吉林大学学报(医学版) ›› 2019, Vol. 45 ›› Issue (06): 1231-1237.doi: 10.13481/j.1671-587x.20190607

• 基础研究 • 上一篇    下一篇

氯化锂对地塞米松诱导的大鼠胰岛β细胞凋亡的影响及其机制

张恒, 郭彬, 田浩, 李兰, 吴静, 门秀丽   

  1. 华北理工大学基础医学院河北省慢性疾病重点实验室, 河北 唐山 063210
  • 收稿日期:2019-01-10 出版日期:2019-12-05 发布日期:2019-12-05
  • 通讯作者: 门秀丽,教授,硕士研究生导师(Tel:0315-8805518,E-mail:xiulimen@126.com) E-mail:xiulimen@126.com
  • 作者简介:张恒(1994-),男,湖北省赤壁市人,在读医学硕士,主要从事慢性病病理生理学方面的研究。
  • 基金资助:
    国家自然科学基金资助课题(81370918)

Effect of lithium chloride on apoptosis of islet β cells in rats induced by dexamethasone and its mechanism

ZHANG Heng, GUO Bin, TIAN Hao, LI Lan, WU Jing, MEN Xiuli   

  1. Key Laboratory of Chronic Diseases, Hebei Province, School of Basic Medical Sciences, North China University of Science and Technology, Tangshan 063000, China
  • Received:2019-01-10 Online:2019-12-05 Published:2019-12-05

摘要: 目的:建立地塞米松(Dex)诱导的大鼠胰岛INS-1细胞凋亡模型,探讨氯化锂(LiCl)对Dex诱导的胰岛β细胞凋亡的影响及其可能机制。方法:将INS-1细胞分为对照组、0.1 μmol·L-1Dex组和LiCl+0.1 μmol·L-1Dex组。TUNEL染色和Annexin Ⅴ/PI染色法检测各组INS-1细胞凋亡率,Real-time PCR法检测各组INS-1细胞中超氧化物歧化酶(SOD)、诱导型-氧化氮合酶(iNOS)、NADPH氧化酶4(Nox4)、NADPH oxidase (p47phox)和糖原合成酶激酶3β(GSK-3β) mRNA表达水平,Western blotting法检测各组INS-1细胞中GSK-3β、p-GSK-3、SOD、iNOS和Nox4蛋白表达水平,GENMED试剂盒检测各组NIS-1细胞中活性氧(ROS)水平,Griess法检测各组INS-1细胞中一氧化氮(NO)水平。结果:与对照组比较,0.1 μmol·L-1Dex组INS-1细胞凋亡率升高(P<0.05),SOD mRNA和p-GSK-3β蛋白表达水平降低(P<0.05),细胞中Nox4、p47phox、iNOS mRNA表达水平升高(P<0.05),细胞中ROS和NO水平升高(P<0.05);与0.1 μmol·L-1Dex组比较,LiCl+0.1μmol·L-1Dex组INS-1细胞凋亡率降低(P<0.05),SOD mRNA和p-GSK-3β蛋白表达水平升高(P<0.05),细胞中Nox4、p47phox和iNOS mRNA表达水平降低(P<0.05或P<0.01),细胞中ROS和NO水平降低(P<0.05)。结论:Dex可诱导大鼠胰岛β细胞凋亡,LiCl可通过抑制GSK-3β活性减轻Dex诱导的胰岛β细胞凋亡。

关键词: 氯化锂, 糖原合成酶激酶3β, 地塞米松, 细胞凋亡, INS-1细胞

Abstract: Objective: To construct the apoptosis model of islet INS-1 cells of the rats induced by dexamethasone(Dex),and to investigate the effect of lithium chloride(LiCl) on the apoptosis of the islet β cells induced by Dex and its possible mechanism. Methods: The INS-1 cells were divided into control group,0.1 μmol·L-1 Dex group and LiCl+0.1 μmol·L-1 Dex group. TUNEL staining and Annexin Ⅴ/PI staining methods were used to detect the apoptotic rates of the INS-1 cells in various groups;Real-time PCR method was used to detect the expression levels of superoxide dismutase (SOD),inducible-nitric oxidesynthase (iNOS),NADPH oxidase 4 (Nox4),NADPH oxidase (p47phox),and glycogen-synthase kinase-3β (GSK3β) mRNA in the INS-1 cells in various groups;Western blotting method was used to detect the expression levels of GSK-3β,p-GSK-3β,SOD,iNOS and Nox4 proteins in the INS-1 cells in various groups;GENMED Kit was used to detect the levels of reactive oxygen species (ROS) in the INS-1 cells in various groups,and Griess method was used to detect the levels of nitric oxide (NO) in the INS-1 cells in various groups. Results: Compared with control group,the apoptotic rate of INS-1 cells in 0.1 μmol·L-1 Dex group was significantly increased(P<0.05),the expression levels of SOD mRNA and p-GSK-3β proteins were decreased(P<0.05),the expression levels of Nox4,p47phox,iNOS mRNA were increased(P<0.05),and the expression levels of ROS and NO in the INS-1 cells were significantly increased (P<0.05); compared with 0.1 μmol·L-1 Dex group,the apoptotic rate of INS-1 cells in LiCl+0.1 μmol·L-1 Dex group was significantly decreased(P<0.05),the expression levels of SOD mRNA and p-GSK-3β protein were increased(P<0.05),the expression levels of Nox4,p47phox and iNOS mRNA were decreased(P<0.05 or P<0.01),and the levels of ROS and NO were decreased (P<0.05). Conclusion: Dex can induce the apoptosis of the islet β cells of the rats.LiCl can attenuate the apoptosis of dexamethasone-induced islet β cells by inhibiting the activity of GSK-3β.

Key words: lithium chloride, glycogen synthase kinase-3β, dexamethasone, apoptosis, INS-1 cells

中图分类号: 

  • R329.28