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

doi:10.13481/j.1671-587x.20190607

摘要/Abstract

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

张恒, 郭彬, 田浩, 李兰, 吴静, 门秀丽. 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[J]. 吉林大学学报(医学版), 2019, 45(06): 1231-1237.

ZHANG Heng, GUO Bin, TIAN Hao, LI Lan, WU Jing, MEN Xiuli. Effect of lithium chloride on apoptosis of islet β cells in rats induced by dexamethasone and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2019, 45(06): 1231-1237.

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