AIM: To observe the effect of resveratrol on nuclear factor Kappa-B (NF-κB) activation and the inflammatory response in sodium taurocholate-induced pancreatitis in rats.METHODS: Seventy-two male SD rats were randomly...AIM: To observe the effect of resveratrol on nuclear factor Kappa-B (NF-κB) activation and the inflammatory response in sodium taurocholate-induced pancreatitis in rats.METHODS: Seventy-two male SD rats were randomly divided into three groups: sham operation group (control),severe acute pancreatitis (SAP) group, and severe acute pancreatitis group treated with resveratrol (RES). A SAP model was established by injecting 4% sodium taurocholate 1 mL/kg through puncturing the pancreatic duct. In Res group, Res was given at 30 mg/kg b.m. intraperitoneally after the SAP model was successfully established. Eight animals from each group were sacrificed at 3, 6 and 12 h after modeling. The expression of NF-κB activation of pancreas was detected by immunohistochemical staining, whereas the levels of TNF-α and IL-8 in pancreatic tissues were estimated by radioimmunoassay. The pathological changes of pancreas and lungs were examined microscopically.RESULTS: Much less hyperemia, edema, dust-colored necrotic focus and soaps were noticed in pancreas in RES group than in SAP group. In RES group, hemorrhage,exudates and infiltration of inflammatory cells in pancreas and interstitial edema, destruction of alveolar wall in lung were significantly less than in SAP group. In the SAP group,the activation of NF-κB in pancreatic tissues was enhanced significantly at any measure point compared with control group (64.23±10.72% vs2.56±0.65%, 55.86±11.34% vs 2.32±0.42%, 36.23±2.30% vs 2.40±0.36% ,P ＜0.01), TNF-α,IL-8 were also increased and reached their peak at 6 h and then declined. The activation of NF-κB and the levels of TNF-α and IL-8 in RES group were significantly lower than those in SAP group (P＜0.01): activation (52.63±9.45% vs 64.23±10.72%, 40.52±8.40% vs 55.86±11.34%, 29.83±5.37% vs36.23±2.30%), TN-α (132.76±15.68 pg/mL vs 158.36±12.58 pg/mL, 220.32±23.57 pg/mL vs 247.67± 11.62 pg/mL, 175.68±18.43 pg/mL vs 197.35±12.57 pg/mL) and IL-8 (0.62±0.21 μg/L vs0.83±0.10 μg/L, 1.10±0.124 μg/L vs1.32±0.18 μg/L, 0.98±0.16 μg/L vs 1.27±0.23 μg/L).CONCLUSION: The activation of NF-κB is involved in the inflammatory response of rats with SAP. Resveratrol could effectively inhibit the expression of NF-κB activation,alleviate the severity of SAP through its anti-inflammatory effects and regulate the inflammatory mediators.展开更多
Aim: To assess the spatial and temporal expression of germ cell nuclear factor (GCNF) in male mouse germ cells during postnatal development and in sperm before and after capacitation. Methods: The indirect immunofluor...Aim: To assess the spatial and temporal expression of germ cell nuclear factor (GCNF) in male mouse germ cells during postnatal development and in sperm before and after capacitation. Methods: The indirect immunofluorescence method with anti-GCNF antiserum was used to investigate the GCNF expression in mice at day 8, 10,14, 17, 20, 28, 35, 70, and 420 after birth and in sperm before and after capacitation. Results: With the proceeding of spermatogenesis, GCNF was first detected in the nuclei of spermatogonia and a few early stage primary spermatocytes at day 8, which was increased gradually at day 10 to 14 inclusive. From day 17 to day 20, the GCNF was concentrated in round spermatids, while both spermatogonia and early stage primary spermatocytes became GCNF negative. From day 28 until day 420, strong GCNF expression was shown in round spermatids and pachytene spermatocytes, while spermatogonia, early primary spermatocytes and elongating spermatids were all GCNF negative.In addition, it was also found that GCNF was localized on the acrosomal cap region of spermatozoa and there was a big change in GCNF expression during capacitation, from 98 % GCNF positive before capacitation to about 20 % positive following capacitation. The localization of GCNF in caput and cauda spermatozoa was similar. Conclusion:GCNF may play important roles in spermatogenesis, capacitation and fertilization. (Asian J Androl 2004 Sep; 6: 217-222)展开更多
AIM: Several epidemiological studies have demonstrated a dose association between Helicobacter pylori (H Pylon) infection and non-cardiac carcinoma of the stomach. H pylori infection induces active inflammation with n...AIM: Several epidemiological studies have demonstrated a dose association between Helicobacter pylori (H Pylon) infection and non-cardiac carcinoma of the stomach. H pylori infection induces active inflammation with neutrophilic infiltrations as well as production of oxygen free radicals that can cause DNA damage. The DNA damage induced by oxygen free radicals could have very harmful consequences,leading to gene modifications that are potentially mutagenic and/or carcinogenic. The aims of the present study were to assess the effect of H pyloriinfection on the expression of inducible nitric oxidative synthase (iNOS) and the production of 8-hydroxy-deoxyguanosine (8-OHdG), a sensitive marker of oxidative DNA injury in human gastric mucosa with and without tumor lesions, and to assess the possible factors affecting cell death signaling due tooxidative DNA damage.METHODS: In this study, 40 gastric carcinoma specimens and adjacent specimens were obtained from surgical resection. We determined the level of 8-OHdG formation by HPLC-ECD, and the expression of iNOS and mechanism of cell death signaling [including nuclear factor-κB(NFκB),MEKK-1, Caspase 3, B Cell lymphomal leukemia-2 (Bcl-2),inhibitor of apoptosis protein (IAP) and myeloid cellleukemia-1 (Mcl-1)] by Western-blot assay.RFSULTS: The concentrations of 8-OHdG, iNOS, NFx3, Mcl-1 and IAP were significantly higher in cancer tissues than in adjacent non-cancer tissues. In addition, significantly higher concentrations of 8-OHdG, iNOS, NFxB, Mcl-1 and lAP were detected in patients infected with H pyloricompared with patients who were not infected with HpylorL Furthermore,8-OHdG, iNOS, NFκB, Mcl-1 and IAP concentrations were significantly higher in stage 3 and 4 patients than in stage1 and 2 patients.CONCLUSION: Chronic Hpylori infection induces iNOS expression and subsequent DNA damage as well as enhances anti-apoptosis signal transduction. This sequence of events supports the rihypothesis that oxygen-free radical-mediated damage due to Hpyloriplays a pivotal role in the development of gastric carcinoma in patients with chronic gastritis.展开更多
AIM To study the molecular mechanisms involved in the hepatoprotective effects of naringenin(NAR)on carbon tetrachloride(CCl4)-induced liver fibrosis.METHODS Thirty-two male Wistar rats(120-150 g)were randomly divided...AIM To study the molecular mechanisms involved in the hepatoprotective effects of naringenin(NAR)on carbon tetrachloride(CCl4)-induced liver fibrosis.METHODS Thirty-two male Wistar rats(120-150 g)were randomly divided into four groups:(1)a control group(n=8)that received 0.7%carboxy methyl-cellulose(NAR vehicle)1 m L/daily p.o.;(2)a CCl4 group(n=8)that received 400 mg of CCl4/kg body weight i.p.3 times a week for 8 wk;(3)a CCl4+NAR(n=8)group that received 400 mg of CCl4/kg body weight i.p.3times a week for 8 wk and 100 mg of NAR/kg body weight daily for 8 wk p.o.;and(4)an NAR group(n=8)that received 100 mg of NAR/kg body weight daily for 8 wk p.o.After the experimental period,animals were sacrificed under ketamine and xylazine anesthesia.Liver damage markers such as alanine aminotransferase(ALT),alkaline phosphatase(AP),γ-glutamyl transpeptidase(γ-GTP),reduced glutathione(GSH),glycogen content,lipid peroxidation(LPO)and collagen content were measured.The enzymatic activity of glutathione peroxidase(GPx)was assessed.Liver histopathology was performed utilizing Masson’s trichrome and hematoxylin-eosin stains.Zymography assays for MMP-9 and MMP-2 were carried out.Hepatic TGF-β,α-SMA,CTGF,Col-I,MMP-13,NF-κB,IL-1,IL-10,Smad7,Smad3,p Smad3 and p JNK proteins were detected via western blot.RESULTS NAR administration prevented increases in ALT,AP,γ-GTP,and GPx enzymatic activity;depletion of GSH and glycogen;and increases in LPO and collagen produced by chronic CCl4 intoxication(P<0.05).Liver histopathology showed a decrease in collagen deposition when rats received NAR in addition to CCl4.Although zymography assays showed that CCl4 produced an increase in MMP-9 and MMP-2gelatinase activity;interestingly,NAR administration was associated with normal MMP-9 and MMP-2 activity(P<0.05).The anti-inflammatory,antinecrotic and antifibrotic effects of NAR may be attributed to its ability to prevent NF-κB activation and the subsequent production of IL-1 and IL-10(P<0.05).NAR completely prevented the increase in TGF-β,α-SMA,CTGF,Col-1,and MMP-13 proteins compared with the CCl4-treated group(P<0.05).NAR prevented Smad3phosphorylation in the linker region by JNK since this flavonoid blocked this kinase(P<0.05).CONCLUSION NAR prevents CCl4 induced liver inflammation,necrosis and fibrosis,due to its antioxidant capacity as a free radical inhibitor and by inhibiting the NF-κB,TGF-β-Smad3 and JNK-Smad3 pathways.展开更多
AIM: To observe the attenuation of ethanol extract of Herba Scutellaria barbata(SE) against diabetic retinopathy(DR) and its engaged mechanism.METHODS: C57BL/6J mice were intraperitoneally injected with streptozotocin...AIM: To observe the attenuation of ethanol extract of Herba Scutellaria barbata(SE) against diabetic retinopathy(DR) and its engaged mechanism.METHODS: C57BL/6J mice were intraperitoneally injected with streptozotocin(STZ, 55 mg/kg) for 5 consecutive days to induce diabetes. The diabetic mice were orally given with SE(100, 200 mg/kg) for 1mo at 1mo after STZ injection. Blood-retinal barrier(BRB) breakdown was detected by using Evans blue permeation assay. Real-time polymerase chain reaction(RT-PCR), Western blot and immunofluorescence staining were used to detect m RNA and protein expression. Enzyme-linked immunosorbent assay(ELISA) was used to detect serum contents of tumor necrosis factor-α(TNF-α) and interleukin(IL)-1β.RESULTS: SE(100, 200 mg/kg) reversed the breakdown of BRB in STZ-induced diabetic mice. The decreased expression of retinal claudin-1 and claudin-19, which are both tight junction(TJ) proteins, was reversed by SE. SE decreased the increased serum contents and retinal m RNA expression of TNF-α and IL-1β. SE also decreased the increased retinal expression of intercellular cell adhesion molecule-1(ICAM-1). SE reduced the increased phosphorylation of nuclear factor kappa B(NFκB) p65 and its subsequent nuclear translocation in retinas from STZinduced diabetic mice. Results of Western blot and retinal immunofluorescence staining of ionized calcium-binding adapter molecule 1(Iba1) demonstrated that SE abrogated the activation of microglia cells in STZ-induced diabetic mice.CONCLUSION: SE attenuates the development of DR by inhibiting retinal inflammation and restoring the decreased expression of TJ proteins including claudin-1 and claudin-19.展开更多
AIM To explore the effect of hydrogen sulfide(H2S)on restraint water-immersion stress(RWIS)-induced gastric lesions in rats and the influence of adenosine triphosphate(ATP)-sensitive potassium(KATP)channels and nuclea...AIM To explore the effect of hydrogen sulfide(H2S)on restraint water-immersion stress(RWIS)-induced gastric lesions in rats and the influence of adenosine triphosphate(ATP)-sensitive potassium(KATP)channels and nuclear factor kappa-light-chain-enhancer of activated B cells(NF-κB)pathway on such an effect.METHODS Male Wistar rats were randomly divided into a control group,a physiological saline(PS)group,a sodium hydrosulfide(Na HS)group,a glibenclamide(Gl)group,Gl plus Na HS group,a pyrrolidine dithiocarbamate(PDTC)group,and a PDTC plus Na HS group.Gastric mucosal injury was induced by RWIS for 3 h in rats,and gastric mucosal damage was analyzed after that.The PS,Na HS(100μmol/kg body weight),Gl(100μmol/kg body weight),Gl(100μmol/kg or 150μmol/kg body weight)plus Na HS(100μmol/kg body weight),PDTC(100μmol/kg body weight),and PDTC(100μmol/kg body weight)plus Na HS(100μmol/kg bodyweight)were respectively injected intravenously before RWIS.RESULTS RWIS induced serious gastric lesions in the rats in the PS pretreatment group.The pretreatment of Na HS(a H2S donor)significantly reduced the damage induced by RWIS.The gastric protective effect of the Na HS during RWIS was attenuated by PDTC,an NF-κB inhibitor,and also by glibenclamide,an ATP-sensitive potassium channel blocker,in a dose-dependent manner.CONCLUSION These results suggest that exogenous H2S plays a protective role against RWIS injury in rats,possibly through modulation of KATP channel opening and the NF-κB dependent pathway.展开更多
AIM To uncover the role of hepatocyte nuclear factor 4 alpha(HNF4α) in regulating hepatic expression of micro RNAs.METHODS Microarray and real-time PCR were used to determine hepatic expression of micro RNAs in young...AIM To uncover the role of hepatocyte nuclear factor 4 alpha(HNF4α) in regulating hepatic expression of micro RNAs.METHODS Microarray and real-time PCR were used to determine hepatic expression of micro RNAs in young-adult mice lacking Hnf4α expression in liver(Hnf4α-Liv KO). Integrative genomics viewer software was used to analyze the public chromatin immunoprecipitation-sequencing datasets for DNA-binding of HNF4α, RNA polymerase-Ⅱ, and histone modifications to loci of micro RNAs in mouse liver and human hepatoma cells. Dual-luciferase reporter assay was conducted to determine effects of HNF4α on the promoters of mouse and human micro RNAs as well as effects of micro RNAs on the untranslated regions(3'UTR) of two genes in human hepatoma cells. RESULTS Microarray data indicated that most micro RNAs remained unaltered by Hnf4α deficiency in Hnf4α-Liv KO mice. However, certain liver-predominant micro RNAs were down-regulated similarly in young-adult male and female Hnf4α-Liv KO mice. The down-regulation of mi R-101, mi R-192, mi R-193 a, mi R-194, mi R-215, mi R-802, and mi R-122 as well as induction of mi R-34 and mi R-29 in male Hnf4α-Liv KO mice were confirmed by real-timePCR. Analysis of public chromatin immunoprecipitationsequencing data indicates that HNF4α directly binds to the promoters of mi R-101, mi R-122, mi R-194-2/mi R-192 and mi R-193, which is associated with histone marks of active transcription. Luciferase reporter assay showed that HNF4α markedly activated the promoters of mouse and human mi R-101b/mi R-101-2 and the mi R-194/mi R-192 cluster. Additionally, mi R-192 and mi R-194 significantly decreased activities of luciferase reporters for the 3'UTR of histone H3F3 and chromodomain helicase DNA binding protein 1(CHD1), respectively, suggesting that mi R-192 and mi R-194 might be important in chromosome remodeling through directly targeting H3F3 and CHD1.CONCLUSION HNF4α is essential for hepatic basal expression of a group of liver-enriched micro RNAs, including mi R-101, mi R-192, mi R-193 a, mi R-194 and mi R-802, through which HNF4α may play a major role in the post-transcriptional regulation of gene expression and maintenance of the epigenome in liver.展开更多
AIM:To study the inhibition of nuclear factor kappa-B p65(NF-κB p65)antisense oligodeoxynucleotide(ASODN)on transdifferentiation of normal human lens epithelial cells induced by transforming growth factor-β2(TGF-β2...AIM:To study the inhibition of nuclear factor kappa-B p65(NF-κB p65)antisense oligodeoxynucleotide(ASODN)on transdifferentiation of normal human lens epithelial cells induced by transforming growth factor-β2(TGF-β2)in vitro.·METHODS:NF-κBp65ASODNand NF-κB p65 missense oligodeoxynucleotide(MSODN)were designed and synthesized.Human lens epithelial cell line(HLE B-3)cells were prepared for study and divided into 7 groups.Control group was HLE B-3 cells cultured in vitro in dulbecco's modified eagle medium(DMEM).T1,T2,and T3 group were HLE B-3 cells cultured in vitro in DMEM with 10 ng/m L TGF-β2 for 6h,12h,24h respectively.A+T group was HLE B-3 cells cultured with 10 ng/m L TGF-β2for 24h after transfected by NF-κB p65 ASODN for 24h.M+T group was HLE B-3 cells cultured with 10 ng/m L TGF-β2 for 24h after transfected by NF-κB p65 MSODN for 24h.The negative control group was HLE B-3 cells cultured with 10 ng/m L TGF-β2 for 24h after cultured with transfer agent(Hi Per Fect)for 24h.Cell morphology was observed at different time points using an inverted microscope.The expression of NF-κB p65 m RNA was detected with reverse transcription-polymerase chain reaction(RT-PCR),and the expression ofα-smooth muscle actin(α-SMA)protein was assayed with ELISA.·RESULTS:With the TGF-β2 stimulation prolongation,the expression of NF-κB p65 m RNA and a-SMA protein increased in T1,T2,T3 groups compared with the control group,and the difference was statistically significant(P<0.05).NF-κB p65 ASODN lowered the expression of NF-κB p65 m RNA andα-SMA protein induced by TGF-β2.NF-κB p65 MSODN and Hi Per Fect did not lower the expression of NF-κB p65 m RNA andα-SMA protein induced by TGF-β2.The difference between control group and A+T group was not statistically significant(P>0.05),but the difference among A+T group and other groups was statistically significant(P<0.05).·CONCLUSION:NF-κB p65 ASODN could lower the expression of NF-κB p65 m RNA andα-SMA protein induced by TGF-β2,and antagonized TGF-β2-induced transdifferentiation of HLE B-3 in vitro.NF-κB p65ASODN could be used as a new biological therapeutic target of posterior capsular opacification.展开更多
AIM To investigate whether single nucleotide polymorphisms in maf protein K(MAFK), which encodes the MAFK, lead to increased susceptibility to ulcerative colitis in the Japanese population. METHODS This case control s...AIM To investigate whether single nucleotide polymorphisms in maf protein K(MAFK), which encodes the MAFK, lead to increased susceptibility to ulcerative colitis in the Japanese population. METHODS This case control study examined the associations between MAFK single nucleotide polymorphisms(rs4268033 G>A, rs3735656 T>C and rs10226620 C>T) and ulcerative colitis susceptibility in 174 patients with ulcerative colitis(UC) cases, and 748 subjects without no lower abdominal symptoms, diarrhea or hematochezia(controls). In addition, as the second controls, we set 360 subjects, who have an irregular bowel movement without abnormal lower endoscopic findings(IBM controls).RESULTS The genotype frequency of rs4268033 AA and allelic frequency of the rs4268033 A allele were significantly higher in the UC cases than in both controls(P = 0.0005 and < 0.0001, P = 0.015 and 0.0027 vs controls and IBM controls, respectively). Logistic regression analysis after adjustment for age and gender showed that the rs4268033 AA and rs3735656 CC genotypes were significantly associated with susceptibility to UC development(OR = 2.63, 95%CI: 1.61-4.30, P = 0.0001 and OR = 1.81; 95%CI: 1.12-2.94, P = 0.015, respectively). Similar findings were observed by the comparison with IBM controls. In addition, the rs4268033 AA genotype was significantly associated with all phenotypes of UC except early onset. There was no significant association between rs10226620 and ulcerative colitis. CONCLUSION Our results provide the first evidence that MAFK genetic polymorphisms are significantly associated with susceptibility to UC development. In particular, rs4268033 is closely associated with an increased risk for the development of UC.展开更多
Objective: To explore the mechanism of the protective effects of Panax notoginseng saponins(PNS) on kidney in diabetic rats. Methods: Diabetic rat model was obtained by intravenous injection of alloxan, and the rats w...Objective: To explore the mechanism of the protective effects of Panax notoginseng saponins(PNS) on kidney in diabetic rats. Methods: Diabetic rat model was obtained by intravenous injection of alloxan, and the rats were divided into model, PNS-100 mg/(kg·day) and PNS-200 mg/(kg·day) groups, 10 each. Another 10 rats injected with saline were served as control. Periodic acid-Schiff staining and immunological histological chemistry were used to observe histomorphology and tissue expression of bone morphogenetic protein-7(BMP-7). Silent information regulator 1(SIRT1) was silenced in rat mesangial cells by RNA interference. The mR NA expressions of SIRT-1, monocyte chemoattractant protein-1(MCP-1), transforming growth factor β1(TGF-β1) and plasminogen activator inhibitor-1(PAI-1) were analyzed by reverse transcription polymerase chain reaction. The protein expressions of SIRT1 and the acetylation of nuclear factor κB(NF-κB) P65 were determined by western blotting. The concentration of MCP-1, TGF-β1 and malondialdehyde(MDA) in culture supernatant were detected by enzyme-linked immuno sorbent assay. The activity of superoxide dismutase(SOD) was detected by the classical method of nitrogen and blue four. Results: In diabetic model rats, PNS could not only reduce blood glucose and lipid(P<0.01), but also increase protein level of BMP-7 and inhibit PAI-1 expression for suppressing fibrosis of the kidney. In rat mesangial cells, PNS could up-regulate the expression of SIRT1(P<0.01) and in turn suppress the transcription of TGF-β1(P<0.05) and MCP-1(P<0.05). PNS could also reverse the increased acetylation of NF-κB p65 by high glucose. In addition, redox regulation factor MDA was down-regulated(P<0.05) and SOD was up-regulated(P<0.01), which were both induced by SIRT1 up-regulation. Conclusions: PNS could protect kidney from diabetes with the possible mechanism of up-regulating SIRT1, therefore inhibiting inflammation through decreasing the induction of inflammatory cytokines and TGF-β1, as well as activating antioxidant proteins.展开更多
Background This study was designed to investigate the potential pathogenicity of Mycoplasma penetrans (M. penetrans) and its molecular mechanisms responsible for the induction of iNOS gene expression in mouse macropha...Background This study was designed to investigate the potential pathogenicity of Mycoplasma penetrans (M. penetrans) and its molecular mechanisms responsible for the induction of iNOS gene expression in mouse macrophages stimulated by lipid-associated membrane proteins (LAMPs) prepared from M. penetrans.Methods Mouse macrophages were stimulated with M. penetrans LAMPs to assay the production of nitric oxide (NO). The expression of inducible nitric oxide synthase (iNOS) was detected by RT-PCR and Western blotting. The activity of nuclear factor κB (NF-κB) and the effects of pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-κB, on the production of nitric oxide and the expression of iNOS were also assessed in mouse macrophages treated with M. penetrans LAMPs by indirect immunofluorescence and Western blotting.Results M. penetrans LAMPs stimulated mouse macrophages to produce nitric oxide in a dose- and time-dependent manner. The mRNA and protein levels of iNOS were also upregulated in response to LAMP stimulation and inhibited by PDTC treatment. M. penetrans LAMPs were found to trigger NF-κB activation, a possible mechanism for the induction of iNOS expression.Conclusion This study demonstrated that M. penetrans may be an important etiological factor of certain diseases due to the ability of M. penetrans LAMPs to stimulate the expression of iNOS, which is probably mediated through the activation of NF-κB.展开更多
基金Supported by the National Natural Science Foundation of China, No.30371398
文摘AIM: To observe the effect of resveratrol on nuclear factor Kappa-B (NF-κB) activation and the inflammatory response in sodium taurocholate-induced pancreatitis in rats.METHODS: Seventy-two male SD rats were randomly divided into three groups: sham operation group (control),severe acute pancreatitis (SAP) group, and severe acute pancreatitis group treated with resveratrol (RES). A SAP model was established by injecting 4% sodium taurocholate 1 mL/kg through puncturing the pancreatic duct. In Res group, Res was given at 30 mg/kg b.m. intraperitoneally after the SAP model was successfully established. Eight animals from each group were sacrificed at 3, 6 and 12 h after modeling. The expression of NF-κB activation of pancreas was detected by immunohistochemical staining, whereas the levels of TNF-α and IL-8 in pancreatic tissues were estimated by radioimmunoassay. The pathological changes of pancreas and lungs were examined microscopically.RESULTS: Much less hyperemia, edema, dust-colored necrotic focus and soaps were noticed in pancreas in RES group than in SAP group. In RES group, hemorrhage,exudates and infiltration of inflammatory cells in pancreas and interstitial edema, destruction of alveolar wall in lung were significantly less than in SAP group. In the SAP group,the activation of NF-κB in pancreatic tissues was enhanced significantly at any measure point compared with control group (64.23±10.72% vs2.56±0.65%, 55.86±11.34% vs 2.32±0.42%, 36.23±2.30% vs 2.40±0.36% ,P ＜0.01), TNF-α,IL-8 were also increased and reached their peak at 6 h and then declined. The activation of NF-κB and the levels of TNF-α and IL-8 in RES group were significantly lower than those in SAP group (P＜0.01): activation (52.63±9.45% vs 64.23±10.72%, 40.52±8.40% vs 55.86±11.34%, 29.83±5.37% vs36.23±2.30%), TN-α (132.76±15.68 pg/mL vs 158.36±12.58 pg/mL, 220.32±23.57 pg/mL vs 247.67± 11.62 pg/mL, 175.68±18.43 pg/mL vs 197.35±12.57 pg/mL) and IL-8 (0.62±0.21 μg/L vs0.83±0.10 μg/L, 1.10±0.124 μg/L vs1.32±0.18 μg/L, 0.98±0.16 μg/L vs 1.27±0.23 μg/L).CONCLUSION: The activation of NF-κB is involved in the inflammatory response of rats with SAP. Resveratrol could effectively inhibit the expression of NF-κB activation,alleviate the severity of SAP through its anti-inflammatory effects and regulate the inflammatory mediators.
文摘Aim: To assess the spatial and temporal expression of germ cell nuclear factor (GCNF) in male mouse germ cells during postnatal development and in sperm before and after capacitation. Methods: The indirect immunofluorescence method with anti-GCNF antiserum was used to investigate the GCNF expression in mice at day 8, 10,14, 17, 20, 28, 35, 70, and 420 after birth and in sperm before and after capacitation. Results: With the proceeding of spermatogenesis, GCNF was first detected in the nuclei of spermatogonia and a few early stage primary spermatocytes at day 8, which was increased gradually at day 10 to 14 inclusive. From day 17 to day 20, the GCNF was concentrated in round spermatids, while both spermatogonia and early stage primary spermatocytes became GCNF negative. From day 28 until day 420, strong GCNF expression was shown in round spermatids and pachytene spermatocytes, while spermatogonia, early primary spermatocytes and elongating spermatids were all GCNF negative.In addition, it was also found that GCNF was localized on the acrosomal cap region of spermatozoa and there was a big change in GCNF expression during capacitation, from 98 % GCNF positive before capacitation to about 20 % positive following capacitation. The localization of GCNF in caput and cauda spermatozoa was similar. Conclusion:GCNF may play important roles in spermatogenesis, capacitation and fertilization. (Asian J Androl 2004 Sep; 6: 217-222)
文摘AIM: Several epidemiological studies have demonstrated a dose association between Helicobacter pylori (H Pylon) infection and non-cardiac carcinoma of the stomach. H pylori infection induces active inflammation with neutrophilic infiltrations as well as production of oxygen free radicals that can cause DNA damage. The DNA damage induced by oxygen free radicals could have very harmful consequences,leading to gene modifications that are potentially mutagenic and/or carcinogenic. The aims of the present study were to assess the effect of H pyloriinfection on the expression of inducible nitric oxidative synthase (iNOS) and the production of 8-hydroxy-deoxyguanosine (8-OHdG), a sensitive marker of oxidative DNA injury in human gastric mucosa with and without tumor lesions, and to assess the possible factors affecting cell death signaling due tooxidative DNA damage.METHODS: In this study, 40 gastric carcinoma specimens and adjacent specimens were obtained from surgical resection. We determined the level of 8-OHdG formation by HPLC-ECD, and the expression of iNOS and mechanism of cell death signaling [including nuclear factor-κB(NFκB),MEKK-1, Caspase 3, B Cell lymphomal leukemia-2 (Bcl-2),inhibitor of apoptosis protein (IAP) and myeloid cellleukemia-1 (Mcl-1)] by Western-blot assay.RFSULTS: The concentrations of 8-OHdG, iNOS, NFx3, Mcl-1 and IAP were significantly higher in cancer tissues than in adjacent non-cancer tissues. In addition, significantly higher concentrations of 8-OHdG, iNOS, NFxB, Mcl-1 and lAP were detected in patients infected with H pyloricompared with patients who were not infected with HpylorL Furthermore,8-OHdG, iNOS, NFκB, Mcl-1 and IAP concentrations were significantly higher in stage 3 and 4 patients than in stage1 and 2 patients.CONCLUSION: Chronic Hpylori infection induces iNOS expression and subsequent DNA damage as well as enhances anti-apoptosis signal transduction. This sequence of events supports the rihypothesis that oxygen-free radical-mediated damage due to Hpyloriplays a pivotal role in the development of gastric carcinoma in patients with chronic gastritis.
基金Supported by National Council of Science and Technology(Conacyt)of Mexico,No.253037 to Muriel P,and No.239516 to Segovia JFellowship No.358378 Hernández-Aquino E to from Conacytsupported by a grant of PRODEP(UABC-PTC-464)Mexico
文摘AIM To study the molecular mechanisms involved in the hepatoprotective effects of naringenin(NAR)on carbon tetrachloride(CCl4)-induced liver fibrosis.METHODS Thirty-two male Wistar rats(120-150 g)were randomly divided into four groups:(1)a control group(n=8)that received 0.7%carboxy methyl-cellulose(NAR vehicle)1 m L/daily p.o.;(2)a CCl4 group(n=8)that received 400 mg of CCl4/kg body weight i.p.3 times a week for 8 wk;(3)a CCl4+NAR(n=8)group that received 400 mg of CCl4/kg body weight i.p.3times a week for 8 wk and 100 mg of NAR/kg body weight daily for 8 wk p.o.;and(4)an NAR group(n=8)that received 100 mg of NAR/kg body weight daily for 8 wk p.o.After the experimental period,animals were sacrificed under ketamine and xylazine anesthesia.Liver damage markers such as alanine aminotransferase(ALT),alkaline phosphatase(AP),γ-glutamyl transpeptidase(γ-GTP),reduced glutathione(GSH),glycogen content,lipid peroxidation(LPO)and collagen content were measured.The enzymatic activity of glutathione peroxidase(GPx)was assessed.Liver histopathology was performed utilizing Masson’s trichrome and hematoxylin-eosin stains.Zymography assays for MMP-9 and MMP-2 were carried out.Hepatic TGF-β,α-SMA,CTGF,Col-I,MMP-13,NF-κB,IL-1,IL-10,Smad7,Smad3,p Smad3 and p JNK proteins were detected via western blot.RESULTS NAR administration prevented increases in ALT,AP,γ-GTP,and GPx enzymatic activity;depletion of GSH and glycogen;and increases in LPO and collagen produced by chronic CCl4 intoxication(P<0.05).Liver histopathology showed a decrease in collagen deposition when rats received NAR in addition to CCl4.Although zymography assays showed that CCl4 produced an increase in MMP-9 and MMP-2gelatinase activity;interestingly,NAR administration was associated with normal MMP-9 and MMP-2 activity(P<0.05).The anti-inflammatory,antinecrotic and antifibrotic effects of NAR may be attributed to its ability to prevent NF-κB activation and the subsequent production of IL-1 and IL-10(P<0.05).NAR completely prevented the increase in TGF-β,α-SMA,CTGF,Col-1,and MMP-13 proteins compared with the CCl4-treated group(P<0.05).NAR prevented Smad3phosphorylation in the linker region by JNK since this flavonoid blocked this kinase(P<0.05).CONCLUSION NAR prevents CCl4 induced liver inflammation,necrosis and fibrosis,due to its antioxidant capacity as a free radical inhibitor and by inhibiting the NF-κB,TGF-β-Smad3 and JNK-Smad3 pathways.
基金Supported by the National Natural Science Foundation of China(No.81173517No.81322053)
文摘AIM: To observe the attenuation of ethanol extract of Herba Scutellaria barbata(SE) against diabetic retinopathy(DR) and its engaged mechanism.METHODS: C57BL/6J mice were intraperitoneally injected with streptozotocin(STZ, 55 mg/kg) for 5 consecutive days to induce diabetes. The diabetic mice were orally given with SE(100, 200 mg/kg) for 1mo at 1mo after STZ injection. Blood-retinal barrier(BRB) breakdown was detected by using Evans blue permeation assay. Real-time polymerase chain reaction(RT-PCR), Western blot and immunofluorescence staining were used to detect m RNA and protein expression. Enzyme-linked immunosorbent assay(ELISA) was used to detect serum contents of tumor necrosis factor-α(TNF-α) and interleukin(IL)-1β.RESULTS: SE(100, 200 mg/kg) reversed the breakdown of BRB in STZ-induced diabetic mice. The decreased expression of retinal claudin-1 and claudin-19, which are both tight junction(TJ) proteins, was reversed by SE. SE decreased the increased serum contents and retinal m RNA expression of TNF-α and IL-1β. SE also decreased the increased retinal expression of intercellular cell adhesion molecule-1(ICAM-1). SE reduced the increased phosphorylation of nuclear factor kappa B(NFκB) p65 and its subsequent nuclear translocation in retinas from STZinduced diabetic mice. Results of Western blot and retinal immunofluorescence staining of ionized calcium-binding adapter molecule 1(Iba1) demonstrated that SE abrogated the activation of microglia cells in STZ-induced diabetic mice.CONCLUSION: SE attenuates the development of DR by inhibiting retinal inflammation and restoring the decreased expression of TJ proteins including claudin-1 and claudin-19.
基金Natural Science Foundation of Shandong Province,No.ZR2015CL016 and No.ZR2011CL012Colleges and Universities of Shandong Province Science and Technology Plan Projects,No.J11LC17
文摘AIM To explore the effect of hydrogen sulfide(H2S)on restraint water-immersion stress(RWIS)-induced gastric lesions in rats and the influence of adenosine triphosphate(ATP)-sensitive potassium(KATP)channels and nuclear factor kappa-light-chain-enhancer of activated B cells(NF-κB)pathway on such an effect.METHODS Male Wistar rats were randomly divided into a control group,a physiological saline(PS)group,a sodium hydrosulfide(Na HS)group,a glibenclamide(Gl)group,Gl plus Na HS group,a pyrrolidine dithiocarbamate(PDTC)group,and a PDTC plus Na HS group.Gastric mucosal injury was induced by RWIS for 3 h in rats,and gastric mucosal damage was analyzed after that.The PS,Na HS(100μmol/kg body weight),Gl(100μmol/kg body weight),Gl(100μmol/kg or 150μmol/kg body weight)plus Na HS(100μmol/kg body weight),PDTC(100μmol/kg body weight),and PDTC(100μmol/kg body weight)plus Na HS(100μmol/kg bodyweight)were respectively injected intravenously before RWIS.RESULTS RWIS induced serious gastric lesions in the rats in the PS pretreatment group.The pretreatment of Na HS(a H2S donor)significantly reduced the damage induced by RWIS.The gastric protective effect of the Na HS during RWIS was attenuated by PDTC,an NF-κB inhibitor,and also by glibenclamide,an ATP-sensitive potassium channel blocker,in a dose-dependent manner.CONCLUSION These results suggest that exogenous H2S plays a protective role against RWIS injury in rats,possibly through modulation of KATP channel opening and the NF-κB dependent pathway.
文摘AIM To uncover the role of hepatocyte nuclear factor 4 alpha(HNF4α) in regulating hepatic expression of micro RNAs.METHODS Microarray and real-time PCR were used to determine hepatic expression of micro RNAs in young-adult mice lacking Hnf4α expression in liver(Hnf4α-Liv KO). Integrative genomics viewer software was used to analyze the public chromatin immunoprecipitation-sequencing datasets for DNA-binding of HNF4α, RNA polymerase-Ⅱ, and histone modifications to loci of micro RNAs in mouse liver and human hepatoma cells. Dual-luciferase reporter assay was conducted to determine effects of HNF4α on the promoters of mouse and human micro RNAs as well as effects of micro RNAs on the untranslated regions(3'UTR) of two genes in human hepatoma cells. RESULTS Microarray data indicated that most micro RNAs remained unaltered by Hnf4α deficiency in Hnf4α-Liv KO mice. However, certain liver-predominant micro RNAs were down-regulated similarly in young-adult male and female Hnf4α-Liv KO mice. The down-regulation of mi R-101, mi R-192, mi R-193 a, mi R-194, mi R-215, mi R-802, and mi R-122 as well as induction of mi R-34 and mi R-29 in male Hnf4α-Liv KO mice were confirmed by real-timePCR. Analysis of public chromatin immunoprecipitationsequencing data indicates that HNF4α directly binds to the promoters of mi R-101, mi R-122, mi R-194-2/mi R-192 and mi R-193, which is associated with histone marks of active transcription. Luciferase reporter assay showed that HNF4α markedly activated the promoters of mouse and human mi R-101b/mi R-101-2 and the mi R-194/mi R-192 cluster. Additionally, mi R-192 and mi R-194 significantly decreased activities of luciferase reporters for the 3'UTR of histone H3F3 and chromodomain helicase DNA binding protein 1(CHD1), respectively, suggesting that mi R-192 and mi R-194 might be important in chromosome remodeling through directly targeting H3F3 and CHD1.CONCLUSION HNF4α is essential for hepatic basal expression of a group of liver-enriched micro RNAs, including mi R-101, mi R-192, mi R-193 a, mi R-194 and mi R-802, through which HNF4α may play a major role in the post-transcriptional regulation of gene expression and maintenance of the epigenome in liver.
基金Supported by the Outstanding Young Medical Personnel of Qingdao City
文摘AIM:To study the inhibition of nuclear factor kappa-B p65(NF-κB p65)antisense oligodeoxynucleotide(ASODN)on transdifferentiation of normal human lens epithelial cells induced by transforming growth factor-β2(TGF-β2)in vitro.·METHODS:NF-κBp65ASODNand NF-κB p65 missense oligodeoxynucleotide(MSODN)were designed and synthesized.Human lens epithelial cell line(HLE B-3)cells were prepared for study and divided into 7 groups.Control group was HLE B-3 cells cultured in vitro in dulbecco's modified eagle medium(DMEM).T1,T2,and T3 group were HLE B-3 cells cultured in vitro in DMEM with 10 ng/m L TGF-β2 for 6h,12h,24h respectively.A+T group was HLE B-3 cells cultured with 10 ng/m L TGF-β2for 24h after transfected by NF-κB p65 ASODN for 24h.M+T group was HLE B-3 cells cultured with 10 ng/m L TGF-β2 for 24h after transfected by NF-κB p65 MSODN for 24h.The negative control group was HLE B-3 cells cultured with 10 ng/m L TGF-β2 for 24h after cultured with transfer agent(Hi Per Fect)for 24h.Cell morphology was observed at different time points using an inverted microscope.The expression of NF-κB p65 m RNA was detected with reverse transcription-polymerase chain reaction(RT-PCR),and the expression ofα-smooth muscle actin(α-SMA)protein was assayed with ELISA.·RESULTS:With the TGF-β2 stimulation prolongation,the expression of NF-κB p65 m RNA and a-SMA protein increased in T1,T2,T3 groups compared with the control group,and the difference was statistically significant(P<0.05).NF-κB p65 ASODN lowered the expression of NF-κB p65 m RNA andα-SMA protein induced by TGF-β2.NF-κB p65 MSODN and Hi Per Fect did not lower the expression of NF-κB p65 m RNA andα-SMA protein induced by TGF-β2.The difference between control group and A+T group was not statistically significant(P>0.05),but the difference among A+T group and other groups was statistically significant(P<0.05).·CONCLUSION:NF-κB p65 ASODN could lower the expression of NF-κB p65 m RNA andα-SMA protein induced by TGF-β2,and antagonized TGF-β2-induced transdifferentiation of HLE B-3 in vitro.NF-κB p65ASODN could be used as a new biological therapeutic target of posterior capsular opacification.
文摘AIM To investigate whether single nucleotide polymorphisms in maf protein K(MAFK), which encodes the MAFK, lead to increased susceptibility to ulcerative colitis in the Japanese population. METHODS This case control study examined the associations between MAFK single nucleotide polymorphisms(rs4268033 G>A, rs3735656 T>C and rs10226620 C>T) and ulcerative colitis susceptibility in 174 patients with ulcerative colitis(UC) cases, and 748 subjects without no lower abdominal symptoms, diarrhea or hematochezia(controls). In addition, as the second controls, we set 360 subjects, who have an irregular bowel movement without abnormal lower endoscopic findings(IBM controls).RESULTS The genotype frequency of rs4268033 AA and allelic frequency of the rs4268033 A allele were significantly higher in the UC cases than in both controls(P = 0.0005 and < 0.0001, P = 0.015 and 0.0027 vs controls and IBM controls, respectively). Logistic regression analysis after adjustment for age and gender showed that the rs4268033 AA and rs3735656 CC genotypes were significantly associated with susceptibility to UC development(OR = 2.63, 95%CI: 1.61-4.30, P = 0.0001 and OR = 1.81; 95%CI: 1.12-2.94, P = 0.015, respectively). Similar findings were observed by the comparison with IBM controls. In addition, the rs4268033 AA genotype was significantly associated with all phenotypes of UC except early onset. There was no significant association between rs10226620 and ulcerative colitis. CONCLUSION Our results provide the first evidence that MAFK genetic polymorphisms are significantly associated with susceptibility to UC development. In particular, rs4268033 is closely associated with an increased risk for the development of UC.
基金Supported by National Natural Science Foundation of China(No.81273615)Zhejiang Provincial Natural Science Fund(No.Y2110849)
文摘Objective: To explore the mechanism of the protective effects of Panax notoginseng saponins(PNS) on kidney in diabetic rats. Methods: Diabetic rat model was obtained by intravenous injection of alloxan, and the rats were divided into model, PNS-100 mg/(kg·day) and PNS-200 mg/(kg·day) groups, 10 each. Another 10 rats injected with saline were served as control. Periodic acid-Schiff staining and immunological histological chemistry were used to observe histomorphology and tissue expression of bone morphogenetic protein-7(BMP-7). Silent information regulator 1(SIRT1) was silenced in rat mesangial cells by RNA interference. The mR NA expressions of SIRT-1, monocyte chemoattractant protein-1(MCP-1), transforming growth factor β1(TGF-β1) and plasminogen activator inhibitor-1(PAI-1) were analyzed by reverse transcription polymerase chain reaction. The protein expressions of SIRT1 and the acetylation of nuclear factor κB(NF-κB) P65 were determined by western blotting. The concentration of MCP-1, TGF-β1 and malondialdehyde(MDA) in culture supernatant were detected by enzyme-linked immuno sorbent assay. The activity of superoxide dismutase(SOD) was detected by the classical method of nitrogen and blue four. Results: In diabetic model rats, PNS could not only reduce blood glucose and lipid(P<0.01), but also increase protein level of BMP-7 and inhibit PAI-1 expression for suppressing fibrosis of the kidney. In rat mesangial cells, PNS could up-regulate the expression of SIRT1(P<0.01) and in turn suppress the transcription of TGF-β1(P<0.05) and MCP-1(P<0.05). PNS could also reverse the increased acetylation of NF-κB p65 by high glucose. In addition, redox regulation factor MDA was down-regulated(P<0.05) and SOD was up-regulated(P<0.01), which were both induced by SIRT1 up-regulation. Conclusions: PNS could protect kidney from diabetes with the possible mechanism of up-regulating SIRT1, therefore inhibiting inflammation through decreasing the induction of inflammatory cytokines and TGF-β1, as well as activating antioxidant proteins.
基金ThisstudywassupportedbyafundfromtheHunanProviceNaturalScienceFoundation (No 0 2JJY2 0 2 5 )
文摘Background This study was designed to investigate the potential pathogenicity of Mycoplasma penetrans (M. penetrans) and its molecular mechanisms responsible for the induction of iNOS gene expression in mouse macrophages stimulated by lipid-associated membrane proteins (LAMPs) prepared from M. penetrans.Methods Mouse macrophages were stimulated with M. penetrans LAMPs to assay the production of nitric oxide (NO). The expression of inducible nitric oxide synthase (iNOS) was detected by RT-PCR and Western blotting. The activity of nuclear factor κB (NF-κB) and the effects of pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-κB, on the production of nitric oxide and the expression of iNOS were also assessed in mouse macrophages treated with M. penetrans LAMPs by indirect immunofluorescence and Western blotting.Results M. penetrans LAMPs stimulated mouse macrophages to produce nitric oxide in a dose- and time-dependent manner. The mRNA and protein levels of iNOS were also upregulated in response to LAMP stimulation and inhibited by PDTC treatment. M. penetrans LAMPs were found to trigger NF-κB activation, a possible mechanism for the induction of iNOS expression.Conclusion This study demonstrated that M. penetrans may be an important etiological factor of certain diseases due to the ability of M. penetrans LAMPs to stimulate the expression of iNOS, which is probably mediated through the activation of NF-κB.
