胰岛素对谷胱甘肽过氧化物酶表达的影响及其意义

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摘要目的探讨胰岛素对谷胱甘肽过氧化物酶（GSH-Px）表达的影响及其意义。方法用鸡尾酒法诱导3T3-L1前脂肪细胞分化，油红O染色鉴定其分化程度；3T3-L1脂肪细胞用500 nmol/L胰岛素处理1 h后用实时荧光定量PCR方法检测GSH-Px1、GSH-Px4 和GSH-Px7 mRNA表达水平；3T3-L1脂肪细胞用GSH-Px抑制剂Mercaptosuccinate（MS，10、20 mmol/L）预处理30 min后加或不加500 nmol/L胰岛素刺激4 h，用Western blot法检测葡萄糖转运蛋白4（GLUT4）蛋白表达水平。结果 3T3-L1前脂肪细胞呈长梭形；诱导分化后第5天可见细胞逐渐变为球型，胞内可见细小脂滴；诱导分化后第8天可见细胞进一步变为球形且胞内脂滴明显增多。胰岛素组GSH-Px1、GSH-Px4、GSH-Px7 mRNA相对表达量低于对照组，差异有统计学意义（P ＜ 0.05）；存在或不存在胰岛素条件下，10 mmol/L MS处理组与MS未处理组GLUT4蛋白表达比较，差异无统计学意义（P ＞ 0.05）；20 mmol/L MS处理组GLUT4蛋白表达低于MS未处理组，差异有统计学意义（P ＜ 0.05）。结论长时间胰岛素刺激抑制GSH-Px mRNA的表达，用MS抑制GSH-Px活性负性调控GLUT4蛋白表达。

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	马金辉1 刘洁1 李媛媛2 宋彦坤3 郑立双4 檀迎会4 郗昕4

关键词 ：胰岛素, 葡萄糖转运蛋白4, 3T3-L1脂肪细胞, 谷胱甘肽过氧化物酶

Abstract：Objective To explore the effects of insulin on glutathione peroxidase (GSH-Px) expression and its significance. Methods The differentiation of 3T3-L1 preadipocytes were induced by cocktail method and the degree of differentiation was identified by oil red O staining; after 3T3-L1 adipocytes were treated with 500 nmol/L insulin for 1 h, the mRNA expression levels of GSH-PX1, GSH-PX4, and GSH-PX7 were detected by real-time quantitative PCR; 3T3-L1 adipocytes were pretreated with GSH-Px inhibitor Mercaptosuccinate (MS, 10, 20 mmol/L) for 30 min and then stimulated with or without 500 nmol/L insulin for 4 h, the expression level of glucose transporter 4 (GLUT4) protein was detected by Western blot. Results 3T3-L1 preadipocytes were spindle-shaped; and the cells gradually became spheroid and small lipid droplets were observed in the cells on the 5th day after induced differentiation; the cells further became spherical and the number of intracellular lipid droplets increased significantly on the 8th day after induced differentiation. The mRNA relative expression levels of GSH-Px1，GSH-Px4, and GSH-Px7 in insulin group were lower than those in control group, and the differences were statistically significant (P < 0.05); with or without insulin, there was no significant in expression of GLUT4 protein between the 10 mmol/L MS treatment group and the MS untreated group (P ＞ 0.05); the level of GLUT4 protein in 20 mmol/L MS treated group was lower than that in MS untreated group, and the difference was statistically significant (P < 0.05). Conclusion Long-term insulin stimulation inhibited GSH-Px mRNA expression, and MS inhibition of GSH-Px activity negatively regulated GLUT4 protein expression.

Key words： Insulin Glucose transporter 4 3T3-L1 adipocytes Glutathione peroxidase

基金资助:国家自然科学基金资助项目（31701238）；
河北省重点研发计划健康医疗与生物医药专项项目（182777105D）；
河北省医学科学研究重点课题计划项目（20180706）；河北省保定市科技计划项目（2141ZF317）。

通讯作者: 郗昕（1983-），女，博士，主要从事糖尿病基础医学研究。

作者简介: 马金辉（1982-），男，博士，主要从事糖尿病研究。

引用本文:

马金辉1 刘洁1 李媛媛2 宋彦坤3 郑立双4 檀迎会4 郗昕4. 胰岛素对谷胱甘肽过氧化物酶表达的影响及其意义[J]. 中国医药导报, 2022, 19(15): 4-7.
MA Jinhui1 LIU Jie1 LI Yuanyuan2 SONG Yankun3 ZHENG Lishuang4 TAN Yinghui4 XI Xin4. Effects of insulin on glutathione peroxidase expression and its significance. 中国医药导报, 2022, 19(15): 4-7.

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