茵陈蒿汤对阻塞性黄疸大鼠肝细胞ATF6/GRP78/CHOP凋亡信号通路的影响  

Influence of Yinchenhao decoction on the expression of ATF6/GRP78/CHOP apoptotic signaling pathway in hepatocyte of rats with obstructive jaundice

作　　者：孙一萌 刘浩 刘军舰 吴艳丽 张西波 戚经天 李忠廉 SUN Yi-meng;LIU Hao;LIU Jun-jian;WU Yan-li;Zhang Xi-bo;QI Jing-tian;LI Zhong-lian(Graduate School,Tianjin Medical University,Tianjin 300070,China;Department of Nephrology,Hubei Third People′s Hospital,Wuhan 430033,China;Department of Hepatobiliary and Pancreatic Surgery,Tianjin Hospital of Integrated Traditional Chinese and Western Medicine,Tianjin 300100,China;Department of Critical Care Medicine,Dongying Hospital of Traditional Chinese Medicine,Dongying 257055,China)

机构地区：[1]天津医科大学研究生院,天津300070 [2]湖北省第三人民医院肾内科,武汉430033 [3]天津市中西医结合医院肝胆胰外科二,天津300100 [4]东营市中医院重症医学科,东营257055

出　　处：《天津医科大学学报》2023年第3期252-257,共6页Journal of Tianjin Medical University

基　　金：国家自然科学基金面上项目(81273952);天津市中医药重点领域科技项目(2021006);天津市卫生健康委员会中医中西医结合科研课题(2021042);2022中央财政转移支付地方项目;天津市教委科研计划项目(2022KJ271)。

摘　　要：目的:探讨茵陈蒿汤对阻塞性黄疸大鼠肝细胞凋亡通路转录激活因子6(ATF6)、葡萄糖调节蛋白78(GRP78)、C/EBP同源蛋白(CHOP)及谷丙转氨酶(ALT)、天冬氨酸转氨酶(AST)的影响。方法:将正常大鼠肝细胞系(BRL-3A)分为胎牛血清配制的培养基培养的对照组(S组)、阻塞性黄疸血清配制的培养基培养的阻塞性黄疸组(O组)以及梗阻性黄疸血清+茵陈蒿汤含药血清配制的培养基培养的阻塞性黄疸+茵陈蒿汤组(Y组),分别于培养6、24、48 h后收集细胞。观察细胞形态且采用TUNEL法检测细胞凋亡情况,采用Western印迹和q-PCR法测定各组ATF6、GRP78及CHOP的蛋白及mRNA表达。采用ELISA法测定ALT、AST水平。结果:与S组相比,O组和Y组细胞各时间点均明显变小、变形和数量减少,凋亡指数明显升高,CHOP、ATF6和GRP78 mRNA及蛋白表达水平明显上调,上清液中AST、ALT含量均升高(F=29.75~154.3,均P<0.05);与O组相比较,Y组细胞状态在每个时间点均有显著改善,凋亡指数均下降,CHOP、ATF6和GRP78 mRNA及蛋白表达水平明显下调或趋于下调,上清液中AST、ALT含量均下降(F=29.75~154.3,均P<0.05)。结论:茵陈蒿汤可通过调控GRP78/ATF6/CHOP通路抑制细胞凋亡,减轻阻塞性黄疸引起的肝细胞损伤。Objective:To investigate the effects of Yinchenhao decoction on transcriptional activator 6(ATF6),glucoregulatory protein 78(GRP78),C/EBP homologous protein(CHOP),alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in in hepatocyte apoptosis pathway of obstructive jaundice rats.Methods:The normal rat hepatocytes line(BRL-3A)was divided into the control group(S group)of media culture prepared by fetal bovine serum,the obstructive jaundice group(O group)of media cultured by obstructive jaundice serum and obstructive jaundice+Yinchenhao decoction(Y group)prepared by medium culture prepared with obstructive jaundice serum+Yinchenhao decoction,and the cells were collected after 6 h,24 h and 48 h,respectively.The cell morphology was observed and apoptosis was detected by TUNEL method,and the protein and mRNA expression of ATF6,GRP78 and CHOP were determined by Western blotting and q-PCR.The levels of ALT and AST were determined by ELISA method.Results:Compared with the S group,the cells of the O group and Y group decreased significantly in size,deformation,and quantity at each time points,the apoptosis index increased significantly,the expression levels of ATF6,GRP78 and CHOP mRNA and protein were significantly regulated,and the contents of ALT and AST in the supernatant increased(F=154.3-29.75,all P<0.05).Compared with group O,the cell state of group Y was significantly improved at each time point,the apoptosis index decreased,the expression levels of CHOP,ATF6 and GRP78 mRNA and protein were significantly downregulated or tended to be down-regulated,and the contents of AST and ALT in the supernatant decreased(F=154.3-29.75,all P<0.05)。Conclusion:Yinchenhao decoction can inhibit apoptosis and reduce hepatocyte damage caused by obstructive jaundice by regulating ATF6/GRP78/CHOP pathway.

关 键 词：茵陈蒿汤 阻塞性黄疸 细胞凋亡 ATF6 GRP78 CHOP 

分 类 号：R575[医药卫生—消化系统]