摘要
目的探讨Toll-样受体4(TLR4)信号通路阻断剂多粘菌素B(PMB)对脂多糖(LPS)诱导的大鼠肝星状细胞系(HSC-T6)自噬的影响并探讨自噬在肝纤维化(HF)中的意义。方法将常规培养的HSC-T6分为对照组、LPS组、PMB组、LPS+PMB组,各组细胞经相应处理后,以Western blot法检测各组微管相关蛋白轻链Ⅱ(LC3Ⅱ)及TLR4表达水平;免疫荧光法检测各组LC3在细胞内的分布情况;ELISA法检测各组细胞培养上清中层黏蛋白(LN)、透明质酸(HA)、Ⅲ型前胶原蛋白(PCⅢ)及Ⅳ型胶原蛋白(CⅣ)含量。结果LPS组HSC-T6中LC3含量、LC3Ⅱ蛋白表达水平及细胞培养上清中LN、HA、PCⅢ及CⅣ含量均高于对照组,差异有统计学意义(P<0.05);LPS+PMB组HSC-T6细胞中LC3含量、LC3Ⅱ蛋白表达水平及细胞培养上清中LN、HA、PCⅢ及CⅣ含量均低于LPS组,差异有统计学意义(P<0.05)。结论LPS可能通过活化TLR4信号通路进而促进HSC-T6细胞自噬及纤维化。
Objective To investigate the impacts of polymyxin B(PMB),the blocker of Toll-like receptor 4(TLR4)signaling pathway,on autophagy of rat hepatic stellate cell line(HSC-T6)induced by lipopolysaccharide(LPS)and the s ignificance of autophagy in hepatic fibros is(HF).Methods The conventionally cultured H SC-T6 were randomly divided into the control group,the LPS group,the PMB group and the LPS+PMB group.The expression levels of microtubule-associated protein light chainⅡ(LC3Ⅱ)and TLR4 in each group were detected by western blot method after corresponding treatment.Immunofluorescence method was used to detect the intracellular distribution of LC3 cells in each group.The contents of laminin(LN),hyaluronic acid(HA),procollagen typeⅢ(PCⅢ)and collagen typeⅣ(CⅣ)in cell culture supernatant were detected by ELISA method.Results The content of LC3 in HSC-T6,LC3Ⅱprotein expression level and the contents of LN,HA,PCⅢand CⅣin cell culture supernatant in the LPS group were all higher than those in the control group,with statistically significant differences(P<0.05).The contents of LC3,LC3Ⅱin HSC-T6 cells and LN,HA,PCⅢ,and CⅣin cell culture supernatant in the LPS+PMB group were all lower than those in the LPS group,with statistically significant differences(P<0.05).Conclusion LPS may further promote autophagy and fibrosis of HSC-T6 cells through activating TLR4 signaling path way.
作者
王美娇
徐军全
宋维芳
刘晓玲
WANG Meijiao;XU Junquan;SONG Weifang;LIU Xiaoling(Teaching and Research Section of Pathophysiology,Fenyang College of Shanxi Medical University,Shanxi,Fenyang 032200,China;Department of Gastroenterology,Fenyang Hospital in Shanxi Province,Shanxi,Fenyang 032200,China)
出处
《中国医药科学》
2022年第11期35-38,共4页
China Medicine And Pharmacy
基金
山西医科大学汾阳学院院级科研项目(2019D05)。
