转录因子EB在有氧运动改善高脂饮食诱导小鼠骨骼肌胰岛素抵抗中的作用  

The role of TFEB activated by aerobic exercise on insulin resistance in skeletal muscle of high fat diet mice

作　　者：王平[1] 李佳欣 陈小龙 余新 靳晓鹏 祝明华[2] Wang Ping;Li Jiaxin;Chen Xiaolong;Yu Xin;Jin Xiaopeng;Zhu Minghua(School of Physical Education,Hangzhou Normal University,Hangzhou 311121,China;The Affiliated Hospital,Hangzhou Normal University,Hangzhou 310015,China)

机构地区：[1]杭州师范大学体育学院,杭州311121 [2]杭州师范大学附属医院,杭州310015

出　　处：《中国运动医学杂志》2024年第3期193-204,共12页Chinese Journal of Sports Medicine

基　　金：浙江省哲学社会科学规划课题(19NDJC054YB);浙江省自然科学基金项目(LY24H170002)。

摘　　要：目的:探讨转录因子EB(TFEB)在有氧运动改善骨骼肌胰岛素抵抗中的作用。方法:18只6周龄雄性SPF级别C57BL/6小鼠随机分为普通膳食组(CON组)、高脂膳食组(HFD组)和高脂膳食运动组(HFDE组),每组6只。喂养12周后HFDE组小鼠进行12周跑台运动(60 min/次、5次/周,速度12 m/min,坡度0°)。然后采用腹腔注射葡萄糖耐量试验(IPGTT)和腹腔注射胰岛素耐量试验(IPITT)分别检测小鼠葡萄糖耐量和胰岛素耐量,生化检测小鼠空腹血糖和胰岛素含量、计算胰岛素抵抗指数(HOMA-IR),评价小鼠胰岛素抵抗水平;蛋白质印迹法(WB)检测骨骼肌胞核和胞质磷酸化TFEB(pTFEB)、TFEB蛋白表达、胞质和胞膜葡萄糖转运体4(GLUT4)蛋白表达、骨骼肌胰岛素信号通路磷酸化胰岛素受体底物1(pIRS1)、磷酸化蛋白激酶B(p AKT)、磷脂酰肌醇-3-激酶(PI3K)和pTBC1D4(又名AS160)的蛋白表达;分别对p TFEB、TFEB和胰岛素信号通路相关蛋白做相关性分析。结果:(1)与CON组相比,HFD组小鼠体重、IPGTT、IPITT各个时间点血糖、曲线下面积(AUC)、血清胰岛素、HOMA-IR、胞质pTFEB、总TFEB(TTFEB)和GLUT4蛋白表达均显著增加(P<0.01),但骨骼肌pIRS1、pAKT、PI3K、pTBC1D4、胞膜GLUT4蛋白表达均显著下降(P<0.01),胞核pTFEB和T-TFEB蛋白表达均无显著性差异(P>0.05)。(2)与HFD组相比,HFDE组小鼠体重、IPGTT各个时间点血糖、AUC、IPITT的0 min、30 min、60 min的血糖及AUC、血清胰岛素、HOMA-IR、骨骼肌胞质p TFEB、T-TFEB、GLUT4蛋白表达均显著下降(P<0.05或P<0.01),但pIRS1、pAKT、PI3K、p TBC1D4、胞膜GLUT4和胞核T-TFEB蛋白表达均显著增加(P<0.01)。(3)TFEB与胰岛素信号通路蛋白相关性分析结果显示,胞质p TFEB与pIRS1、PI3K、pAKT和pTBC1D4蛋白表达呈负相关(r=-0.8642,r=-0.7789,r=-0.8946,r=-0.8040,P<0.01),与胞质GLUT4呈正相关(r=0.8532,P<0.01);胞核TFEB与胞膜GLUT4蛋白表达呈正相关(r=0.7744,P<0.01)。结论:高脂膳食可引起小鼠骨Objective To explore the role of transcription factor EB(TFEB) induced by aerobic exercise in improving insulin resistance(IR) of skeletal muscles in high-fat diet mice.Method Eighteen male SPF C57BL/6 mice aged 6 weeks were randomly divided into a common diet group(CON),a high-fat diet group(HFD),and a high-fat diet exercise group(HFDE),each of 6.Both high-fat groups were on high-fat diet for 12 weeks.Then the HFDE group underwent daily 60-minute treadmill exercise with the slope of 0°,at a speed of 12 m/min,5 times per week for 12 weeks.Finally,the glucose tolerance and insulin sensitivity were detected using the intraperitoneal injection glucose tolerance test(IPGTT) and intraperitoneal injection insulin tolerance test(IPITT).The fasting blood glucose and insulin contents were measured by biochemical method,and the IR level was calculated by using the homeostatic model assessment of insulin resistance(HOMA-IR).Moreover,Western blotting was employed to detect the expression of phosphorylated transcription factor EB(pTFEB) and TFEB in skeletal muscle,glucose transporter 4(GLUT4) in cytoplasm and membrane,phosphorylated insulin receptor substrates1(pIRS1),phosphorylated protein kinase B(pAKT),phosphoinositide 3-kinase(PI3K),and phosphorylated akt substrate of 160 k D(p TBC1D4) in skeletal muscle.The correlation of pTFEB and TFEB to insulin signaling pathway-related proteins was analyzed.Results(1) Compared with the CON group,there was a significant increase in the average body weight,IPGTT,blood glucose at each time point of IPGTT,area under curve(AUC),serum insulin,HOMA-IR,protein expressions of pTFEB,total-TFEB and GLUT4 in cytoplasm of the HFD group(P<0.01),but a significant decrease in the average protein expressions of pIRS1,p AKT,PI3K,pTBC1D4 and GLUT4 in cell membrane of skeletal muscles(P<0.01).However,no significant differences were found between the two groups in the average expression of pTFEB and T-TFEB proteins.(2) Compared to the HFD group,a significant increase was found in the average body weight

关 键 词：转录因子EB 高脂膳食 有氧运动 骨骼肌 胰岛素抵抗 

分 类 号：R58[医药卫生—内分泌]