PSMD10在同型半胱氨酸激活肝细胞自噬促进肝脏损伤中的作用机制研究  

Mechanism of PSMD10 in homocysteine activating hepatocyte autophagy and promoting liver injury

作　　者：吴欣妍 焦运[4] 王青青 徐龙 张辉[1,2,3] 郭伟[1,2,3] 杨安宁 郝银菊[5] 姜怡邓[1,2,3] WU Xin-yan;JIAO Yun;WANG Qing-qing;XU Long;ZHANG Hui;GUO Wei;YANG An-ning;HAO Yin-ju;JIANG Yi-deng(School of Basic Medicine Ningxia Medical University,Ningxia Hui Autonomous Region,Yinchuan 750004,China;Key Lab of Metabolic Cardiovascular Disease Research,National Health Commission Ningxia Medical University,Ningxia Hui Autonomous Region,Yinchuan 750004,China;Ningxia Key Lab of Vascular Injury and Repair ResearchNingxia Medical University,Ningxia Hui Autonomous Region,Yinchuan 750004,China;Dept of Infection,Ningxia Medical University,Ningxia Hui Autonomous Region,Yinchuan 750004,China;School of Pharmacy,Ningxia Medical University,Ningxia Hui Autonomous Region,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学基础医学院,宁夏回族自治区,宁夏银川750004 [2]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 [3]宁夏医科大学宁夏血管损伤与修复研究重点实验室,宁夏回族自治区,宁夏银川750004 [4]宁夏医科大学总医院感染科,宁夏回族自治区,宁夏银川750004 [5]宁夏医科大学药学院,宁夏回族自治区,宁夏银川750004

出　　处：《中国药理学通报》2021年第4期535-541,共7页Chinese Pharmacological Bulletin

基　　金：宁夏回族自治区重点研发计划一般项目(No 2020BEG03005);宁夏回族自治区重点研发计划重点项目(No 2018BEG02004,2020BFH02003);国家自然科学基金地区项目(No 82060110)。

摘　　要：目的探讨26S蛋白酶体非ATP酶调节亚基10(PSMD10)在同型半胱氨酸激活肝细胞自噬促进肝脏损伤中的作用机制。方法选取cbs^(+/+)和cbs^(+/-)小鼠各8只,2%高蛋氨酸饮食饲养8周。全自动生化仪检测血清中AST和ALT水平;Western blot检测肝脏LC3B、p62和PSMD10蛋白表达;qRT-PCR检测PSMD10 mRNA表达;100μmol Hcy干预肝细胞(HL-7702);ELISA检测培养液上清中AST和ALT水平;Western blot检测LC3B、p62和PSMD10蛋白表达;qRT-PCR检测PSMD10 mRNA的表达;转染PSMD10 siRNA或过表达腺病毒,Western blot检测LC3B和p62的表达;ELISA检测AST和ALT水平。结果与cbs^(+/+)组比较,cbs^(+/-)组血清AST和ALT的水平增加(P<0.01),自噬水平增加(P<0.01),PSMD10表达水平增加(P<0.05);细胞水平与动物水平结果一致;与Hcy+siNC组相比,LC3Ⅱ/Ⅰ的比值降低(P<0.01),p62的蛋白表达增加(P<0.01),AST和ALT的浓度降低(P<0.01);与ad-GFP组相比,ad-PSMD10组LC3Ⅱ/Ⅰ比值增加(P<0.01),p62的蛋白表达降低(P<0.01),AST和ALT的浓度增加(P<0.01)。结论Hcy通过上调PSMD10的表达导致肝细胞过度自噬,造成肝脏损伤。Aim To discuss the mechanism of proteasome 26S subunit non-ATPase 10(PSMD10)activating hepatocyte autophagy and promoting liver injury by in homocysteine.Methods Wild mice and cystathionineβ-synthase(CBS)gene knockout mice were used and divided into normal(cbs^(+/+),n=8)group and gene knockout(cbs^(+/-),n=8)group,and the mice were fed with high methionine diet for eight weeks.Serum levels of AST and ALT were detected by automatic biochemical analyzer.The expression changes of autophagy-related proteins LC3B,p62 and PSMD10 in mouse liver were detected by Western blot.The expression of PSMD10 mRNA in mouse liver was detected by qRT-PCR.For cells,HL-7702 hepatocytes were cultured in vitro,divided into control group and Hcy group;AST and ALT levels were detected by ELISA.The expression changes of autophagy-related proteins LC3B,p62 and PSMD10 in hepatocytes were detected by Western blot.The expression of PSMD10 mRNA in hepatocytes were detected by qRT-PCR.Hepatocytes were transfected with PSMD10 siRNA,Hcy intervention for 48h or transfected PSMD10 overexpressed adenovirus,Western blot was used to detect the expression of autophagy-related proteins LC3B and p62,then AST and ALT levels were detected by ELISA.Results For animals,compared with cbs^(+/+)mice,the levels of AST and ALT in cbs^(+/-)mouse serum were significantly up-regulated(P<0.05),the ratio of LC3Ⅱ/Ⅰincreased significantly in cbs^(+/-)mouse liver(P<0.01),p62 expression decreased(P<0.01),and PSMD10 mRNA and protein expression levels increased significantly(P<0.05).For cells,compared with control group,the expression levels of AST and ALT in Hcy group increased significantly(P<0.01),the ratio of LC3Ⅱ/Ⅰincreased(P<0.01),the expression of p62 decreased(P<0.01),and the PSMD10 mRNA and protein expression levels increased significantly(P<0.05).Used PSMD10 siRNA to knock down the expression of PSMD10,compared with Hcy+siNC group,the ratio of LC3Ⅱ/Ⅰdecreased(P<0.01),the expression of p62 increased(P<0.01),and the expression levels of AST and ALT decrea

关 键 词：PSMD10 同型半胱氨酸 肝细胞 自噬 肝损伤 AST ALT 

分 类 号：R-332[医药卫生] R322.47