利拉鲁肽通过调节Sestrin2改善棕榈酸诱导的L6骨骼肌细胞胰岛素抵抗  

Liraglutide ameliorates palmitic acid-induced insulin resistance in L6 skeletal muscle cells by regulating Sestrin2

作　　者：田雪 高宇[1] 邢恩鸿[2] 赵丽华[1] 孔莫维 卢亚男[1] 郭晓宇[1] 孙启天 刘晓燕[1] Tian Xue;Gao Yu;Xing Enhong;Zhao Lihua;Kong Mowei;Lu Yanan;Guo Xiaoyu;Sun Qitian;Liu Xiaoyan(Department of Endocrinology,Affiliated Hospital of Chengde Medical University,Chengde 067000,China;Central Laboratory,Affiliated Hospital of Chengde Medical University,Chengde 067000,China)

机构地区：[1]承德医学院附属医院内分泌科,承德067000 [2]承德医学院附属医院中心实验室,承德067000

出　　处：《中华内分泌代谢杂志》2022年第12期1075-1080,共6页Chinese Journal of Endocrinology and Metabolism

基　　金：2020年度河北省医学科学研究课题计划(20200180);河北省科技厅"技术创新引导专项-科技工作会商"项目(2020-2022)。

摘　　要：目的探讨应激诱导蛋白Sestrin2(Sesn2)在利拉鲁肽改善大鼠L6骨骼肌细胞胰岛素抵抗中的作用。方法采用棕榈酸诱导法建立大鼠L6骨骼肌细胞的胰岛素抵抗,将实验细胞分为对照组(Con组)、棕榈酸0.6 mmol/L处理组(PA组)、棕榈酸0.6 mmol/L+利拉鲁肽10 nmol/L处理组(PA+Lir10组)、棕榈酸0.6 mmol/L+利拉鲁肽100 nmol/L处理组(PA+Lir100组)和棕榈酸0.6 mmol/L+利拉鲁肽1000 nmol/L处理组(PA+Lir1000组)。细胞计数试剂盒8(CCK8)法检测各组细胞活性,Western印迹法检测各组葡萄糖转运蛋白4(GLUT4)、蛋白激酶B(Akt)、磷酸化的蛋白激酶B(p-Akt)和Sesn2蛋白的表达。小干扰RNA(siRNA)转染L6细胞,抑制Sesn2的表达后,用棕榈酸0.6 mmol/L和利拉鲁肽100 nmol/L干预细胞,采用Western印迹法检测细胞中Sesn2、Akt、p-Akt、GLUT4蛋白表达水平。结果与Con组相比,PA组细胞存活率明显下降(P<0.05),p-Akt/Akt比值、Sesn2、GLUT4蛋白表达水平下降(P<0.05)。给予利拉鲁肽干预后,PA+Lir100、PA+Lir1000组细胞活性升高(P<0.05),p-Akt/Akt比值、Sesn2、GLUT4蛋白表达水平升高(P<0.05)。抑制Sesn2的表达后,转染si-Sesn2后给予棕榈酸0.6 mmol/L组(PA+si-Sesn2组)和转染si-Sesn2后给予棕榈酸0.6 mmol/L+利拉鲁肽100 nmol/L组(Lir100+PA+si-Sesn2组)的p-Akt/Akt比值及GLUT4蛋白表达水平均明显低于转染阴性组(si-Con组;P<0.05);即使给予利拉鲁肽干预后,与PA+si-Sesn2组相比,Lir100+PA+si-Sesn2组的p-Akt/Akt比值及GLUT4蛋白表达水平无明显升高(P>0.05)。结论棕榈酸可诱导L6细胞p-Akt/Akt比值及GLUT4蛋白表达水平下降。利拉鲁肽通过上调Sesn2表达,使p-Akt/Akt比值、GLUT4蛋白表达水平升高。这一过程可能参与了利拉鲁肽改善棕榈酸引起的L6细胞的胰岛素抵抗。Objective To investigate the role of stress-inducible protein Sestrin2(Sesn2)in the improvement of insulin resistance in rat L6 skeletal muscle cells treated with liraglutide.Methods The establishment of insulin resistance model of rat L6 skeletal muscle cells was induced by palmitate.The experimental cells were divided into control group(Con group),palmitate 0.6 mmol/L treatment group(PA group),palmitate 0.6 mmol/L+liraglutide 10 nmol/L treatment group(PA+Lir10 group),palmitate 0.6 mmol/L+liraglutide 100 nmol/L treatment group(PA+Lir100 group),and palmitate 0.6 mmol/L+liraglutide 1000 nmol/L treatment group(PA+Lir1000 group).The cell counting kit 8(CCK8)method was used to detect the cell activity in each group.Western blotting was used to detect the expression levels of glucose transporter 4(GLUT4),protein kinase B(Akt),phosphorylated protein kinase B(p-Akt),and Sesn2 protein in L6 cells.L6 cells were transfected with siRNA to inhibit the expression of Sesn2.The cells were treated with palmitate and liraglutide.Western blotting was used to detect the expression levels of Sesn2,Akt,p-Akt,and GLUT4 protein in L6 cells.Results Compared with Con group,the cell survival rate,p-Akt/Akt ratio,Sesn2,and GLUT4 protein expression in PA group decreased significantly(P<0.05).After liraglutide intervention,the cell activity,p-Akt/Akt ratio,Sesn2,and GLUT4 protein expression of PA+Lir100 and PA+Lir1000 groups was increased(P<0.05).After inhibiting the expression of Sesn2,p-Akt/Akt ratio and GLUT4 protein in transfected si-Sesn2 and treated with 0.6 mmol/L palmitate group(PA+si-Sesn2 group)and transfected si-Sesn2 and treated with 0.6 mmol/L palmitate+liraglutide 100 nmol/L group(Lir100+PA+si-Sesn2 group)were significantly lower than those in transfection negative group(si-Con group;P<0.05).Even after liraglutide intervention,compared with PA+si-Sesn2 group,p-Akt/Akt ratio and GLUT4 protein expression level were not significantly increased in Lir100+PA+si-Sesn2 group(P>0.05).Conclusions Palmitate could induce the decrease of p

关 键 词：利拉鲁肽 胰岛素抵抗 L6骨骼肌细胞 棕榈酸 Sestrin2 

分 类 号：R587.1[医药卫生—内分泌]