实用医学杂志 ›› 2021, Vol. 37 ›› Issue (15): 1921-1927.doi: 10.3969/j.issn.1006⁃5725.2021.15.003

• 基础研究 • 上一篇    下一篇

健肝颗粒诱导线粒体自噬抑制乙肝肝硬化CD4+T细胞凋亡

李媛1,2 ,龙富立1, 张衎1, 官志杰1, 姚凡1, 彭云鹤1, 舒发明1, 唐春回3, 毛德文1, 黄古叶1   

  1. 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)

  • 出版日期:2021-08-10 发布日期:2021-08-10
  • 通讯作者: 黄古叶 E⁃mail:huangguye@163.com
  • 基金资助:
    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)

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

LI Yuan*,LONG Fuli,ZHANG Kan,GUAN Zhijie,YAO Fan,PENG Yunhe,SHU Faming,TANG Chunhui,MAO Dewen,HUANG Guye.   

  1.  Department of Hepatic Diseases,the First Affiliated Hospital of Guangxi Uni⁃ versity of Chinese Medicine,Nanning 530023,China;* Guangxi Key Laboratory of Translational Medicine for Treat⁃ ing High⁃Incidence Infectious Diseases with Integrative Medicine,Nanning 530023,China

  • Online:2021-08-10 Published:2021-08-10
  • Contact: HUANG Guye E⁃mail:huangguye@163.com
  • Supported by:


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H2O2+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 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 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。

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

Abstract:

Objective Based on the PINK1/Parkin/ROS pathway,to explore the effect of Jiangan Gran⁃ ules on inhibiting the apoptosis of CD4+ T cells in liver cirrhosis and its mechanism. Methods Hepatitis B cirrho⁃ sis(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 H2O2+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 mem⁃ brane potential and apoptosis rate were detected by flow cytometry. Results CCCP could reverse the promoting ef⁃ fect of H2O2 on ROS and apoptosis rate of CD4+ T cell and the inhibiting effect of LC3⁃Ⅱ expression(P<0.05). Ji⁃ angan Granules combined with entecavir could down ⁃ regulate ROS,MDA and SOD in serum,while up ⁃ regulate the expression of PINK1,Parkin,LC3⁃Ⅱ and 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 HBCby inducing mitophagy.

Key words:

Jiangan granules, mitophagy, CD4+ T cells, hepatitis B cirrhosis, oxidative stress