基础医学与临床 ›› 2019, Vol. 39 ›› Issue (5): 617-622.

• 研究论文 •    下一篇

乙肝病毒X蛋白抑制小鼠足细胞系MPC5增殖并促进其凋亡

雷晓燕,孙永红,陈星星,崔维静,刘璟   

  1. 甘肃省人民医院
  • 收稿日期:2018-04-09 修回日期:2018-07-25 出版日期:2019-05-05 发布日期:2019-04-24
  • 通讯作者: 雷晓燕 E-mail:leixiaoyan601@163.com
  • 基金资助:
    国家自然科学基金项目;甘肃省自然科学基金资助项目

Hepatitis B virus X protein inhibits the proliferation and promotes the apoptosis of mouse foot cell line MPC5

  • Received:2018-04-09 Revised:2018-07-25 Online:2019-05-05 Published:2019-04-24

摘要: 目的 探讨乙肝病毒X蛋白(HBx)对条件永生型小鼠足细胞系(MPC5) 增殖与凋亡的影响。方法 采用携带HBx基因的pEX质粒转染MPC5细胞,实时荧光定量PCR(qRT-PCR)验证转染效率。实验分3组,即HBx转染组(MPC5-pEX-HBx group):脂质体+HBx-质粒; 阴性对照组(MPC5-pEX-neo group):脂质体+空质粒;空白对照组(MPC5 group):脂质体组。MTT法检测HBx对足细胞存活率的影响;流式细胞术检测各组细胞凋亡情况;Western blot法检测各组细胞中Nerhrin、STAT3、JAK2蛋白表达水平。结果 pEX-HBx 转染MPC5细胞后48小时HBx表达最高。HBx转染组中Nephrin蛋白表达水平显著低于空白对照及阴性对照(P值均<0.01)。转染HBx基因后足细胞增殖明显受抑,同时凋亡率显著增高(P值均<0.01),此外HBx转染组STAT3及JAK2蛋白表达较空白对照及阴性对照组均显著增高(P值均<0.01)。结论 HBx可下调足细胞中nephrin蛋白表达,抑制足细胞增殖,并促进其凋亡的发生,作用机制可能与STAT3/JAK2信号通路活化有关

关键词: 乙肝病毒X蛋白, 足细胞, 增殖, 凋亡

Abstract: Objective To investigate the effect of hepatitis b virus X (HBx) on the proliferation and apoptosis of condition immortalized mouse foot cell line(MPC5).Methods We used pEX plasmid with HBx gene transfected MPC5 cells,and quantitative Real-time PCR(qRT-PCR) tested the transfection efficiency. The experiment was divided into three groups: HBx transfection group (MPC5-pEX-HBx group):liposome+HBx-plasmid; Negative control group (MPC5-pEX-neo group) : liposome+empty plasmid; Blank control group (MPC5 group) : liposome. Survival rate of each group was determined by MTT method. The cell apotosis was analyzed by flow cytometry. Western blot was employed to detect the Nerhrin、STAT3、JAK2 proteins level. Results There was the highest expression of HBx in 48 hours after transfection with MPC5 cells. The expression level of Nephrin protein in HBx transfection group was lower than that in blank and negative control( P<0.01). After transfection of HBx gene, the podocyte proliferation was inhibited, and the apoptosis rate was increased significantly (P<0.01), what’s more, STAT3 and JAK2 expression were all increased than control groups(P<0.01). Conclusion HBx can reduce the nephrin expression in podocyte, and inhibit it proliferation and promote which apoptosis, this mechanism may be related to the activation of STAT3/JAK2 signaling pathway.

Key words: hepatitis b virus X, podocyte, proliferation, apoptosis