健肝颗粒诱导线粒体自噬抑制乙肝肝硬化CD4^(+)T细胞凋亡  

Inhibition of apoptosis of CD4^(+)T cells in hepatitis B cirrhosis through inducing mitophagy by Jiangan granules

作　　者：李媛 龙富立[1] 张衎[1] 官志杰[1] 姚凡[1] 彭云鹤 舒发明 唐春回 毛德文[1] 黄古叶[1] LI Yuan;LONG Fuli;ZHANG Kan;GUAN Zhijie;YAO Fan;PENG Yunhe;SHU Faming;TANG Chunhui;MAO Dewen;HUANG Guye(Department of Hepatic Diseases,the First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China;Guangxi Key Laboratory of Translational Medicine for Treating High-Incidence Infectious Diseases with Integrative Medicine,Nanning 530023,China;不详)

机构地区：[1]广西中医药大学第一附属医院肝病科,南宁530023 [2]广西高发传染病中西医结合转化医学重点实验室培育基地,南宁530023 [3]广西巴马瑶族自治县民族医院,广西河池547500

出　　处：《实用医学杂志》2021年第15期1921-1927,共7页The Journal of Practical Medicine

基　　金：广西自然科学基金项目(编号:2018GXNSFAA138077,2019GXNSFBA245018);广西壮族自治区中青年教师基础能力提升项目(编号:2017KY0309)。

摘　　要：目的基于磷酸酶和张力蛋白同源物诱导激酶1(PINK1)/Parkin/活性氧(ROS)通路探讨健肝颗粒抑制乙肝肝硬化(HBC)CD4^(+)T细胞凋亡的作用及机制。方法HBC代偿期患者分为对照组和观察组,健康体检者为正常组,每组50例。对照组给予恩替卡韦治疗,观察组予以健肝颗粒联合恩替卡韦治疗。磁珠分选法筛选各组外周血CD4^(+)T细胞。正常组CD4^(+)T细胞分为Control组、H2O2组和H_(2)O_(2)+CCCP(线粒体自噬激活剂)组。ELISA检测ROS、超氧化物歧化酶(SOD)、丙二醛(MDA)含量,Western blot检测PINK1、Parkin、微管相关蛋白1轻链3-Ⅱ(LC3-Ⅱ)、Bax和Bcl-2表达,流式细胞术检测线粒体膜电位和细胞凋亡率。结果CCCP可逆转H2O2对CD4^(+)T细胞ROS和细胞凋亡率的促进作用和LC3-Ⅱ表达的抑制作用(P<0.05)。健肝颗粒联合恩替卡治疗可下调血清ROS、MDA和升高SOD,以及上调CD4^(+)T细胞PINK1、Parkin、LC3-Ⅱ、Bcl-2表达,下调Bax表达,升高线粒体膜电位,降低细胞凋亡率(P<0.05),且效应优于单用恩替卡韦。结论上调CD4^(+)T细胞线粒体自噬水平可抑制氧化应激和细胞凋亡;健肝颗粒可通过诱导线粒体自噬抑制HBC代偿期患者CD4^(+)T细胞凋亡。Objective Based on the PINK1/Parkin/ROS pathway,to explore the effect of Jiangan Granules on inhibiting the apoptosis of CD4^(+)T cells in liver cirrhosis and its mechanism.Methods Hepatitis B cirrhosis(HBC)patients were divided into control group and observation group,and healthy people were selected as the normal group,with 50 cases in each group.The control group was treated with entecavir,and the observation group with Jiangan Granules combined with entecavir.CD4^(+)T cells in the peripheral blood of each group were screened by magnetic bead sorting method.CD4^(+)T cells in the normal group were divided into control group,H2O2 group and H_(2)O_(2)+CCCP(Mitophagy inducer)group.The contents of ROS,SOD and MDA were detected by ELISA.The expression of PINK1,Parkin,LC3-Ⅱ,Bax and Bcl-2 was detected by Western blot.The mitochondrial membrane potential and apoptosis rate were detected by flow cytometry.Results CCCP could reverse the promoting effect of H2O2 on ROS and apoptosis rate of CD4^(+)T cell and the inhibiting effect of LC3-Ⅱexpression(P<0.05).Jiangan Granules combined with entecavir could down-regulate ROS,MDA and SOD in serum,while up-regulate the expression of PINK 1,Parkin,LC3-Ⅱand Bcl-2 in CD4^(+)T cells and it could also down-regulate the expression of Bax,increase mitochondrial membrane potential and reduce the apoptosis rate(P<0.05).The effect was better than that of entecavir alone.Conclusion Upregulation of CD4^(+)T cell mitophagy level could inhibit oxidative stress and apoptosis.Jiangan Granules could inhibit the apoptosis of CD4^(+)T cell in compensated patients with HBC by inducing mitophagy.

关 键 词：健肝颗粒 线粒体自噬 CD4^(+)T细胞 乙肝肝硬化 氧化应激 

分 类 号：R575[医药卫生—消化系统]