摘要
目的探究乙型肝炎病毒(HBV)全x蛋白(HBwx)诱导肝星状细胞(HSCs)炎症反应和促进肝细胞凋亡的作用及其机制。方法取LX2细胞,分为对照组、脂多糖(LPS)处理组、转染HBwx处理组和LPS联合转染HBwx处理组。采用Western blot法检测自噬相关蛋白p62和LC3及炎症相关蛋白NLRP3和pro-IL表达,采用ELISA法检测培养上清IL-1β水平;收集上述各组LX2细胞培养上清刺激HL7702细胞培养,使用流式细胞仪检测HL7702细胞凋亡。结果经HBwx质粒转染的LX2细胞在细胞质表达HBwx蛋白;与正常组比,LPS组、HBwx组和LPS联合HBwx组LC3表达显著减低,而p62表达显著增强(P<0.05);与正常组比,LPS组、HBwx组和LPS联合HBwx组NLRP3和pro-IL1表达显著增强(P<0.05);与正常组比,LPS组、HBwx组和LPS联合HBwx组IL-1β分泌显著增多(P<0.05);与正常组比,LPS、HBwx和LPS联合HBwx处理HL-7702细胞凋亡水平显著增强(P<0.05);与LPS组或HBwx组比,LPS联合HBwx处理HL-7702细胞凋亡水平进一步增强(P<0.05)。结论HBwx作用于HSCs抑制自噬并促进细胞炎症反应,HSCs分泌IL-1β,诱导肝细胞凋亡。
Objective The purpose of this experiment was to investigate hepatitis B whole x(HBwx)gene promotes inflammation reaction of hepatic stellate cells(HSCs)and triggers hepatocyte apoptosis in vitro.Methods The LX2 cells were divided into control,lipopolysaccharide(LPS)-,HBwx transfected and LPS and HBwx transfection combination-intervened groups.The cell p62 and LC3 as well as NOD-like receptor protein 3(NLRP3)and pro-IL expression were detected by Western blot,and supernatant IL-1β levels were detected by ELISA.The HL7702 hepatocytes were co-cultured by the supernatants of LX2 cells intervened mentioned above.The cell apoptosis was detected by FCM.Results The HBwx protein was clearly expressed in the cytoplasm of transfected LX2 cells;the LC3 expression decreased,while p62 expression increased in LPS-,HBwx-and LPS and HBwx combination-intervened groups as compared to in the control(P<0.05);the NLRP3 and pro-IL expression increased in LPS-,HBwx-and LPS and HBwx combination-intervened groups as compared to in the control(P<0.05);the supernatant IL-1β levels increased in LPS-,HBwx-and LPS and HBwx combination-intervened groups as compared to in the control(P<0.05);the apoptosis of HL-7702 cells significantly increased in LPS-,HBwx-and especially in LPS and HBwx combination-intervened groups as compared to in the control(P<0.05).Conclusions HBwx promotes inflammatory reaction in HSCs by inhibition of cell autophagy and induces hepatocyte apoptosis.
作者
何亚娟
王飞
姚耐娟
吴宇超
田臻
He Yajuan;Wang Fei;Yao Naijuan(Department of Ultrasound,First Affiliated Hospital,Xi'an Jiaotong University,Xi'an 710061,Shaanxi Province,China)
出处
《实用肝脏病杂志》
CAS
2022年第3期323-326,共4页
Journal of Practical Hepatology
基金
国家自然科学基金资助项目(编号:81800548)。
