摘要
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.
基金
Supported by the National Natural Science Foundation of China(No.82004251)
Sichuan Science and Technology Program(No.2022NSFSC1366)
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)。
