摘要
目的:观察内质网应激相关分子CCAAT/增强子结合蛋白同源蛋白(CHOP)和Tribbles同源蛋白3(TRB3)在四氯化碳(CCl4)致大鼠肝纤维化过程中的表达变化,探讨其在肝纤维化过程中的可能作用。方法:体重180～200 g的雄性Wistar大鼠随机分为正常4周组、正常8周组、肝纤维化4周组和肝纤维化8周组,肝纤维化组大鼠皮下注射40%CCl4制备肝纤维化模型,分别在4周和8周处死大鼠,观察肝组织病理改变,Western blotting检测肝脏活化转录因子6(ATF6)蛋白,免疫组化、Western blotting和real-time PCR分别检测肝脏CHOP和TRB3蛋白和mRNA表达变化,TUNEL法检测肝脏细胞凋亡。结果:肝纤维化组大鼠肝脏可见假小叶形成,p90ATF6蛋白表达量较正常组明显减少(P<0.01),p50ATF6蛋白表达量较正常组明显增加(P<0.01),肝细胞胞浆CHOP和TRB3蛋白及mRNA表达量较正常组显著增加(P<0.01),细胞凋亡率较正常组显著增加(P<0.01)。结论:内质网应激相关分子CHOP和TRB3在CCl4致大鼠肝纤维化过程中蛋白及mRNA表达水平明显增加,其变化趋势与大鼠肝细胞凋亡率一致,提示内质网应激可能通过CHOP和TRB3促进肝细胞凋亡,参与肝纤维化发生发展。
AIM： To observe the changes of endoplasmic reticulum stress-related molecules CCAAT/enhancer- binding protein homologous protein（CHOP） and Tribbles homolog 3 （ TRB3 ） in the process of liver fibrosis induced by car- bon tetraehloride （ CC14 ）. METHODS ： Male Wistar rats weighing 180 g to 200 g were divided into 4-week normal control group, 8-week normal control group, 4-week liver fibrosis group and 8-week liver fibrosis group. The rats in liver fibrosis groups were induced by subcutaneous injection of 40% CC14 for 4 weeks or 8 weeks. The pathological changes of the liver were observed under light microscope. The protein level of ATF6 was determined by Western blotting. The protein and mR- NA levels of CHOP and TRB3 in the liver were analyzed by immunohistochemistry, Western blotting and real-time PCR, respectively. The apoptosis of hepatocytes was measured by TUNEL assay. RESULTS ： Pseudolobuli formed in the liver tissue of hepatic fibrotic rats. Compared with the control rats, the protein level of p90ATF6 was obviously decreased, the protein level of pSOATF6 was obviously increased, and the protein and mRNA levels of CHOP and TRB3 were obviously higher in the hepatocytes of hepatic fibrotic rats. The apoptosis of hepatocytes was also increased in 4-week and 8-week fi- brosis groups. CONCLUSION： In the process of liver fibrosis induced by 40% CC14, the obviously increased expression of endoplasmic retieulum stress-related molecules CHOP and TRB3 at protein and mRNA levels indicates that endoplasmic reticulum stress may play an important role in the liver fibrosis by promoting the apoptosis of hepatocytes.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2013年第5期906-912,共7页
Chinese Journal of Pathophysiology
基金
贵州省优秀科技教育人才省长专项资金项目[No.黔省专合字(2010)41号]
