AIM:To determine the effect of ellagic acid on apop-tosis and proliferation in pancreatic cancer cells and to determine the mechanism of the pro-survival effects of ellagic acid. METHODS:The effect of ellagic acid on ...AIM:To determine the effect of ellagic acid on apop-tosis and proliferation in pancreatic cancer cells and to determine the mechanism of the pro-survival effects of ellagic acid. METHODS:The effect of ellagic acid on apoptosis was assessed by measuring Phosphatidylserine externalization,caspase activity,mitochondrial membrane potential and DNA fragmentation;and proliferation by measuring DNA thymidine incorporation. Mitochondrial membrane potential was measured in permeabilized cells,and in isolated mitochondria. Nuclear factor kB(NF-kB) activity was measured by electromobility shift assay(EMSA) . RESULTS:We show that ellagic acid,a polyphenolic compound in fruits and berries,at concentrations 10 to 50 mmol/L stimulates apoptosis in human pancreatic adenocarcinoma cells. Further,ellagic acid decreases proliferation by up to 20-fold at 50 mmol/L. Ellagic acid stimulates the mitochondrial pathway of apoptosis associated with mitochondrial depolarization,cytochrome C release,and the downstream caspaseactivation. Ellagic acid does not directly affect mitochondria. Ellagic acid dose-dependently decreased NF-kB binding activity. Furthermore,inhibition of NF-kB activity using IkB wild type plasmid prevented the effect of ellagic acid on apoptosis. CONCLUSION:Our data indicate that ellagic acid stimulates apoptosis through inhibition of the prosu-rvival transcription factor NF-kB.展开更多
AIM: To investigate the expression and role of nuclear factor κB(NF-κB) in diabetic retinopathy(DR) and its relationship with neovascularization and retinal cell apoptosis. METHODS: A total of 80 male Wistar rats we...AIM: To investigate the expression and role of nuclear factor κB(NF-κB) in diabetic retinopathy(DR) and its relationship with neovascularization and retinal cell apoptosis. METHODS: A total of 80 male Wistar rats were randomly assigned to control(4, 8, 12 and 16 wk, n =10 in each group) and diabetes mellitus(DM) groups(4, 8, 12 and 16wk, n =10 in each group). A diabetic rat model was established by intraperitoneal injection of streptozotocin(60 mg/kg). After 4, 8, 12 and 16 wk, rats were sacrificed.Retinal layers and retinal neovascularization growth were stained with hematoxylin-eosin and examined under light microscopy. Cell apoptosis in the retina was detected by Td T-mediated d UTP nick end labeling, and NF-κB distribution and expression in the retina was determined using immunohistochemistry. RESULTS: DM model success rate up to 100%.Diabetes model at each time point after the experimental groupcompared with the control group, the blood glucose was significantly increased, decreased body weight, each time point showed significant differences compared with the control group(P <0.01). After 12 wk other pathological changes in the retina of diabetic rats were observed; after 16 wk, neovascularization were observed. After 1mo, retinal cell apoptosis was observed.Compared with the control group, NF-κB expression in the DM group significantly increased with disease duration.CONCLUSION: With the prolonging of DM progression,the expression NF-κB increases. NF-κB may be related to retinal cell apoptosis and neovascularization.展开更多
The transcription factor nuclear factor κB(NF-κB) plays major roles in inflammatory diseases through regulation of inflammation and cell viability.Multiple sclerosis(MS) is a chronic inflammatory demyelinating and n...The transcription factor nuclear factor κB(NF-κB) plays major roles in inflammatory diseases through regulation of inflammation and cell viability.Multiple sclerosis(MS) is a chronic inflammatory demyelinating and neurodegenerative disease of the central nervous system(CNS).It has been shown that NF-κB is activated in multiple cell types in the CNS of MS patients,including T cells,microglia/macrophages,astrocytes,oligodendrocytes,and neurons.Interestingly,data from animal model studies,particularly studies of experimental autoimmune encephalomyelitis,have suggested that NF-κB activation in these individual cell types has distinct effects on the development of MS.In this review,we will cover the current literature on NF-κB and the evidence for its role in the development of MS and its animal model experimental autoimmune encephalomyelitis.展开更多
AIM: To investigate the role of nuclear factor κB(NF-κB) in the regulation of Epstein-Barr virus(EBV) latent membrane protein 1(LMP1) in EBV transformed cells. METHODS: LMP1 expression was examined in EBV transforme...AIM: To investigate the role of nuclear factor κB(NF-κB) in the regulation of Epstein-Barr virus(EBV) latent membrane protein 1(LMP1) in EBV transformed cells. METHODS: LMP1 expression was examined in EBV transformed human B lymphocytes with modulation of NF-κB activity. RESULTS: EBV infection is associated with several human cancers. EBV LMP1 is required for efficient transformation of adult primary B cells in vitro, and is expressed in several pathogenic stages of EBVassociated cancers. Regulation of EBV LMP1 involves both viral and cellular factors. LMP1 activates NF-κB signaling pathway that is a part of the EBV transformation program. However, the relation between NF-κB and LMP1 expression is not well established yet. In this report, we found that blocking the NF-κB activity by Inhibitor of κB stimulated LMP1 expression, while the overexpression of NF-κB repressed LMP1 expression in EBV-transformed IB4 cells. In addition, LMP1 repressed its own promoter activities in reporter assays, and the repression was associated with the activation of NF-κB. Moreover, NF-κB alone is sufficient to repress LMP1 promoter activities. CONCLUSION: Our data suggest LMP1 may repress its own expression through NF-κB in EBV transformed cells and shed a light on LMP1 regulation during EBV transformation.展开更多
背景:核转录因子κB信号通路在骨质疏松症的发病中扮演着重要的角色。近年来,大量的研究表明萜类中药单体化合物可以通过调控核转录因子κB信号通路,抑制骨吸收细胞的活性,促进骨形成细胞的分化,从而降低骨吸收并增加骨形成,对骨质疏松...背景:核转录因子κB信号通路在骨质疏松症的发病中扮演着重要的角色。近年来,大量的研究表明萜类中药单体化合物可以通过调控核转录因子κB信号通路,抑制骨吸收细胞的活性,促进骨形成细胞的分化,从而降低骨吸收并增加骨形成,对骨质疏松症具有一定的预防和治疗作用。目的:通过对国内外文献的分析和总结,深入研究核转录因子κB信号通路与骨质疏松症的关系,并对萜类中药单体化合物调控核转录因子κB信号通路防治骨质疏松症的作用机制进行阐明,同时对靶向调控核转录因子κB信号通路防治骨质疏松的萜类中药单体化合物进行系统性归纳。方法:由2名研究员根据拟定的纳入及排除标准以“NF-κB,骨质疏松症,成骨细胞,破骨细胞,血管生成,中药,萜类化合物”等为检索词检索中国知网数据库,以“NF-κB,osteoporosis,Osteoblasts,Osteoclasts,Angiogenesis,traditional Chinese medicine,terpenoid”等为检索词检索PubMed数据库相关文献,检索时间为建库至2022年12月,再通过第3名研究员对文献进行汇总和整理,最终纳入75篇文献进行系统性综述。结果与结论:①核转录因子κB信号通路能通过调控成骨细胞、破骨细胞的分化和增殖,以及血管生成,介导骨质疏松症的发病与进展。②核转录因子κB信号通路对成骨细胞的增殖和分化具有负调控的作用,激活核转录因子κB信号通路能增强破骨细胞的活性,抑制成骨细胞的生长,进而抑制代偿骨的生成保持骨稳态,但是过度激活核转录因子κB信号通路则会导致骨质疏松症。③核转录因子κB信号通路通过上调血管生成素1、血小板源性生长因子BB及血管内皮生长因子等细胞因子的表达水平,促进骨内血管生长,参与“血管生成-成骨”偶联。④萜类中药单体化合物在组织工程领域中具有促进骨细胞的增殖和分化,进而促进骨组织生长和修复的作用。⑤萜类中药单体化合物可以通过抑制核转录因子κB抑制蛋白降解,阻断核转录因子κB/P65蛋白磷酸化及核转位等过程,进而减弱核转录因子κB信号通路的传导,促进成骨细胞分化,抑制破骨细胞形成,起到防治骨质疏松的作用。⑥目前,萜类中药单体化合物调控核转录因子κB信号通路防治骨质疏松症的研究主要是基于体外细胞实验和动物模型,对于人体内复杂的生理和病理过程尚缺乏相关研究,未来需要开展更多的临床研究,进一步明确核转录因子κB信号通路参与干预骨质疏松症的作用机制和疗效。展开更多
AIM To inve s t igat e t he t he r ape ut ic e f f e c t of Jianpi Qingchang decoction(JPQCD) on dextran sulfate sodium(DSS)-induced ulcerative colitis(UC) in mice.METHODS C57BL/c mice were injected intragastrically w...AIM To inve s t igat e t he t he r ape ut ic e f f e c t of Jianpi Qingchang decoction(JPQCD) on dextran sulfate sodium(DSS)-induced ulcerative colitis(UC) in mice.METHODS C57BL/c mice were injected intragastrically with 5% DSS instead of drinking water for 7 d, and their body weight, diarrhea severity and fecal bleeding were monitored, while the mice in the control group were treated with standard drinking water, without DSS. After 7 d, the DSS drinking water was changed to normal water and the DSS group continued with DSS water. The control and DSS groups were given normal saline by intragastric injection. The 5-aminosalicylic acid(5-ASA) group was treated orally with 5-ASA at a dose of 100 mg/kg daily. The JPQCD group was treated orally with JPQCD at a dose of 17.1 g/kg daily. On day 14, the colon length was measured, the colorectalhistopathological damage score was assessed, and protein levels of interleukin(IL)-1β, IL-8 and tumor necrosis factor-alpha(TNF-α) in colon supernatants were measured by enzyme-linked immunosorbent assay. m RNA expression of IL-1β, IL-8, TNF-α and nuclear factor-kappa B(NF-κB) was detected by realtime quantitative polymerase chain reaction. Western blotting was used to detect the protein expression of NF-κB and inhibitor of kappa B. RESULTS Acute inflammation occurred in the mice administered DSS, including the symptoms of losing body weight, loose feces/watery diarrhea and presence of fecal blood; all these symptoms worsened at 7 d. The colons of mice treated with DSS were assessed by histological examination, and the results confirmed that acute inflammation had occurred, as evidenced by loss of colonic mucosa and chronic inflammatory cell infiltration, and these features extended into the deeper layer of the colon walls. The expression levels of IL-1β, IL-8 and TNF-α in the DSS group were higher than those in the control group(P < 0.05), and the expression levels of IL-1β, IL-8 and TNF-α in the JPQCD and 5-ASA groups were lower than those in the DSS group after treating with JPQCD and 5-ASA. Comparing with the DSS group, the mR NA level of IL-1β, IL-8, TNF-α and NF-κB was significantly reduced by 5-ASA and JPQCD. The difference between JPQCD and 5-ASA groups was not statistically significant(P > 0.05). Comparing with the DSS group, due to using JPQCD and 5-ASA, significant suppression of activation in DSSinduced NF-κB and increased phosphorylation of IκB in mice with experimental colitis occurred(P < 0.05). The difference between the JPQCD group and the 5-ASA group was not statistically significant(P > 0.05). CONCLUSION Activation of the NF-κB signaling pathway is inhibited by JPQCD, which shows the potential mechanism by which JPQCD treats UC.展开更多
AIM:To study effect of diterpenoid C extracted from radix curcumae on Helicobacter pylori(H.pylori)-infected inflammation,intestinal metaplasia,and nuclear factor kappa B(NF-κB)signaling pathway in vitro.METHODS:We u...AIM:To study effect of diterpenoid C extracted from radix curcumae on Helicobacter pylori(H.pylori)-infected inflammation,intestinal metaplasia,and nuclear factor kappa B(NF-κB)signaling pathway in vitro.METHODS:We used I-type H.pylori to infect human gastric epithelial gastric epithelium cell line(GES-1)cell lines,and then H.pylori-infected GES-1 cells were treated with radix curcumae(RC)-derived diterpenoid C of different concentrations(5,10,20μg/mL)and amoxicillin.The expression of p65,IκB kinase(IKK)αand IKKγproteins was detected with Western blotting,and the expression of interleukin(IL)-8,IL-6 and IL-4 was determined with enzyme-linked immunosorbent assay method.Data were analyzed using SPSS software ver18.0.For comparisons between groups of more than two unpaired values,one-way analysis of variance(ANOVA)was used.If an ANOVA F value was significant,post hoc comparisons were performed between groups.If results were not normally distributed,the Mann-Whitney U test was used to compare two groups of unpaired values,whereas for comparisons between groups of more than two unpaired values,the Kruskal-Wallis H test was used.Statistical significance was established at P<0.05.RESULTS:The MTT assay results revealed the inhibited rate of GES-1,and indicated that the IC5 of RCderived diterpenoid C and amoxicillin all were 5μg/mL for gastric GES-1 cells.The expression of IL-8 was significantly increased,especially at 12 h time point;and the expression of IL-4 was decreased in H.pyloriinfected GES-1 cells.After H.pylori-infected GES-1 cells were treated with RC-derived diterpenoid C of different concentrations and amoxicillin,the expression of IL-8was decreased at 12,24,48,72 h points(P<0.01),especially in high-concentration diterpenoid C(20μg/mL)group;and the expression of IL-4 was increased,especially in moderate and high-concentration diterpenoid C(10 and 20μg/mL)groups.RC-derived diterpenoid C had the inhibitory effects on H.pylori-induced p65 translocation from cytoplasm into cell nucleus,H.pylori-stimulant IkBαdegradation,the phosphorylation of p65 and IkBα,and the expression of IKKαand IKKβproteins.CONCLUSION:RC-derived diterpenoid C can block NF-κB signal pathway,effectively reducing the secretion of H.pylori-induced proinflammatory cytokine and increasing the secretion of anti-inflammatory cytokine.展开更多
AIM:To investigate the effect of propofol on human pancreatic cells and the molecular mechanism of propofol action.METHODS:We used the human pancreatic cancer cell line MIAPaCa-2 for in vitro studies measuring growth ...AIM:To investigate the effect of propofol on human pancreatic cells and the molecular mechanism of propofol action.METHODS:We used the human pancreatic cancer cell line MIAPaCa-2 for in vitro studies measuring growth inhibition and degree of apoptotic cell death induced by propofol alone,gemcitabine alone,or propofol followed by gemcitabine.All experiments were conducted in triplicate and carried out on three or more separate occasions.Data were means of the three or more independent experiments±SE.Statistically significant differences were determined by two-tailed unpaired Student’s t test and defined as P<0.05.RESULTS:Pretreatment of cells with propofol for 24 h followed by gemcitabine resulted in 24%-75% growth inhibition compared with 6%-18%when gemcitabine was used alone.Overall growth inhibition was directly correlated with apoptotic cell death.We also showed that propofol potentiated gemcitabine-induced killing by downregulation of nuclear factor-κB(NF-κB).In contrast,NF-κB was upregulated when pancreatic cancer cells were exposed to gemcitabine alone,suggesting a potential mechanism of acquired chemoresistance.CONCLUSION:Inactivation of the NF-κB signaling pathway by propofol might abrogate gemcitabineinduced activation of NF-κB,resulting in chemosensitization of pancreatic tumors to gemcitabine.展开更多
Background Nuclear factor κB (NF-κB) overactivation, requiring phosphorylation and degradation of its inhibitor IκBα, is the basis for chronicity of airway inflammation in asthma. Based on our previous plasmid pSh...Background Nuclear factor κB (NF-κB) overactivation, requiring phosphorylation and degradation of its inhibitor IκBα, is the basis for chronicity of airway inflammation in asthma. Based on our previous plasmid pShuttle-IκBα, carrying an IκBα gene from human placenta, we optimized a novel IκBα mutant (IκBαM) gene, constructed and characterized its replication-deficient recombinant adenovirus (AdIκBαM), and tested whether AdIκBαM-mediated overexpression of IκBαM could inhibit the NF-κB activation in endothelial cells. Methods IκBαM gene (203-1003 bp) encoding 267 amino acids, acquired by site-directed deleting N-terminal phosphorylation sites of serine 32/36, was subcloned into the pShuttle and pGEM-T vectors for further polymerase chain reaction (PCR), restriction digestion, deoxyribonucleic acid (DNA) sequencing and homology analyses. Subsequent to inserting the expression unit of pShuttle-IκBαM, containing cytomegalovirus (CMV) promoter, IκBαM complementary DNA (cDNA) and polyadenylic acid (PolyA) signals, into the type 5 adenovirus (Ad5) vector, the resultant AdIκBαM was packaged in human embryonic kidney (HEK) 293 cells by cotransfection with lipofectamine. Western blot analysis and electrophoretic mobility shift assay were utilized to detect the AdIκBαM-mediated overexpression of IκBαM in HEK293 cells and its suppressive effect on phorbol 12-myristate 13-acetate (PMA)-induced NF-κB activation in human umbilical vein endothelial (ECV304) cells, respectively. Results The relevant nucleotides and deduced amino acids of 801 bp IκBαM gene were consistent with those of IκBα gene (GenBank accession number: M69043). The titer of the prepared AdIκBαM was 4.0×1012 plaque-forming units (pfu)/L. Moreover, the IκBαM gene was overexpressed in HEK293 cells, and potently inhibited the PMA-induced NF-κB activation in ECV304 cells dose-dependently. Conclusions AdIκBαM is a novel vector for both efficient transfer and specific overexpression of IκBαM gene, as well as potent inhibition of NF-κB activity, providing a promising strategy for gene therapy of asthma.展开更多
AIM To explore the protective effects and underlying mechanisms of total polysaccharides of the Sijunzi decoction(TPSJ) on the epithelial barriers in vitro. METHODS Caco-2 cell monolayers were treated with or without ...AIM To explore the protective effects and underlying mechanisms of total polysaccharides of the Sijunzi decoction(TPSJ) on the epithelial barriers in vitro. METHODS Caco-2 cell monolayers were treated with or without TPSJ in the presence or absence of TNF-α, and paracellular permeability and transepithelial electrical resistance(TEER) were measured to evaluate the epithelial barrier function. Immunofluorescence and western blotting were respecti-vely used to evaluate the distribution and expression of the tight junction proteins claudin 1, claudin 2, zo3, and occludin in Caco-2 cells. western blotting was also used to evaluate the cellular expression of myosin light chain(MLC), phosphorylated MLC(pM LC), MLC kinase(MLCK), and nuclear factor(NF)-κB p65. RESULTS TPSJ promoted the proliferation of Caco-2 cells and inhibited TNF-α-induced secretion of pro-inflammatory cyto-kines. Furthermore, TPSJ significantly ameliorated both the reduction of TEER and the increased paracellular permeability observed in tumor necrosis factor(TNF)-α-damaged Caco-2 monolayers. Furthermore, TPSJ remarkably attenuated TNF-α-induced morphological changes, downregulated the expression of claudin 1, claudin 2, zo3, and occludin, and markedly suppressed TNF-α-mediated upregulation of p-MLC and MLCK expression. Finally, TPSJ inhibited the activation and expression of NF-κB p65. CONCLUSION Our results demonstrate that TPSJ alleviates the TNF-α-induced impairment of the intestinal epithelial cell barrier function by suppressing NF-κB p65-mediated phosphorylation of MLCK and MLC.展开更多
基金the Department of Veterans Affairs Merit Review (to A.S.G), the Hirshberg foundation and the NIH/NCCAM (1P01AT003960-01)
文摘AIM:To determine the effect of ellagic acid on apop-tosis and proliferation in pancreatic cancer cells and to determine the mechanism of the pro-survival effects of ellagic acid. METHODS:The effect of ellagic acid on apoptosis was assessed by measuring Phosphatidylserine externalization,caspase activity,mitochondrial membrane potential and DNA fragmentation;and proliferation by measuring DNA thymidine incorporation. Mitochondrial membrane potential was measured in permeabilized cells,and in isolated mitochondria. Nuclear factor kB(NF-kB) activity was measured by electromobility shift assay(EMSA) . RESULTS:We show that ellagic acid,a polyphenolic compound in fruits and berries,at concentrations 10 to 50 mmol/L stimulates apoptosis in human pancreatic adenocarcinoma cells. Further,ellagic acid decreases proliferation by up to 20-fold at 50 mmol/L. Ellagic acid stimulates the mitochondrial pathway of apoptosis associated with mitochondrial depolarization,cytochrome C release,and the downstream caspaseactivation. Ellagic acid does not directly affect mitochondria. Ellagic acid dose-dependently decreased NF-kB binding activity. Furthermore,inhibition of NF-kB activity using IkB wild type plasmid prevented the effect of ellagic acid on apoptosis. CONCLUSION:Our data indicate that ellagic acid stimulates apoptosis through inhibition of the prosu-rvival transcription factor NF-kB.
基金Supported by the National Natural Science Foundation of China (No.81371045) Shenyang Science and Technology Plan Project (No.F12-193-9-49)
文摘AIM: To investigate the expression and role of nuclear factor κB(NF-κB) in diabetic retinopathy(DR) and its relationship with neovascularization and retinal cell apoptosis. METHODS: A total of 80 male Wistar rats were randomly assigned to control(4, 8, 12 and 16 wk, n =10 in each group) and diabetes mellitus(DM) groups(4, 8, 12 and 16wk, n =10 in each group). A diabetic rat model was established by intraperitoneal injection of streptozotocin(60 mg/kg). After 4, 8, 12 and 16 wk, rats were sacrificed.Retinal layers and retinal neovascularization growth were stained with hematoxylin-eosin and examined under light microscopy. Cell apoptosis in the retina was detected by Td T-mediated d UTP nick end labeling, and NF-κB distribution and expression in the retina was determined using immunohistochemistry. RESULTS: DM model success rate up to 100%.Diabetes model at each time point after the experimental groupcompared with the control group, the blood glucose was significantly increased, decreased body weight, each time point showed significant differences compared with the control group(P <0.01). After 12 wk other pathological changes in the retina of diabetic rats were observed; after 16 wk, neovascularization were observed. After 1mo, retinal cell apoptosis was observed.Compared with the control group, NF-κB expression in the DM group significantly increased with disease duration.CONCLUSION: With the prolonging of DM progression,the expression NF-κB increases. NF-κB may be related to retinal cell apoptosis and neovascularization.
基金supported by grants from the National Institutes of Health(NS094151 and NS105689)the National Multiple Sclerosis Society(RG5239-A-3)(to WL)
文摘The transcription factor nuclear factor κB(NF-κB) plays major roles in inflammatory diseases through regulation of inflammation and cell viability.Multiple sclerosis(MS) is a chronic inflammatory demyelinating and neurodegenerative disease of the central nervous system(CNS).It has been shown that NF-κB is activated in multiple cell types in the CNS of MS patients,including T cells,microglia/macrophages,astrocytes,oligodendrocytes,and neurons.Interestingly,data from animal model studies,particularly studies of experimental autoimmune encephalomyelitis,have suggested that NF-κB activation in these individual cell types has distinct effects on the development of MS.In this review,we will cover the current literature on NF-κB and the evidence for its role in the development of MS and its animal model experimental autoimmune encephalomyelitis.
基金Supported by Grants from the NIH CA138213,RR15635Department of Defense W81XWH-12-1-0225(Luwen Zhang)Qianli Wang was partially supported by Undergraduate Creative Activities and Research Experiences and Beckman Scholars Program
文摘AIM: To investigate the role of nuclear factor κB(NF-κB) in the regulation of Epstein-Barr virus(EBV) latent membrane protein 1(LMP1) in EBV transformed cells. METHODS: LMP1 expression was examined in EBV transformed human B lymphocytes with modulation of NF-κB activity. RESULTS: EBV infection is associated with several human cancers. EBV LMP1 is required for efficient transformation of adult primary B cells in vitro, and is expressed in several pathogenic stages of EBVassociated cancers. Regulation of EBV LMP1 involves both viral and cellular factors. LMP1 activates NF-κB signaling pathway that is a part of the EBV transformation program. However, the relation between NF-κB and LMP1 expression is not well established yet. In this report, we found that blocking the NF-κB activity by Inhibitor of κB stimulated LMP1 expression, while the overexpression of NF-κB repressed LMP1 expression in EBV-transformed IB4 cells. In addition, LMP1 repressed its own promoter activities in reporter assays, and the repression was associated with the activation of NF-κB. Moreover, NF-κB alone is sufficient to repress LMP1 promoter activities. CONCLUSION: Our data suggest LMP1 may repress its own expression through NF-κB in EBV transformed cells and shed a light on LMP1 regulation during EBV transformation.
文摘背景:核转录因子κB信号通路在骨质疏松症的发病中扮演着重要的角色。近年来,大量的研究表明萜类中药单体化合物可以通过调控核转录因子κB信号通路,抑制骨吸收细胞的活性,促进骨形成细胞的分化,从而降低骨吸收并增加骨形成,对骨质疏松症具有一定的预防和治疗作用。目的:通过对国内外文献的分析和总结,深入研究核转录因子κB信号通路与骨质疏松症的关系,并对萜类中药单体化合物调控核转录因子κB信号通路防治骨质疏松症的作用机制进行阐明,同时对靶向调控核转录因子κB信号通路防治骨质疏松的萜类中药单体化合物进行系统性归纳。方法:由2名研究员根据拟定的纳入及排除标准以“NF-κB,骨质疏松症,成骨细胞,破骨细胞,血管生成,中药,萜类化合物”等为检索词检索中国知网数据库,以“NF-κB,osteoporosis,Osteoblasts,Osteoclasts,Angiogenesis,traditional Chinese medicine,terpenoid”等为检索词检索PubMed数据库相关文献,检索时间为建库至2022年12月,再通过第3名研究员对文献进行汇总和整理,最终纳入75篇文献进行系统性综述。结果与结论:①核转录因子κB信号通路能通过调控成骨细胞、破骨细胞的分化和增殖,以及血管生成,介导骨质疏松症的发病与进展。②核转录因子κB信号通路对成骨细胞的增殖和分化具有负调控的作用,激活核转录因子κB信号通路能增强破骨细胞的活性,抑制成骨细胞的生长,进而抑制代偿骨的生成保持骨稳态,但是过度激活核转录因子κB信号通路则会导致骨质疏松症。③核转录因子κB信号通路通过上调血管生成素1、血小板源性生长因子BB及血管内皮生长因子等细胞因子的表达水平,促进骨内血管生长,参与“血管生成-成骨”偶联。④萜类中药单体化合物在组织工程领域中具有促进骨细胞的增殖和分化,进而促进骨组织生长和修复的作用。⑤萜类中药单体化合物可以通过抑制核转录因子κB抑制蛋白降解,阻断核转录因子κB/P65蛋白磷酸化及核转位等过程,进而减弱核转录因子κB信号通路的传导,促进成骨细胞分化,抑制破骨细胞形成,起到防治骨质疏松的作用。⑥目前,萜类中药单体化合物调控核转录因子κB信号通路防治骨质疏松症的研究主要是基于体外细胞实验和动物模型,对于人体内复杂的生理和病理过程尚缺乏相关研究,未来需要开展更多的临床研究,进一步明确核转录因子κB信号通路参与干预骨质疏松症的作用机制和疗效。
基金Supported by the National Natural Science Foundation of China,No.81403355 and No.81573892the Project of 3-Year Action Plan for Shanghai Municipal Chinese Medicine Development,No.ZY3-RCPY-2-2001
文摘AIM To inve s t igat e t he t he r ape ut ic e f f e c t of Jianpi Qingchang decoction(JPQCD) on dextran sulfate sodium(DSS)-induced ulcerative colitis(UC) in mice.METHODS C57BL/c mice were injected intragastrically with 5% DSS instead of drinking water for 7 d, and their body weight, diarrhea severity and fecal bleeding were monitored, while the mice in the control group were treated with standard drinking water, without DSS. After 7 d, the DSS drinking water was changed to normal water and the DSS group continued with DSS water. The control and DSS groups were given normal saline by intragastric injection. The 5-aminosalicylic acid(5-ASA) group was treated orally with 5-ASA at a dose of 100 mg/kg daily. The JPQCD group was treated orally with JPQCD at a dose of 17.1 g/kg daily. On day 14, the colon length was measured, the colorectalhistopathological damage score was assessed, and protein levels of interleukin(IL)-1β, IL-8 and tumor necrosis factor-alpha(TNF-α) in colon supernatants were measured by enzyme-linked immunosorbent assay. m RNA expression of IL-1β, IL-8, TNF-α and nuclear factor-kappa B(NF-κB) was detected by realtime quantitative polymerase chain reaction. Western blotting was used to detect the protein expression of NF-κB and inhibitor of kappa B. RESULTS Acute inflammation occurred in the mice administered DSS, including the symptoms of losing body weight, loose feces/watery diarrhea and presence of fecal blood; all these symptoms worsened at 7 d. The colons of mice treated with DSS were assessed by histological examination, and the results confirmed that acute inflammation had occurred, as evidenced by loss of colonic mucosa and chronic inflammatory cell infiltration, and these features extended into the deeper layer of the colon walls. The expression levels of IL-1β, IL-8 and TNF-α in the DSS group were higher than those in the control group(P < 0.05), and the expression levels of IL-1β, IL-8 and TNF-α in the JPQCD and 5-ASA groups were lower than those in the DSS group after treating with JPQCD and 5-ASA. Comparing with the DSS group, the mR NA level of IL-1β, IL-8, TNF-α and NF-κB was significantly reduced by 5-ASA and JPQCD. The difference between JPQCD and 5-ASA groups was not statistically significant(P > 0.05). Comparing with the DSS group, due to using JPQCD and 5-ASA, significant suppression of activation in DSSinduced NF-κB and increased phosphorylation of IκB in mice with experimental colitis occurred(P < 0.05). The difference between the JPQCD group and the 5-ASA group was not statistically significant(P > 0.05). CONCLUSION Activation of the NF-κB signaling pathway is inhibited by JPQCD, which shows the potential mechanism by which JPQCD treats UC.
基金Supported by The Natural Science Foundation of Zhejiang Province of China,No.LY12H29002by grants of Scientific Research from Chinese Herbal Drug Administration,No.2011ZB032
文摘AIM:To study effect of diterpenoid C extracted from radix curcumae on Helicobacter pylori(H.pylori)-infected inflammation,intestinal metaplasia,and nuclear factor kappa B(NF-κB)signaling pathway in vitro.METHODS:We used I-type H.pylori to infect human gastric epithelial gastric epithelium cell line(GES-1)cell lines,and then H.pylori-infected GES-1 cells were treated with radix curcumae(RC)-derived diterpenoid C of different concentrations(5,10,20μg/mL)and amoxicillin.The expression of p65,IκB kinase(IKK)αand IKKγproteins was detected with Western blotting,and the expression of interleukin(IL)-8,IL-6 and IL-4 was determined with enzyme-linked immunosorbent assay method.Data were analyzed using SPSS software ver18.0.For comparisons between groups of more than two unpaired values,one-way analysis of variance(ANOVA)was used.If an ANOVA F value was significant,post hoc comparisons were performed between groups.If results were not normally distributed,the Mann-Whitney U test was used to compare two groups of unpaired values,whereas for comparisons between groups of more than two unpaired values,the Kruskal-Wallis H test was used.Statistical significance was established at P<0.05.RESULTS:The MTT assay results revealed the inhibited rate of GES-1,and indicated that the IC5 of RCderived diterpenoid C and amoxicillin all were 5μg/mL for gastric GES-1 cells.The expression of IL-8 was significantly increased,especially at 12 h time point;and the expression of IL-4 was decreased in H.pyloriinfected GES-1 cells.After H.pylori-infected GES-1 cells were treated with RC-derived diterpenoid C of different concentrations and amoxicillin,the expression of IL-8was decreased at 12,24,48,72 h points(P<0.01),especially in high-concentration diterpenoid C(20μg/mL)group;and the expression of IL-4 was increased,especially in moderate and high-concentration diterpenoid C(10 and 20μg/mL)groups.RC-derived diterpenoid C had the inhibitory effects on H.pylori-induced p65 translocation from cytoplasm into cell nucleus,H.pylori-stimulant IkBαdegradation,the phosphorylation of p65 and IkBα,and the expression of IKKαand IKKβproteins.CONCLUSION:RC-derived diterpenoid C can block NF-κB signal pathway,effectively reducing the secretion of H.pylori-induced proinflammatory cytokine and increasing the secretion of anti-inflammatory cytokine.
文摘AIM:To investigate the effect of propofol on human pancreatic cells and the molecular mechanism of propofol action.METHODS:We used the human pancreatic cancer cell line MIAPaCa-2 for in vitro studies measuring growth inhibition and degree of apoptotic cell death induced by propofol alone,gemcitabine alone,or propofol followed by gemcitabine.All experiments were conducted in triplicate and carried out on three or more separate occasions.Data were means of the three or more independent experiments±SE.Statistically significant differences were determined by two-tailed unpaired Student’s t test and defined as P<0.05.RESULTS:Pretreatment of cells with propofol for 24 h followed by gemcitabine resulted in 24%-75% growth inhibition compared with 6%-18%when gemcitabine was used alone.Overall growth inhibition was directly correlated with apoptotic cell death.We also showed that propofol potentiated gemcitabine-induced killing by downregulation of nuclear factor-κB(NF-κB).In contrast,NF-κB was upregulated when pancreatic cancer cells were exposed to gemcitabine alone,suggesting a potential mechanism of acquired chemoresistance.CONCLUSION:Inactivation of the NF-κB signaling pathway by propofol might abrogate gemcitabineinduced activation of NF-κB,resulting in chemosensitization of pancreatic tumors to gemcitabine.
基金This study was supported by grants from the National Youth NaturalScience Foundation of China(No.30400191)and the Key Subjectof’135’Project of Jiangsu Province(No.20013102).
文摘Background Nuclear factor κB (NF-κB) overactivation, requiring phosphorylation and degradation of its inhibitor IκBα, is the basis for chronicity of airway inflammation in asthma. Based on our previous plasmid pShuttle-IκBα, carrying an IκBα gene from human placenta, we optimized a novel IκBα mutant (IκBαM) gene, constructed and characterized its replication-deficient recombinant adenovirus (AdIκBαM), and tested whether AdIκBαM-mediated overexpression of IκBαM could inhibit the NF-κB activation in endothelial cells. Methods IκBαM gene (203-1003 bp) encoding 267 amino acids, acquired by site-directed deleting N-terminal phosphorylation sites of serine 32/36, was subcloned into the pShuttle and pGEM-T vectors for further polymerase chain reaction (PCR), restriction digestion, deoxyribonucleic acid (DNA) sequencing and homology analyses. Subsequent to inserting the expression unit of pShuttle-IκBαM, containing cytomegalovirus (CMV) promoter, IκBαM complementary DNA (cDNA) and polyadenylic acid (PolyA) signals, into the type 5 adenovirus (Ad5) vector, the resultant AdIκBαM was packaged in human embryonic kidney (HEK) 293 cells by cotransfection with lipofectamine. Western blot analysis and electrophoretic mobility shift assay were utilized to detect the AdIκBαM-mediated overexpression of IκBαM in HEK293 cells and its suppressive effect on phorbol 12-myristate 13-acetate (PMA)-induced NF-κB activation in human umbilical vein endothelial (ECV304) cells, respectively. Results The relevant nucleotides and deduced amino acids of 801 bp IκBαM gene were consistent with those of IκBα gene (GenBank accession number: M69043). The titer of the prepared AdIκBαM was 4.0×1012 plaque-forming units (pfu)/L. Moreover, the IκBαM gene was overexpressed in HEK293 cells, and potently inhibited the PMA-induced NF-κB activation in ECV304 cells dose-dependently. Conclusions AdIκBαM is a novel vector for both efficient transfer and specific overexpression of IκBαM gene, as well as potent inhibition of NF-κB activity, providing a promising strategy for gene therapy of asthma.
基金Supported by the National Natural Science Foundation of China,No.81202635the Guangdong Provincial Bureau of Chinese Medicine,No.20151244
文摘AIM To explore the protective effects and underlying mechanisms of total polysaccharides of the Sijunzi decoction(TPSJ) on the epithelial barriers in vitro. METHODS Caco-2 cell monolayers were treated with or without TPSJ in the presence or absence of TNF-α, and paracellular permeability and transepithelial electrical resistance(TEER) were measured to evaluate the epithelial barrier function. Immunofluorescence and western blotting were respecti-vely used to evaluate the distribution and expression of the tight junction proteins claudin 1, claudin 2, zo3, and occludin in Caco-2 cells. western blotting was also used to evaluate the cellular expression of myosin light chain(MLC), phosphorylated MLC(pM LC), MLC kinase(MLCK), and nuclear factor(NF)-κB p65. RESULTS TPSJ promoted the proliferation of Caco-2 cells and inhibited TNF-α-induced secretion of pro-inflammatory cyto-kines. Furthermore, TPSJ significantly ameliorated both the reduction of TEER and the increased paracellular permeability observed in tumor necrosis factor(TNF)-α-damaged Caco-2 monolayers. Furthermore, TPSJ remarkably attenuated TNF-α-induced morphological changes, downregulated the expression of claudin 1, claudin 2, zo3, and occludin, and markedly suppressed TNF-α-mediated upregulation of p-MLC and MLCK expression. Finally, TPSJ inhibited the activation and expression of NF-κB p65. CONCLUSION Our results demonstrate that TPSJ alleviates the TNF-α-induced impairment of the intestinal epithelial cell barrier function by suppressing NF-κB p65-mediated phosphorylation of MLCK and MLC.
