慢病毒介导沉默SOCS3基因对糖尿病大鼠相关胰岛素通路蛋白表达的影响

孙志兵; 杨娟

doi:10.13898/j.cnki.issn.1000-2200.2020.05.004

慢病毒介导沉默SOCS3基因对糖尿病大鼠相关胰岛素通路蛋白表达的影响

孙志兵,
杨娟

Effect of lentivirus-mediated silencing SOCS3 gene on the expression of related insulin pathway proteins in diabetic rats

摘要

摘要:
目的构建糖尿病大鼠模型，探讨慢病毒介导沉默细胞因子信号转导抑制因子3(suppressors of cytokine signaling 3，SOCS3)基因对大鼠相关胰岛素通路蛋白表达的影响。
方法选取SD大鼠40只，构建2型糖尿病大鼠模型成功后，随机分为空白对照组、空白载体组和观察组，各10只。空白对照组不进行其他处理，空白载体组通过大鼠尾部静脉注射空慢病毒载体，观察组通过大鼠尾部静脉注射SOCS3 RNAi慢病毒载体。注射载体4周后，测定3组大鼠血糖、血脂、胰岛素水平；采用实时荧光定量聚合酶链式反应法检测SOCS3、IRS1、IRS2、STAT5b、JAK2和STAT3 mRNA表达，Western blotting法检测SOCS3、IRS1、pIRS1、IRS2、pIRS2、STAT5b、pSTAT5b、JAK2、pJAK2、STAT3、pSTAT3蛋白表达。
结果注射载体4周后，观察组大鼠空腹血糖、胰岛素、三酰甘油、总胆固醇、低密度脂蛋白胆固醇水平均明显低于空白对照组和空白载体组(P＜0.01)，高密度脂蛋白胆固醇水平均明显高于空白对照组和空白载体组(P＜0.01)；观察组SOCS3 mRNA和STAT5b mRNA表达均低于空白对照组和空白载体组(P＜0.01)，IRS1 mRNA、IRS2 mRNA和JAK2 mRNA表达均高于空白对照组和空白载体组(P＜0.05～P＜0.01)，3组STAT3 mRNA差异无统计学意义(P>0.05)；观察组SOCS3、STAT5b、pSTAT5b表达均明显低于空白对照组和空白载体组(P＜0.01)，IRS1、pIRS1、IRS2、pIRS2、JAK2、pJAK2、pSTAT3蛋白表达均明显高于空白对照组和空白载体组(P＜0.01)，3组STAT3蛋白表达差异无统计学意义(P＞0.05)。
结论沉默SOCS3基因可降低糖尿病大鼠血糖水平，SOCS3基因可能通过相关信号通路介导胰岛素抵抗。

Abstract:
ObjectiveTo construct the diabetic rat model, and investigate the effects of lentivirus-mediated silencing the suppressors of cytokine signaling 3(SOCS3) gene on the expression of related insulin pathway proteins in diabetic rats.
MethodsForty SD rats model of type 2 diabetes were constructed, and randomly divided into the blank control group, blank vector group and observation group(10 rats each group).The blank control group was not treated, the empty lentiviral vector was injected into the tail of rats of blank vector group, and the SOCS3 RNAi lentiviral vector was injected into the tail of rats of observation group.The levels of blood glucose, blood lipid and insulin in three groups were measured after 4 weeks of injection.The expression levels of SOCS3, IRS1, IRS2, STAT5b, JAK2 and STAT3 mRNA were detected using RT-PCR, and the expression levels of SOCS3, IRS1, pIRS1, IRS2, pIRS2, STAT5b, pSTAT5b, JAK2, pJAK2, STAT3 and pSTAT3 protein were detected using Western blotting.
ResultsAfter 4 weeks of injection, the levels of fasting blood glucose, insulin, TG, LDL-C and TC in observation group were significantly lower than those in blank control group and blank vector group(P＜0.01), and the level of HDL-C in observation group was significantly higher than that in blank control group and blank vector group(P＜0.01).The expression levels of SOCS3 and STAT5b gene mRNA in observation group were lower than those in blank control group and blank vector group(P＜0.01), the expression levels of IRS1, IRS2 and JAK2 mRNA in observation group were higher than those in blank control group and blank vector group(P＜0.05 to P＜0.01), and the differences of the expression level of STAT3 mRNA among three groups were not statistically significant(P>0.05).The expression levels of SOCS3, STAT5b and pSTAT5b protein in observation group were significantly lower than those in blank control group and blank vector group(P＜0.01), the expression levels of IRS1, pIRS1, IRS2, pIRS2, JAK2, pJAK2 and pSTAT3 protein in observation group were significantly higher than those in blank control group and blank vector group(P＜0.01), and the differences of the levels of STAT3 protein among three groups were not statistically significant(P＞0.05).
ConclusionsSilencing the SOCS3 gene can reduce the blood glucose level in diabetic rats, and the SOCS3 gene may mediate insulin resistance through related pathways.

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