AIM： To investigate the potential role of nuclear factor kappa-B （NF-κB） activation on the reactive oxygen species in rat acute necrotizing pancreatitis （ANP） and to assess the effect of pyrrolidine dithiocarbam...AIM： To investigate the potential role of nuclear factor kappa-B （NF-κB） activation on the reactive oxygen species in rat acute necrotizing pancreatitis （ANP） and to assess the effect of pyrrolidine dithiocarbamate （PDTC, an inhibitor of NF-κB）.METHODS： Rat ANP model was established by retrograde injection of 5% sodium taurocholate into biliopancreatic duct. Rats were randomly assigned to three groups （10 rats each）： Control group, ANP group and PDTC group. At the 6^th of the model, the changes of the serum amylase,nitric oxide （NO）, malondialdehyde （MDA）, superoxide dismutase （SOD） and pancreatic morphological damage were observed. The expressions of inducible nitric oxide （iNOS） were observed by SP immunohistochemistry. And bhe expressions of NF-κB p65 subunit mRNA were observed by hybridization in situ.RESULTS： Serum amylase and NO level decreased significantly in ANP group as compared with PDTC administrated group [（7 170.40＋1 308.63） U/L vs（4 074.10＋1 719.78） U/L,P〈0.05], [（76.95±9.04） μmol/L vs （65.18±9.02） μmol/L,P〈0.05] respectively. MDA in both ANP and PDTC group rose significantly over that in control group [（9.88＋1.52）nmol/L, （8.60±1.41） nmol/L, vs （6.04：hl.78） nmol/L,P〈0.05], while there was no significant difference between them. SOD levels in both ANP and PDTC group underwent a significant decrease as compared with that in control[（3 214.59±297.74） NU/mL, （3 260.62±229.44） NU/mL,vs（3 977.80＋309.09） NU/mL, P〈0.05], but there was no significant difference between them. Though they were still higher bhan those in Control group, pancreas destruction was slighter in PDTC group, iNOS expression and NF-κB p65 subunit mRNA expression were lower in PDTC group as compared with ANP group.CONCLUSION： We conclude that correlation among NF-κB activation, serum amylase, reactive oxygen species level and tissue damage suggests a key role of NF-κB in the pathogenesis of ANP. Inhibition of NF-κB activatio展开更多
Objective:To explore the effect and mechanism of Dahuang Zhechong Pill(DHZCP)on liver fibrosis.Methods:Liver fibrosis cell model was induced by transforming growth factor-β(TGF-β)in hepatic stellate cells(HSC-T6).DH...Objective:To explore the effect and mechanism of Dahuang Zhechong Pill(DHZCP)on liver fibrosis.Methods:Liver fibrosis cell model was induced by transforming growth factor-β(TGF-β)in hepatic stellate cells(HSC-T6).DHZCP medicated serum(DMS)was prepared in rats.HSC-T6 cells were divided into the control(15%normal blank serum culture),TGF-β(15%normal blank serum+5 ng/mL TGF-β),DHZCP(15%DMS+5 ng/mL TGF-β),DHZCP+PDTC[15%DMS+4 mmol/L ammonium pyrrolidine dithiocarbamate(PDTC)+5 ng/mL TGF-β],and PDTC groups(4 mmol/L PDTC+5 ng/mL TGF-β).Cell activity was detected by cell counting kit 8 and levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,IL-6,aspartate aminotransferase(AST)and alanine aminotransferase(ALT)in the cell supernatant were determined by enzymelinked immunosorbnent assay.Western blot was used to measure the expressions of p38 mitogen-activated protein kinase/nuclear factor kappa B/transforming growth factor-β1(p38 MAPK/NF-κB/TGF-β1)pathway related proteins,and the localization and expressions of these proteins were observed by immunofluorescence staining.Results:DHZCP improved the viability of cells damaged by TGF-βand reduced inflammatory cytokines and ALT and AST levels in the supernatant of HSC-T6 cells induced with TGF-β(P<0.05 or P<0.01).Compared with the TGF-βgroup,NF-κB p65 levels in the DHZCP group were decreased(P<0.05).p38 MAPK and NF-κB p65 levels in the DHZCP+PDTC were also reduced(P<O.01).Compared with the TGF-βgroup,the protein expression of Smad2 showed a downward trend in the DHZCP,DHZCP+PDTC,and PDTC groups(all P<0.01),and the decreasing trend of Samd3 was statistically significant only in DHZCP+PDTC group(P<0.01),whereas Smad7 was increased(P<0.05 or P<0.01).Conclusion:DHZCP can inhibit the process of HSC-T6 cell fibrosis by down-regulating the expression of p38 MAPK/NF-κB/TGF-β1 pathway.展开更多
文摘AIM： To investigate the potential role of nuclear factor kappa-B （NF-κB） activation on the reactive oxygen species in rat acute necrotizing pancreatitis （ANP） and to assess the effect of pyrrolidine dithiocarbamate （PDTC, an inhibitor of NF-κB）.METHODS： Rat ANP model was established by retrograde injection of 5% sodium taurocholate into biliopancreatic duct. Rats were randomly assigned to three groups （10 rats each）： Control group, ANP group and PDTC group. At the 6^th of the model, the changes of the serum amylase,nitric oxide （NO）, malondialdehyde （MDA）, superoxide dismutase （SOD） and pancreatic morphological damage were observed. The expressions of inducible nitric oxide （iNOS） were observed by SP immunohistochemistry. And bhe expressions of NF-κB p65 subunit mRNA were observed by hybridization in situ.RESULTS： Serum amylase and NO level decreased significantly in ANP group as compared with PDTC administrated group [（7 170.40＋1 308.63） U/L vs（4 074.10＋1 719.78） U/L,P〈0.05], [（76.95±9.04） μmol/L vs （65.18±9.02） μmol/L,P〈0.05] respectively. MDA in both ANP and PDTC group rose significantly over that in control group [（9.88＋1.52）nmol/L, （8.60±1.41） nmol/L, vs （6.04：hl.78） nmol/L,P〈0.05], while there was no significant difference between them. SOD levels in both ANP and PDTC group underwent a significant decrease as compared with that in control[（3 214.59±297.74） NU/mL, （3 260.62±229.44） NU/mL,vs（3 977.80＋309.09） NU/mL, P〈0.05], but there was no significant difference between them. Though they were still higher bhan those in Control group, pancreas destruction was slighter in PDTC group, iNOS expression and NF-κB p65 subunit mRNA expression were lower in PDTC group as compared with ANP group.CONCLUSION： We conclude that correlation among NF-κB activation, serum amylase, reactive oxygen species level and tissue damage suggests a key role of NF-κB in the pathogenesis of ANP. Inhibition of NF-κB activatio
基金Supported by the National Natural Science Foundation of China(No.82004251)Sichuan Science and Technology Program(No.2022NSFSC1366)+2 种基金China Postdoctoral Science Foundation(No.2022MD723716)the"Xinglin Scholars"Disciplinary Talent Research Improvement Plan–Youth Fund Talent Program(No.QJRC2022047)the Post Doctoral Program of Xinglin Scholar Discipline Talent Scientific Research Promotion Program of Chengdu University of Traditional Chinese Medicine(No.BSH2021029)。
文摘Objective:To explore the effect and mechanism of Dahuang Zhechong Pill(DHZCP)on liver fibrosis.Methods:Liver fibrosis cell model was induced by transforming growth factor-β(TGF-β)in hepatic stellate cells(HSC-T6).DHZCP medicated serum(DMS)was prepared in rats.HSC-T6 cells were divided into the control(15%normal blank serum culture),TGF-β(15%normal blank serum+5 ng/mL TGF-β),DHZCP(15%DMS+5 ng/mL TGF-β),DHZCP+PDTC[15%DMS+4 mmol/L ammonium pyrrolidine dithiocarbamate(PDTC)+5 ng/mL TGF-β],and PDTC groups(4 mmol/L PDTC+5 ng/mL TGF-β).Cell activity was detected by cell counting kit 8 and levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,IL-6,aspartate aminotransferase(AST)and alanine aminotransferase(ALT)in the cell supernatant were determined by enzymelinked immunosorbnent assay.Western blot was used to measure the expressions of p38 mitogen-activated protein kinase/nuclear factor kappa B/transforming growth factor-β1(p38 MAPK/NF-κB/TGF-β1)pathway related proteins,and the localization and expressions of these proteins were observed by immunofluorescence staining.Results:DHZCP improved the viability of cells damaged by TGF-βand reduced inflammatory cytokines and ALT and AST levels in the supernatant of HSC-T6 cells induced with TGF-β(P<0.05 or P<0.01).Compared with the TGF-βgroup,NF-κB p65 levels in the DHZCP group were decreased(P<0.05).p38 MAPK and NF-κB p65 levels in the DHZCP+PDTC were also reduced(P<O.01).Compared with the TGF-βgroup,the protein expression of Smad2 showed a downward trend in the DHZCP,DHZCP+PDTC,and PDTC groups(all P<0.01),and the decreasing trend of Samd3 was statistically significant only in DHZCP+PDTC group(P<0.01),whereas Smad7 was increased(P<0.05 or P<0.01).Conclusion:DHZCP can inhibit the process of HSC-T6 cell fibrosis by down-regulating the expression of p38 MAPK/NF-κB/TGF-β1 pathway.
