FoxO1经ATF6调控Hcy诱导的肝细胞凋亡  

FoxO1 regulates Hcy-induced hepatocyte apoptosis through ATF6

作　　者：王青青 焦运[2] 吴欣妍 海秀玲 徐龙 张辉[1] 郭伟[1] 郝银菊[3] 姜怡邓[1,4,5] WANG Qing-qing;JIAO Yun;WU Xin-yan;HAI Xiu-ling;XU Long;ZHANG Hui;GUO Wei;HAO Yin-ju;JIANG Yi-deng(School of Basic Medicine,National Health Commission,Ningxia Medical University,Yinchuan 750004,China;Dept of Infectious Diseases,General Hospital of Ningxia Medical,National Health Commission,Ningxia Medical University,Yinchuan 750004,China;School of Pharmacy,NingxiaMedical University,National Health Commission,Ningxia Medical University,Yinchuan 750004,China;Ningxia Key Lab of Vascular Injury and Repair Research,National Health Commission,Ningxia Medical University,Yinchuan 750004,China;Key Lab of Metabolic Cardiovascular Disease Research,National Health Commission,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学基础医学院,宁夏银川750004 [2]宁夏医科大学总医院感染科,宁夏银川750004 [3]宁夏医科大学药学院,宁夏银川750004 [4]宁夏医科大学宁夏血管损伤与修复研究重点实验室,宁夏银川750004 [5]宁夏医科大学国家卫生健康委代谢性心血管疾病研究重点实验室,宁夏回族自治区,宁夏银川750004

出　　处：《中国药理学通报》2021年第2期203-209,共7页Chinese Pharmacological Bulletin

基　　金：宁夏自然科学基金资助项目(No 2018AAC03265,2019AAC03075);基础医学院“西部一流”建设项目(NXYLXK2017B07);临床医学一流学科开放课题(NXYLXK2017A05);宁夏回族自治区重点研发计划重大项目(No 2018BEG02004)。

摘　　要：目的探讨叉头状转录因子(FoxO1)在同型半胱氨酸(Hcy)诱导肝细胞凋亡中的作用及机制。方法随机选取10周龄雄性cbs^+/-与cbs+/+小鼠各8只饲以2%高蛋氨酸饮食12周,qRT-RCR和Western blot检测肝脏中FoxO1和ATF6的表达;100μmol·L^-1 Hcy处理肝细胞(HL-7702)48 h后,qRT-PCR和Western blot检测FoxO1和ATF6的表达改变;转染FoxO1和ATF6 siRNA后,Western blot检测ATF6、Bax和Bcl2蛋白表达,流式细胞术观察肝细胞凋亡情况;FoxO1过表达腺病毒与ATF6 siRNA共转至肝细胞,Western blot检测Bax和Bcl2蛋白表达。结果与cbs+/+组相比,cbs^+/-组肝脏中FoxO1和ATF6的表达水平显著增加(P<0.01);与Control组相比较,Hcy组FoxO1和ATF6的表达显著上调(P<0.01);与Hcy+siNC组相比,Hcy+siFoxO1组的ATF6和Bax表达水平明显降低,Bcl2的表达显著增加,肝细胞凋亡率显著降低(P<0.01);与Ad-FoxO1组相比,Ad-FoxO1+siATF6组Bax的表达显著降低,Bcl2的表达显著增加(P<0.01)。结论FoxO1介导的ATF6表达上调是Hcy致肝细胞凋亡进而引起肝损伤的重要机制之一。Aim To explore the role and possible mechanism of forked transcription factor(FoxO1)in hepatocyte apoptosis induced by homocysteine(Hcy).Methods The male cbs+/+and cbs^+/-mice of 10 weeks old were fed with 2%high methionine diet for 12 weeks,and then FoxO1 and ATF6 expressionswere detected by qRT-RCR and Western blot.FoxO1 and ATF6 expression in hepatocyte(HL-7702)treated with 100μmol·L^-1 Hcyfor 48 hours were detected by qRT-PCR and Western blot as well.FoxO1 siRNA was transfected in hepatocytes,the expressions of ATF6,Bax and Bcl-2 were detected by Western blot,and the apoptosis of hepatocyteswas observed by flow cytometry.FoxO1 overexpression adenovirus and ATF6 siRNA were co-transmitted to hepatocytes,and Bax and Bcl2 expression were detected by western blot.Results Compared with cbs+/+group,the expression of FoxO1 and ATF6 in liver of cbs^+/-group significantly increased(P<0.01).Compared with control group,the expression of FoxO1 and ATF6 in hepatocytes treated with Hcy was significantly up-regulated,while the expression of ATF6 and Bax in Hcy+siFoxO1 group was significantly lower than that in Hcy+siNC group,the expression of Bcl2 significantly increased,and the apoptotic rate of hepatocytes was significantly lower in Ad-FoxO1+siATF6 group than that in Ad-FoxO1 group(P<0.0l).Compared with Ad-FoxO1 group,the expression of Bax in Ad-FoxO1+siATF6 group was significantly lower and the expression of Bcl2 markedly higher than that in Ad-FoxO1 group.Conclusions The up-regulation of ATF6 mediated by FoxO1 may be one of the important mechanisms for Hcy-induced hepatocyte apoptosis and liver injury.

关 键 词：同型半胱氨酸 FOXO1 ATF6 内质网应激 肝细胞凋亡 肝损伤 

分 类 号：R-332[医药卫生—人体解剖和组织胚胎学] R322.47[医药卫生—基础医学]