Our data demonstrated that Hcy-induced inhibition of apoE took place through activation of NF-κB. Moreover, we demonstrated that Hcy activated a synthetic promoter containing three NF-κB binding sites, but did not affect promoters containing AP-1 or NFAT binding sites. ChI P experiments revealed that NF-κB p65 subunit is recruited to the apoE promoter following Hcy treatment of cells.CONCLUSION: Hcy-induced stress negatively modulates apoE expression via MEK1/2 and NF-κB activation. The decreased apo E expression in peripheral tissues may aggravate atherosclerosis, neurodegenerative diseases and renal dysfunctions.展开更多
AIM: To explore the effects of ketamine on hemo- dynamics, plasma proinflammatory cytokine (TNF-α and IL-6) levels and nuclear factor kappa B (NF-κB) activation during polymicrobial sepsis. METHODS: Male Sprague-Daw...AIM: To explore the effects of ketamine on hemo- dynamics, plasma proinflammatory cytokine (TNF-α and IL-6) levels and nuclear factor kappa B (NF-κB) activation during polymicrobial sepsis. METHODS: Male Sprague-Dawlay rats were subjected to cecal ligation and puncture (CLP) or sham operation. The rats were randomly assigned into four equal groups: sham CLP group, CLP group, ketamine (KT)Ⅰgroup and KTⅡgroup. Thirty minutes before CLP, ketamine (5 mg/kg per hour and 10 mg/kg per hour, respectively) was infused continuously through the left femoral vein cannula in KTⅠgroup or KTⅡgroup. Sham CLP group and CLP group received 0.9% saline only (5 mL/kg per hour). The right femoral artery was cannulated to monitor mean arterial pressure (MAP) and heart rates (HR),and draw blood samples. The proinflammatory cytokine (TNF-α and IL-6) levels of plasma were measured using enzyme-linked immunosorbent assays (ELISA). The hepatic NF-κB activation was determined by Western blot and HPIAS 2000 image analysis system. Twenty hours after CLP, the rats were killed by right femoral artery phlebotomization. RESULTS: CLP produced progressive hypotension, and a first increase followed by a decrease in HR. The hypotension was prevented, and the HR was slightly steady in ketamine treated rats. TNF-α levels of plasma reached a peak value at 2 h after CLP. Ketamine (KT Ⅰgroup or KTⅡgroup) caused a significant decrease compared with CLP group at 2, 5 and 9 h time points after CLP (14.3 ± 1.9 vs 4.3 ± 0.9, 9.7 ± 1.4 vs 4.3 ± 0.9; 9.3 ± 1.5 vs 4.3 ± 0.9, 8.7 ± 1.4 vs 4.3 ± 0.9; 6.0 ± 1.5 vs 5.0 ± 1.7, 5.3 ± 0.8 vs 5.0 ± 1.7; P < 0.01, respectively). The IL-6 levels of plasma firstly ascendedand then descended in CLP group, and reached a peak value at 9 h after CLP. Ketamine (KTⅠgroup or KTⅡ group) caused a significant decrease compared with CLP group at 5, 9 or 20 h after CLP (135.0 ± 52.6 vs 60.0 ± 16.3, 112.5 ± 52.6 vs 60.0 ± 16.3; 410.0 ± 68.7 vs 62.5 ± 12.5, 250.0 ± 28.0 vs 62.5 ± 12.5; 320.0 ± 25.9 vs 52.5 ± 10.1, 215.0 ± 44.6 vs 52.5 ± 10.1; P < 0.05, respectively). The IL-6 levels of plasma in KTⅡgroup were lower than those of KTⅠgroup at 9 h after CLP (250.0 ± 28.0 vs 410.0 ± 68.7; P < 0.05). In addition, CLP increased hepatic NF-κB expression compared with sham CLP. Ketamine suppressed NF-κB activation in a dose-dependent manner at 4 h after CLP (237.7 ± 3.5 vs 246.9 ± 3.1; P < 0.05). CONCLUSION: Ketamine stabilizes the hemodynamics, attenuates the proinflammatory cytokine responses, and inhibits hepatic NF-κB activation. These findings suggest that ketamine has protective effects against polymicrobial sepsis in rats.展开更多
AIM:To investigate genetic diversity of Helicobacter pylori (H.pylori) cell division-related gene A (cdrA) and its effect on the host response.METHODS:Inactivation of H.pylori cdrA,which is involved in cell division a...AIM:To investigate genetic diversity of Helicobacter pylori (H.pylori) cell division-related gene A (cdrA) and its effect on the host response.METHODS:Inactivation of H.pylori cdrA,which is involved in cell division and morphological elongation,has a role in chronic persistent infections.Genetic property of H.pylori cdrA was evaluated using polymerase chain reaction and sequencing in 128 (77 American and 51 Japanese) clinical isolates obtained from 48 and 51 patients,respectively.Enzyme-linked immunosorbent assay was performed to measure interleukin-8 (IL-8) secretion with gastric biopsy specimens obtained from American patients colonized with cdrA-positive or-negative strains and AGS cells cocultured with wild-type HPK5 (cdrA-positive) or its derivative HPKT510 (cdrA-disruptant).Furthermore,the cytotoxin-associated gene A (cagA) status (translocation and phosphorylation) and kinetics of transcription factors [nuclear factor-kappa B (NF-κB) and inhibition kappa B] were investigated in AGS cells co-cultured with HPK5,HPKT510 and its derivative HPK5CA (cagA disruptant) by western blotting analysis with immunoprecipitation.RESULTS:Genetic diversity of the H.pylori cdrA gene demonstrated that the cdrA status segregated into two categories including four allele types,cdrA-positive (allele types;Ⅰand Ⅱ) and cdrA-negative (allele types;Ⅲ and Ⅳ) categories,respectively.Almost all Japanese isolates were cdrA-positive (Ⅰ:7.8% and Ⅱ:90.2%),whereas 16.9% of American isolates were cdrA-positive (Ⅱ) and 83.1% were cdrA-negative (Ⅲ:37.7% and Ⅳ:45.5%),indicating extended diversity of cdrA in individual American isolates.Comparison of each isolate from different regions (antrum and corpus) in the stomach of 29 Americans revealed that cdrA status was identical in both isolates from different regions in 17 cases.However,12 cases had a different cdrA allele and 6 of them exhibited a different cdrA category between two regions in the stomach.Furthermore,in 5 of the 6 cases possessing a different cdrA category,cdrA-negative isolate existed in the corpus,suggesting that cdrA-negative strain is more adaptable to colonization in the corpus.IL-8 secretions from AGS revealed that IL-8 levels induced by a cdrA-disrupted HPKT510 was significantly lower (P < 0.01) compared to wildtype HPK5:corresponding to 50%-60% of those of wild-type HPK5.These data coincided with in vivo data that an average value of IL-8 in biopsy specimens from cdrA-positive and cdrA-negative groups was 215.6 and 135.9 pg/mL,respectively.Western blotting analysis documented that HPKT510 had no effect on CagA translocation and phosphorylation,however,nuclear accumulation of NF-κB was lower by HPKT510 compared to HPK5.CONCLUSION:Colonization by a cdrA-negative or cdrA-dysfunctional strain resulted in decreased IL-8 production and repression of NF-κB,and hence,attenuate the host immunity leading to persistent infection.展开更多
Progranulin is closely related to neuronal survival in a neuroinflammatory mouse model and attenuates inflammatory reactions. Atsttrin is an engineered protein composed of three progranulin fragments and has been show...Progranulin is closely related to neuronal survival in a neuroinflammatory mouse model and attenuates inflammatory reactions. Atsttrin is an engineered protein composed of three progranulin fragments and has been shown to have an effect similar to that of progranulin. Atsttrin has anti-inflammatory actions in multiple arthritis mouse models, and it protects against further arthritis development. However, whether Atsttrin has a role in neuroinflammation remains to be elucidated. In this study, we produced a neuroinflammatory mouse model by intracerebroventricular injection of 1 μL lipopolysaccharide(10 μg/μL). Atsttrin(2.5 mg/kg) was administered via intraperitoneal injection every 3 days over a period of 7 days before intracerebroventricular injection of 1 μL lipopolysaccharide(10 μg/μL). In addition, astrocyte cultures were treated with 0, 100 or 300 ng/mL lipopolysaccharide, with 200 ng/mL Atsttrin simultaneously. Immunohistochemistry, enzyme-linked immunosorbent assay and real-time reverse transcription-polymerase chain reaction were performed to examine the protein and mRNA levels of inflammatory mediators and to assess activation of the nuclear factor kappa B signaling pathway. Progranulin expression in the brain of wild-type mice and in astrocyte cultures was increased after lipopolysaccharide administration. The protein and mRNA expression levels of tumor necrosis factor-α, interleukin-1β and inducible nitric oxide synthase were increased in the brain of progranulin knockout mice after lipopolysaccharide administration. Atsttrin treatment reduced the lipopolysaccharide-induced increase in the protein and mRNA levels of tumor necrosis factor-α, interleukin-1β, matrix metalloproteinase-3 and inducible nitric oxide synthase in the brain of progranulin knockout mice. Atsttrin also reduced the expression of cyclooxygenase-2, inducible nitric oxide synthase and matrix metalloproteinase 3 mRNA in lipopolysaccharide-treated astrocytes in vitro, and decreased the concentration of tumor necrosis factor α and interleukin-1β in the supernatant. Furthermore, Atsttrin significantly reduced the levels of phospho-nuclear factor kappa B inhibitor α in the brain of lipopolysaccharide-treated progranulin knockout mice and astrocytes, and it decreased the expression of nuclear factor kappa B2 in astrocytes. Collectively, our findings show that the anti-neuroinflammatory effect of Atsttrin involves inhibiton of the nuclear factor kappa B signaling pathway, and they suggest that Atsttrin may have clinical potential in neuroinflammatory therapy.展开更多
基金Supported by:the Medicine and Health Scientific Research Projects of Shandong Province,No. 2007HZ065
文摘BACKGROUND:L-3-n-butylphthalide(L-NBP) can inhibit phosphorylation of tau protein and reduce the neurotoxicity of beta-amyloid peptide 1-42(Aβ_(1-42)). OBJECTIVE:To observe the neuroprotective effects of L-NBP on caspase-3 and nuclear factor kappa-B(NF-κB) expression in a rat model of Alzheimer's disease. DESIGN,TIME AND SETTING:A cell experiment was performed at the Central Laboratory of Provincial Hospital affiliated to Shandong University between January 2008 and August 2008. MATERIALS:L-NBP(purity>98%) was provided by Shijiazhuang Pharma Group NBP Pharmaceutical Company Limited.Aβ_(1-42),3-[4,5-dimethylthiazolo-2]-2,5 iphenyltetrazolium bromide(MTT),and rabbit anti-Caspase-3 polyclonal antibody were provided by Cell Signaling, USA;goat anti-choactase and rabbit anti-NF-κB antibodies were provided by Santa Cruz,USA. METHODS:Primary cultures were generated from rat basal forebrain and hippocampal neurons at 17 or 19 days of gestation.The cells were assigned into five groups:the control group,the Aβ_(1-42) group(2μmol/ L),the Aβ_(1-42) + 0.1μmol/L L-NBP group,the Aβ_(1-42)+ 1μmol/L L-NBP group,and the Aβ_(1-42)+ 10μmol/L L-NBP group.The neurons were treated with Aβ_(1-42)(2μmol/L) alone or in combination with L-NBP(0.1,1,10μmol/L) for 48 hours.Cells in the control group were incubated in PBS. MAIN OUTCOME MEASURES:Morphologic changes were evaluated using inverted microscopy, viability using the MTT method,and the changes in caspase-3 and NF-κB expression using Western blot. RESULTS:Induction with Aβ_(1-42) for 48 hours caused cell death and soma atrophy,and increased caspase-3 and NF-κB expression(P<0.05).L-NBP blocked these changes in cell morphology, decreased caspase-3 and NF-κB expression(P<0.05),and improved cell viability,especially at the high dose(P<0.05). CONCLUSION:Aβ_(1-42) is toxic to basal forebrain and hippocampal primary neurons;L-NBP protects against this toxicity and inhibits the induction of caspase-3 and NF-κB expression.
文摘The relation between the expression and activity of MMP-9 in C-reactive protein (CRP)-induced human THP-1 mononuclear cells and the activation of nuclear factor kappa-B (NF-кB) was studied to investigate the possible role of CRP in plaque destabilization. Human THP-1 cells were incubated in the presence of CRP at 0 (control group), 25, 50 and 100 μg/mL (CRP groups) for 24 h. In PDTC (a specific NF-кB inhibitor) group, the cells were pre-treated with PDTC at 10 μmol/L and then with 100 μg/mL CRP. The conditioned media (CM) and human THP-1 cells in different groups were harvested. MMP-9 expression in CM and human THP-1 cells was measured by ELISA and Western blotting. MMP-9 activity was assessed by fluorogenic substrates. The expression of NF-кB inhibitor α (IкB-α) and NF-кB P65 was detected by Western blotting and ELISA respectively. The results showed that CRP increased the expression and activity of MMP-9 in a dose-dependent manner in the human THP-1 cells. Western blotting revealed that IкB-α expression was decreased in the cells with the concentrations of CRP and ELISA demonstrated that NF-кB P65 expression in the CRP-induced cells was increased. After pre-treatment of the cells with PDTC at 10 μmol/L, the decrease in IкB-α expression and the increase in NF-кB P65 expression in the CRP-induced cells were inhibited, and the expression and activity of MMP-9 were lowered too. It is concluded that increased expression and activity of MMP-9 in CRP-induced human THP-1 cells may be associated with acti- vation of NF-кB. Down-regulation of the expression and activity of MMP-9 may be a new treatment alternative for plaque stabilization by inhibiting the NF-кB activation.
基金supported by the National Natural Science Foundation of China(32272910)the Natural Science Foundation of Shandong Province(ZR2020MC170).
文摘Medium-chain fatty acids and their derivatives are natural ingredients that support immunological functions in animals.The effects of glycerol monolaurate(GML)on intestinal innate immunity and associated molecular mechanisms were investigated using a chicken embryo model.Sixty-four Arbor Acres broiler embryos were randomly allocated into four groups.On embryonic day 17.5,the broiler embryos were administered with 9 mg of GML,which was followed by a 12-h incubation period and a12-h challenge with 32μg of lipopolysaccharide(LPS).On embryonic day 18.5,the jejunum and ileum were harvested.Results indicated that GML reversed the LPS-induced decline in villus height and upregulated the expression of mucin 2(P<0.05).GML decreased LPS-induced malondialdehyde production and boosted antioxidant enzyme activity(P<0.05).GML alleviated LPS-stimulated intestinal secretion of interleukin(IL)-1β,IL-6,and tumor necrosis factor-a(TNF-a)(P<0.05).GML also normalized LPS-induced changes in the gene expression of Toll-like receptor 4,nuclear factor kappa-B p65(NF-κB p65),cyclooxygenase-2,NOD-like receptor protein 3,IL-18,zonula occludens 1,and occludin(P<0.05).GML enhanced as well the expression of AMP-activated protein kinase a1 and claudin 1(P<0.05).In conclusion,GML improved intestinal morphology and antioxidant status by alleviating inflammatory responses and modulating NF-κB signaling in LPS-challenged broiler embryos.
基金supported by the National Natural Science Foundation of China,(No.82072217,81772135 and U21A20370)by the Jiangsu Natural Science Foundation(No.BK20201178).
文摘Background:Inflammation is an important factor in pathological scarring.The role of neutrophils,one of the most important inflammatory cells,in scar hyperplasia remains unclear.The purpose of this article is to study the correlation between neutrophil extracellular traps(NETs)and scar hyperplasia and identify a new target for inhibiting scar hyperplasia.Methods:Neutrophils were isolated from human peripheral blood by magnetic-bead sorting.NETs in plasma and scars were detected by enzyme-linked immunosorbent assays(ELISAs),immunofluorescence and flow cytometry.Immunohistochemistry was used to assess neutrophil(CD66B)infiltration in hypertrophic scars.To observe the entry of NETs into fibroblasts we used immunofluorescence and flow cytometry.Results:We found that peripheral blood neutrophils in patients with hypertrophic scars were more likely to form NETs(p<0.05).Hypertrophic scars showed greater infiltration with neutrophils and NETs(p<0.05).NETs activate fibroblasts in vitro to promote their differentiation and migration.Inhibition of NETs with cytochalasin in wounds reduced the hyperplasia of scars in mice.We induced neutrophils to generate NETs with different stimuli in vitro and detected the proteins carried by NETs.We did not find an increase in the expression of common scarring factors[interleukin(IL)-17 and transforming growth factor-β(TGF-β),p>0.05].However,inhibiting the production of NETs or degrading DNA reduced the differentiation of fibroblasts intomyofibroblasts.In vitro,NETs were found to be mediated by Toll-like receptor 9(TLR-9)in fibroblasts and further phosphorylated nuclear factor Kappa-B(NF-κB).We found that IL-6,which is downstream of NF-κB,was increased in fibroblasts.Additionally,IL-6 uses autocrine and paracrine signaling to promote differentiation and secretion.Conclusions:Our experiments found that NETs activate fibroblasts through the TLR-9/NF-κB/IL-6 pathway,thereby providing a new target for regulating hypertrophic scars.
文摘Heavy infection of the virus leads to overproduction of cytokines. The overproduction of cytokine (cytokines storms) is responsible for the critical cases and deaths of COVID-19. The nuclear factor kappa-B stimulates the expression of the genes, which is responsible for cytokines storm and RNA transcription. The COVID-19 virus can be controlled by inhibition of nuclear factor kappa-B. Nuclear factor kappa-B is controlled by inhibition of hydrogen peroxide and inhibitor kappa-B kinase enzyme.
基金supported by grants from the Project of Elitist Peak in Six Fields(No.2006-B-063)the Project of Medical Sciences(H200727),the Bureau of Health,Jiangsu Province,China
文摘BACKGROUND:The active form of nuclear factor-kappa B(NF- κB)is involved in the initiation,generation,and development of hepatocellular carcinoma(HCC),and is up-regulated in inflammation-associated malignancies.We investigated the dynamic expression of NF-κB and its influences on the occurrence of HCC through antiangiogenic(thalidomide) intervention in NF-κB activation. METHODS:Hepatoma models were induced with 2-fluorenyl- acetamide(2-FAA,0.05%)in male Sprague-Dawley rats,and thalidomide(100 mg/kg body weight)was administered intragastrically to intervene in NF-κB activation.The pathological changes in the liver of sacrificed rats were assessed after hematoxylin and eosin staining.NF-κB mRNA was amplified by RT-nested PCR.The alterations of NF-κB and vascular endothelial growth factor(VEGF)expression were analyzed by enzyme-linked immunosorbent assay,immunohistochemistry,and Western blotting. RESULTS:Rat hepatocytes showed denatured,precancerous,and cancerous stages in hepatocarcinogenesis,with an increasing tendency of hepatic NF-κB,NF-κB mRNA,and VEGF expression,and their values in the HCC group were higher than those in controls(P<0.001).In the thalidomide- treated group,the morphologic changes generated only punctiform denaturation and necrosis at the early or middle stages,and nodular hyperplasia or a little atypical hyperplasia at the final stages,with the expression of NF-κB (χ2=9.93,P<0.001)and VEGF(χ2=8.024,P<0.001)lower than that in the 2-FAA group. CONCLUSION:NF-κB is overexpressed in hepatocarcinogenesis and antiangiogenic treatment down-regulates the expression of NF-κB and VEGF,and delays the occurrence of HCC.
文摘BACKGROUND:Urinary trypsin inhibitor(UTI)inhibits the inflammatory response and protects against ischemia- reperfusion(I/R)injury.The inflammatory response is mediated by nuclear factor-kappa B(NF-κB)and its related target genes and products such as vascular endothelial cell adhesion molecule and CXC chemokines.We aimed to assess the roles of those mediators in a UTI-treated mouse model of hepatic I/R injury. METHODS:Treatment group 1(UTI given 5 minutes prior to liver ischemia),treatment group 2(UTI given 5 minutes after the anhepatic phase)and a control group were investigated.Blood and liver samples were obtained and compared at 1,3,6 and 24 hours after reperfusion. RESULTS:Attenuation of pathological hepatocellular damage was greater in the treatment groups than in the control group(P<0.05).Compared with the control group, the UTI treatment groups showed significantly lower serum alanine aminotransferase and aspartate aminotransferase levels,decreased myeloperoxidase activity,and reduced NF- κB activation.Also downregulated was the expression of tumor necrosis factor-alpha,cytokine-induced neutrophil chemoattractant,and macrophage inflammatory protein-2 at the mRNA level.P-selectin protein and intercellular adhesion molecule-1 protein expression were also downregulated.In addition,the treatment group 1 showed a better protective effect against I/R injury than the treatment group 2.CONCLUSIONS:UTI reduces NF-κB activation and downregulates the expression of its related mediators, followed by the inhibition of neutrophil aggregation and infiltration in hepatic I/R injury.The protective role of UTI is more effective in prevention than in treatment.
文摘AIM: To investigate whether NF-κB is activated in human gastric carcinoma tissues and, if so, to study whether there is any correlation between NF-κB activity and heparanase expression in gastric carcinoma.METHODS: NF-κB activation was assayed by immunohistochemical staining in formalin-fixed, paraffin-embedded specimens from 45 gastric carcinoma patients. Electrophoretic mobility shift assay (EMSA) method was used for nuclear protein from these fresh tissue specimens. Heparanase gene expression was quantified using quantitative RT-PCR.RESULTS: The nuclear translocation of RelA (marker of NF-κB activation) was significantly higher in tumor cells compared to adjacent and normal epithelial cells [(41.3±3.52)% vs (0.38±0.22) %, t= 10.993, P= 0.000＜0.05; (41.3±3.52)%vs (0±0.31)%, t = 11.484, P = 0.000＜0.05]. NF-κB activation was correlated with tumor invasion-related clinicopathological features such as lymphatic invasion,pathological stage, and depth of invasion (Z = 2.148,P= 0.032＜0.05; χ2 = 8.758, P= 0.033＜0.05; χ2 = 18.531,P = 0.006＜0.05). NF-κB activation was significantly correlated with expression of heparanase gene (r= 0.194,P= 0.046＜0.05).CONCLUSION: NF-κB RelA (p65) activation was related with increased heparanase gene expression and correlated with poor clinicopathological characteristics in gastric cancers. This suggests NF-κB as a major controller of the metastatic phenotype through its reciprocal regulation of some metastasis-related genes.
文摘Concomitantly with the increase in the prevalences of overweight/obesity, nonalcoholic fatty liver disease(NAFLD) has worldwide become the main cause of chronic liver disease in both adults and children. Patients with fatty liver display features of metabolic syndrome(Met S), like insulin resistance(IR), glucose intolerance, hypertension and dyslipidemia. Recently, epidemiological studies have linked obesity, Met S, and NAFLD to decreased bone mineral density and osteoporosis, highlighting an intricate interplay among bone, adipose tissue, and liver. Osteoprotegerin(OPG), an important symbol of the receptor activator of nuclear factor-B ligand/receptor activator of nuclear factor kappa B/OPG system activation, typically considered for its role in bone metabolism, may also play critical roles in the initiation and perpetuation of obesityrelated comorbidities. Clinical data have indicated that OPG concentrations are associated with hypertension, left ventricular hypertrophy, vascular calcification, endothelial dysfunction, and severity of liver damage in chronic hepatitis C. Nonetheless, the relationship between circulating OPG and IR as a key feature of Met S as well as between OPG and NAFLD remains uncertain. Thus, the aims of the present review are to provide the existent knowledge on these associations and to discuss briefly the underlying mechanisms linking OPG and NAFLD.
文摘AIM: To detect the nuclear factor kappa B (NF-κB) condition in human stage Ⅳ gastric carcinoma patients and to explore the correlation between NF- κB activation and survival of these patients after chemotherapy. METHODS: Expression of NF-κB-p65 was determined by immunohistochemical analysis. Activity of NF- κB DNA-binding in carcinoma tissue was detected by electrophoretic mobility shift assay. Kaplan-Meier survival analysis was performed to show the relation between NF-κB and progression-free survival (PFS) or overall survival (OS) of the patients. RESULTS: The positive expression rate of NF-κB-p65 in 60 gastric cancer tissue samples was 76.7% (46/60). The expression of NF-κB-p65 was reduced in adjacent carcinoma and normal tissue samples. Electrophoretic mobility shift assay (EMSA) analysis showed a strong activation of NF-κB in cancer tissue samples. A survival difference was found in NF- κB-p65 positive and negative patients. NF-κB-p65 expression was negative in cancer tissue samples (n = 14). PFS was 191.40 ± 59.88 d and 152.93 ± 16.99 d, respectively, in patients with positive NF- κB-p65 expression (n = 46) (P = 0.4028). Thesurvival time of patients with negative and positive NF-κB-p65 expression was 425.16 ± 61.61 d and 418.85 ± 42.98 d, respectively (P = 0.7303). Kaplan-Meier analysis showed no significant difference in PFS or OS. The 46 patient tissue which positive NF- κB-p65 expression was found in the tissue samples from the 46 patients whose PFS and OS were 564.89 ± 75.94 d and s 352.37 ± 41.32 d, respectively (P = 0.0165). CONCLUSION: NF-κB is activated in gastric carcinoma tissue, which is related to the OS after chemotherapy.
基金Supported by Scientific Research Foundation of the Chinese PLA Key Medical Programs During the 10th Five-Year Plan Period, No. 01Z011
文摘AIM: Nuclear factor kappa B (NF-κB) regulates a large number of genes involved in the inflammatory response to critical illnesses, but it is not known if and how NF-κB is activated and intercellular adhesion molecule-1 (ICAM-1)expressed in the gut following traumatic brain injury (TBI).The aim of current study was to investigate the temporal pattern of intestinal NF-κB activation and ICAM-1expression following TBI.METHODS: Male Wistar rats were randomly divided into six groups (6 rats in each group) including controls with sham operation and TBI groups at hours 3, 12, 24, and 72, and on d 7. Parietal brain contusion was adopted using weight-dropping method. All rats were decapitated at corresponding time point and mid-jejunum samples were taken. NF-κB binding activity in jejunal tissue was measured using EMSA. Immunohistochemistry was used for detection of ICAM-1 expression in jejunal samples.RESULTS: There was a very low NF-κB binding activity and little ICAM-1 expression in the gut of control rats after sham surgery. NF-κB binding activity in jejunum significantly increased by 160% at 3 h following TBI (P＜0.05 vs control), peaked at 72 h (500% increase)and remained elevated on d 7 post-injury by 390% increase. Compared to controls, ICAM-1 was significantly up-regulated on the endothelia of microvessels in villous interstitium and lamina propria by 24 h following TBI and maximally expressed at 72 h post-injury (P＜0.001). The endothelial ICAM-1 immunoreactivity in jejunal mucosa still remained strong on d 7 post-injury. The peak of NF-κB activation and endothelial ICAM-1 expression coincided in time with the period during which secondary mucosal injury of the gut was also at their culmination following TBI.CONCLUSION: TBI could induce an immediate and persistent up-regulation of NF-κB activity and subsequent up-regulation of ICAM-1 expression in the intestine.Inflammatory response mediated by increased NF-κB activation and ICAM-1 expression may play an important role in the pathogenesis of acute gut mucosal injury following TBI.
基金supported by grants from the Project of Elitist Peak in Six Fields(No.2006-B-063)the Projectof Medical Sciences(H200727),the Bureau of Health,Jiangsu Province,China
文摘BACKGROUND:Hepatocellular carcinoma(HCC)is one of the most common malignant tumors.We analyzed the expression of nuclear-transcription factor-kappa B(NF-κB) during hepatocarcinogenesis in order to evaluate its dynamic expression and its clinical value in the development and diagnosis of HCC. METHODS:Hepatoma models were induced by oral administration of 2-acetamidoflurene(2-FAA)to male Sprague-Dawley rats.Morphological changes were observed after hematoxylin and eosin staining.The cellular distribution of NF-κB expression during different stages of cancer development was investigated by immunohistochemistry, and the level of NF-κB expression in liver tissues was quantitatively analyzed by ELISA.The gene fragments of hepatic NF-κB were amplified by nested-polymerase chain reaction assay. RESULTS:Hepatocytes showed vacuole-like degeneration during the early stages,then had a hyperplastic nodal appearance during the middle stages,and finally progressed to tubercles of cancerous nests with high differentiation. The NF-κB-positive material was buff-colored,fine particles localized in the nucleus,and the incidence of NF-κB-positive cells was 81.8%in degeneration,83.3%in precancerous lesions,and 100%in cancerous tissues.All of these values were higher than those in controls(P<0.01). Hepatic NF-κB expression and hepatic NF-κB-mRNA were also higher during the course of HCC development(P<0.01).CONCLUSION:The NF-κB signal transduction pathway is activated during the early stages of HCC development, and its abnormal expression may be associated with the occurrence of HCC.
基金supported by the Leibniz Association,Germany,and the VELUX Foundation,Switzerland
文摘Activation of nuclear factor kappa B(NF-κB) is a hallmark of various central nervous system(CNS) pathologies. Neuron-specific inhibition of its transcriptional activator subunit RelA, also referred to as p65, promotes neuronal survival under a range of conditions, i.e., for ischemic or excitotoxic insults. In macro- and microglial cells, post-lesional activation of NF-κB triggers a growth-permissive program which contributes to neural tissue inflammation, scar formation, and the expression of axonal growth inhibitors. Intriguingly, inhibition of such inducible NF-κB in the neuro-glial compartment, i.e., by genetic ablation of RelA or overexpression of a transdominant negative mutant of its upstream regulator IκBα, significantly enhances functional recovery and promotes axonal regeneration in the mature CNS. By contrast, depletion of the NF-κB subunit p50, which lacks transcriptional activator function and acts as a transcriptional repressor on its own, causes precocious neuronal loss and exacerbates axonal degeneration in the lesioned brain. Collectively, the data imply that NF-κB orchestrates a multicellular program in which κB-dependent gene expression establishes a growth-repulsive terrain within the post-lesioned brain that limits structural regeneration of neuronal circuits. Considering these subunit-specific functions, interference with the NF-κB pathway might hold clinical potentials to improve functional restoration following traumatic CNS injury.
基金Supported by The National Natural Science Foundation of ChinaNO.81170374 and NO.81470842 to Hua J
文摘AIM: To investigate the effect of different dietary fatty acids on hepatic inflammasome activation.METHODS: Wild-type C57BL/6 mice were fed either a high-fat diet or polyunsaturated fatty acid(PUFA)-enriched diet. Primary hepatocytes were treated with either saturated fatty acids(SFAs) or PUFAs as well as combined with lipopolysaccharide(LPS). The expression of NOD-like receptor protein 3(NLRP3) inflammasome, peroxisome proliferator-activated receptor-γ and nuclear factor-kappa B(NF-κB) was determined by real-time PCR and Western blot. The activity of Caspase-1 and interleukine-1β production were measured.RESULTS: high-fat diet-induced hepatic steatosis was sufficient to induce and activate hepatic NLRP3 inflammasome. SFA palmitic acid(PA) directly activated NLRP3 inflammasome and increased sensitization to LPS-induced inflammasome activation in hepatocytes. In contrast, PUFA docosahexaenoic acid(Dh A) had thepotential to inhibit NLRP3 inflammasome expression in hepatocytes and partly abolished LPS-induced NLRP3 inflammasome activation. Furthermore, a highfat diet increased but PUFA-enriched diet decreased sensitization to LPS-induced hepatic NLRP3 inflammasome activation in vivo. Moreover, PA increased but Dh A decreased phosphorylated NF-κB p65 protein expression in hepatocytes.CONCLUSION:Hepatic NLRP 3 inflammasome activation played an important role in the development of non-alcoholic fatty liver disease. Dietary SFAs and PUFAs oppositely regulated the activity of NLRP3 inflammasome through direct activation or inhibition of NF-κB.
基金Supported by grants from National Natural Science Foundation of China, No. 30471614 (to DK Qiu) and No.30571730 (to X Ma)Shanghai Leading Academic Discipline Project, No.Y0205 (to X Ma)
文摘AIM: To investigate the role of p38 mitogen-activated protein kinase (p38MAPK) in murine experimental autoimmune hepatitis (EAH). METHODS: To induce EAH, the syngeneic S-100 antigen emulsified in complete Freud's adjuvant was injected intraperitoneally into adult male C57Bl/6 mice. Liver injury was assessed by serum ALT and liver histology. The expression and activity of p38 MAPK were measured by Western blot and kinase activity assays. In addition, DNA binding activities of nuclear factor kappa B (NF-κB) were analyzed by electrophoretic mobility shift assay. The effects of SB203580, a specifi c p38 MAPK inhibitor, on liver injuries and expression of proin? ammatory cytokines (interferon-γ, IL-12, IL-1β and TNF-α) were observed. RESULTS: The activity of p38 MAPK and NF-κB was increased and reached its peak 14 or 21 d after the fi rst syngeneic S-100 administration. Inhibition of p38 MAPK activation by SB203580 decreased the activation of NF-κB and the expression of proin? ammatory cytokines. Moreover, hepatic injuries were improved significantly after SB203580 administration. CONCLUSION: p38 MAPK and NF-κB play an important role in an animal model of autoimmune hepatitis (AIH) induced by autoantigens.
基金funded by the Health Research Fund from the Health Department of Shanxi Province, China, No.04015
文摘A mouse model of viral encephalitis was induced by intracranial injection of a Coxsackie virus B3 suspension. Quantitative real-time reverse transcription-PCR and western blot assay were applied to detect mRNA and protein expression of intelectin-2 and nuclear factor-kappa B in the viral encephalitis and control groups. Nuclear factor-kappa B and intelectin-2 mRNA and protein expression were significantly increased in mice with viral encephalitis. After intraperitoneal injection of Shuanghuanglian at a dose of 1.5 mg/kg for 5 successive days, intelectin-2 and nuclear factor-kappa B protein and mRNA expression were significantly decreased. To elucidate the relationship between intelectin-2 and nuclear factor-kappa B, mice with viral encephalitis were administered an intracerebral injection of 107 pfu recombinant lentivirus expressing intelectin shRNA. Both protein and mRNA levels of intelectin and nuclear factor-kappa B in brain tissue of mice were significantly decreased. Experimental findings suggest that Shuanghuanglian injection may downregulate nuclear factor-kappa B production via suppression of intelectin production, thus inhibiting inflammation associated with viral encephalitis.
基金Supported by The grant of the Romanian National Authority for Scientific Research,National Research Council-Executive Agency for Higher Education,Research,Development and Innovation Funding,No.PN-II-ID-PCE-2011-3-0591(grant awarded to Gafencu AV)the Romanian Academy,and the strategic grant financed by the European Social Found within the Sectorial Operational Program Human Resources Development 2007-2013,No.POSDRU/159/1.5/S/133391(Fenyo IM and Trusca VG)
文摘AIM: To investigate the effect of high homocysteine(Hcy) levels on apolipoprotein E(apoE) expression and the signaling pathways involved in this gene regulation.METHODS: Reverse transcriptase polymerase chain reaction(RT-PCR) and Western blot were used to assess apo E expression in cells treated with various concentrations(50-500 μmol/L) of Hcy. Calcium phosphatetransient transfections were performed in HEK-293 and RAW 264.7 cells to evaluate the effect of Hcy on apoE regulatory elements [promoter and distal multienhancer 2(ME2)]. To this aim, plasmids containing the proximal apoE promoter [(-500/+73)apoE construct] alone or in the presence of ME2 [ME2/(-500/+73)apoE construct] to drive the expression of the reporter luciferase gene were used. Co-transfection experiments were carried out to investigate the downstream effectors of Hcymediated regulation of apoE promoter by using specific inhibitors or a dominant negative form of IKβ. In other co-transfections, the luciferase reporter was under the control of synthetic promoters containing multiple specific binding sites for nuclear factor kappa B(NF-κB), activator protein-1(AP-1) or nuclear factor of activated T cells(NFAT). Chromatin immunoprecipitation(ChI P)assay was accomplished to detect the binding of NF-κB p65 subunit to the apoE promoter in HEK-293 treated with 500 μmol/L Hcy. As control, cells were incubated with similar concentration of cysteine. NF-κB p65 proteins bound to DNA were immunoprecipitated with anti-p65 antibodies and DNA was identified by PCR using primers amplifying the region-100/+4 of the apoE gene. RESULTS: RT-PCR revealed that high levels of Hcy(250-750 μmol/L) induced a 2-3 fold decrease in apoE m RNA levels in HEK-293 cells, while apo E gene expression was not significantly affected by treatment with lower concentrations of Hcy(100 μmol/L). Immunoblotting data provided additional evidence for the negative role of Hcy in apoE expression. Hcy decreased apoE promoter activity, in the presence or absence of ME2, in a dose dependent manner, in both RAW 264.7 and HEK-293 cells, as revealed by transient transfection experiments. The downstream effectors of the signaling pathways of Hcy were also investigated. The inhibitory effect of Hcy on the apo E promoter activity was counteracted by MAPK/ERK kinase 1/2(MEK1/2) inhibitor U0126, suggesting that MEK1/2 is involved in the downregulation of apoE promoter activity by Hcy. Our data demonstrated that Hcy-induced inhibition of apoE took place through activation of NF-κB. Moreover, we demonstrated that Hcy activated a synthetic promoter containing three NF-κB binding sites, but did not affect promoters containing AP-1 or NFAT binding sites. ChI P experiments revealed that NF-κB p65 subunit is recruited to the apoE promoter following Hcy treatment of cells.CONCLUSION: Hcy-induced stress negatively modulates apoE expression via MEK1/2 and NF-κB activation. The decreased apo E expression in peripheral tissues may aggravate atherosclerosis, neurodegenerative diseases and renal dysfunctions.
基金Supported by the Natural Science Foundation of Hu-Bei Province, No. 2002AB147
文摘AIM: To explore the effects of ketamine on hemo- dynamics, plasma proinflammatory cytokine (TNF-α and IL-6) levels and nuclear factor kappa B (NF-κB) activation during polymicrobial sepsis. METHODS: Male Sprague-Dawlay rats were subjected to cecal ligation and puncture (CLP) or sham operation. The rats were randomly assigned into four equal groups: sham CLP group, CLP group, ketamine (KT)Ⅰgroup and KTⅡgroup. Thirty minutes before CLP, ketamine (5 mg/kg per hour and 10 mg/kg per hour, respectively) was infused continuously through the left femoral vein cannula in KTⅠgroup or KTⅡgroup. Sham CLP group and CLP group received 0.9% saline only (5 mL/kg per hour). The right femoral artery was cannulated to monitor mean arterial pressure (MAP) and heart rates (HR),and draw blood samples. The proinflammatory cytokine (TNF-α and IL-6) levels of plasma were measured using enzyme-linked immunosorbent assays (ELISA). The hepatic NF-κB activation was determined by Western blot and HPIAS 2000 image analysis system. Twenty hours after CLP, the rats were killed by right femoral artery phlebotomization. RESULTS: CLP produced progressive hypotension, and a first increase followed by a decrease in HR. The hypotension was prevented, and the HR was slightly steady in ketamine treated rats. TNF-α levels of plasma reached a peak value at 2 h after CLP. Ketamine (KT Ⅰgroup or KTⅡgroup) caused a significant decrease compared with CLP group at 2, 5 and 9 h time points after CLP (14.3 ± 1.9 vs 4.3 ± 0.9, 9.7 ± 1.4 vs 4.3 ± 0.9; 9.3 ± 1.5 vs 4.3 ± 0.9, 8.7 ± 1.4 vs 4.3 ± 0.9; 6.0 ± 1.5 vs 5.0 ± 1.7, 5.3 ± 0.8 vs 5.0 ± 1.7; P < 0.01, respectively). The IL-6 levels of plasma firstly ascendedand then descended in CLP group, and reached a peak value at 9 h after CLP. Ketamine (KTⅠgroup or KTⅡ group) caused a significant decrease compared with CLP group at 5, 9 or 20 h after CLP (135.0 ± 52.6 vs 60.0 ± 16.3, 112.5 ± 52.6 vs 60.0 ± 16.3; 410.0 ± 68.7 vs 62.5 ± 12.5, 250.0 ± 28.0 vs 62.5 ± 12.5; 320.0 ± 25.9 vs 52.5 ± 10.1, 215.0 ± 44.6 vs 52.5 ± 10.1; P < 0.05, respectively). The IL-6 levels of plasma in KTⅡgroup were lower than those of KTⅠgroup at 9 h after CLP (250.0 ± 28.0 vs 410.0 ± 68.7; P < 0.05). In addition, CLP increased hepatic NF-κB expression compared with sham CLP. Ketamine suppressed NF-κB activation in a dose-dependent manner at 4 h after CLP (237.7 ± 3.5 vs 246.9 ± 3.1; P < 0.05). CONCLUSION: Ketamine stabilizes the hemodynamics, attenuates the proinflammatory cytokine responses, and inhibits hepatic NF-κB activation. These findings suggest that ketamine has protective effects against polymicrobial sepsis in rats.
基金Supported by The Project Research Fund from Kochi University,to Takeuchi Ha Grant-in-Aid for Scientific Research from the Ministry of Education,Science and Culture of Japan,No. 21590631 and 21590629,in part
文摘AIM:To investigate genetic diversity of Helicobacter pylori (H.pylori) cell division-related gene A (cdrA) and its effect on the host response.METHODS:Inactivation of H.pylori cdrA,which is involved in cell division and morphological elongation,has a role in chronic persistent infections.Genetic property of H.pylori cdrA was evaluated using polymerase chain reaction and sequencing in 128 (77 American and 51 Japanese) clinical isolates obtained from 48 and 51 patients,respectively.Enzyme-linked immunosorbent assay was performed to measure interleukin-8 (IL-8) secretion with gastric biopsy specimens obtained from American patients colonized with cdrA-positive or-negative strains and AGS cells cocultured with wild-type HPK5 (cdrA-positive) or its derivative HPKT510 (cdrA-disruptant).Furthermore,the cytotoxin-associated gene A (cagA) status (translocation and phosphorylation) and kinetics of transcription factors [nuclear factor-kappa B (NF-κB) and inhibition kappa B] were investigated in AGS cells co-cultured with HPK5,HPKT510 and its derivative HPK5CA (cagA disruptant) by western blotting analysis with immunoprecipitation.RESULTS:Genetic diversity of the H.pylori cdrA gene demonstrated that the cdrA status segregated into two categories including four allele types,cdrA-positive (allele types;Ⅰand Ⅱ) and cdrA-negative (allele types;Ⅲ and Ⅳ) categories,respectively.Almost all Japanese isolates were cdrA-positive (Ⅰ:7.8% and Ⅱ:90.2%),whereas 16.9% of American isolates were cdrA-positive (Ⅱ) and 83.1% were cdrA-negative (Ⅲ:37.7% and Ⅳ:45.5%),indicating extended diversity of cdrA in individual American isolates.Comparison of each isolate from different regions (antrum and corpus) in the stomach of 29 Americans revealed that cdrA status was identical in both isolates from different regions in 17 cases.However,12 cases had a different cdrA allele and 6 of them exhibited a different cdrA category between two regions in the stomach.Furthermore,in 5 of the 6 cases possessing a different cdrA category,cdrA-negative isolate existed in the corpus,suggesting that cdrA-negative strain is more adaptable to colonization in the corpus.IL-8 secretions from AGS revealed that IL-8 levels induced by a cdrA-disrupted HPKT510 was significantly lower (P < 0.01) compared to wildtype HPK5:corresponding to 50%-60% of those of wild-type HPK5.These data coincided with in vivo data that an average value of IL-8 in biopsy specimens from cdrA-positive and cdrA-negative groups was 215.6 and 135.9 pg/mL,respectively.Western blotting analysis documented that HPKT510 had no effect on CagA translocation and phosphorylation,however,nuclear accumulation of NF-κB was lower by HPKT510 compared to HPK5.CONCLUSION:Colonization by a cdrA-negative or cdrA-dysfunctional strain resulted in decreased IL-8 production and repression of NF-κB,and hence,attenuate the host immunity leading to persistent infection.
基金supported by the National Natural Science Foundation of China,No.81572191(to LC)and 81601067(to HZ)
文摘Progranulin is closely related to neuronal survival in a neuroinflammatory mouse model and attenuates inflammatory reactions. Atsttrin is an engineered protein composed of three progranulin fragments and has been shown to have an effect similar to that of progranulin. Atsttrin has anti-inflammatory actions in multiple arthritis mouse models, and it protects against further arthritis development. However, whether Atsttrin has a role in neuroinflammation remains to be elucidated. In this study, we produced a neuroinflammatory mouse model by intracerebroventricular injection of 1 μL lipopolysaccharide(10 μg/μL). Atsttrin(2.5 mg/kg) was administered via intraperitoneal injection every 3 days over a period of 7 days before intracerebroventricular injection of 1 μL lipopolysaccharide(10 μg/μL). In addition, astrocyte cultures were treated with 0, 100 or 300 ng/mL lipopolysaccharide, with 200 ng/mL Atsttrin simultaneously. Immunohistochemistry, enzyme-linked immunosorbent assay and real-time reverse transcription-polymerase chain reaction were performed to examine the protein and mRNA levels of inflammatory mediators and to assess activation of the nuclear factor kappa B signaling pathway. Progranulin expression in the brain of wild-type mice and in astrocyte cultures was increased after lipopolysaccharide administration. The protein and mRNA expression levels of tumor necrosis factor-α, interleukin-1β and inducible nitric oxide synthase were increased in the brain of progranulin knockout mice after lipopolysaccharide administration. Atsttrin treatment reduced the lipopolysaccharide-induced increase in the protein and mRNA levels of tumor necrosis factor-α, interleukin-1β, matrix metalloproteinase-3 and inducible nitric oxide synthase in the brain of progranulin knockout mice. Atsttrin also reduced the expression of cyclooxygenase-2, inducible nitric oxide synthase and matrix metalloproteinase 3 mRNA in lipopolysaccharide-treated astrocytes in vitro, and decreased the concentration of tumor necrosis factor α and interleukin-1β in the supernatant. Furthermore, Atsttrin significantly reduced the levels of phospho-nuclear factor kappa B inhibitor α in the brain of lipopolysaccharide-treated progranulin knockout mice and astrocytes, and it decreased the expression of nuclear factor kappa B2 in astrocytes. Collectively, our findings show that the anti-neuroinflammatory effect of Atsttrin involves inhibiton of the nuclear factor kappa B signaling pathway, and they suggest that Atsttrin may have clinical potential in neuroinflammatory therapy.
