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李慧阁,牛燕媚,刘彦辉,苏照鹏,傅 力.小鼠骨骼肌细胞蛋白激酶B底物160在有氧运动促进葡萄糖转运体4转运中的作用[J].中国康复医学杂志,2010,25(6):501~506
小鼠骨骼肌细胞蛋白激酶B底物160在有氧运动促进葡萄糖转运体4转运中的作用    点此下载全文
李慧阁  牛燕媚  刘彦辉  苏照鹏  傅 力
天津体育学院健康与运动科学系,300381
基金项目:国家自然科学基金(30871213);天津市应用基础和前沿技术重点项目(09JCZDJC17400);天津市重大攻关项目(05YFGDSF02100)
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摘要:
      摘要 目的:通过研究有氧运动对C57BL/6 小鼠骨骼肌细胞蛋白激酶B (PKB/Akt) 底物蛋白 160 (AS160) 和葡萄糖转运体 4 (GLUT4) 表达的影响,探讨 AS160 在调节细胞葡萄糖转运过程中的作用及有氧运动影响骨骼肌细胞葡萄糖代谢的生物学机制。 方法:20只16周龄、雄性 C57BL/6 小鼠,随机分为安静组 (NC,n=10) 和运动组 (NE,n=10);运动组进行为期 6周、75% VO2max 强度的有氧跑台训练。6周训练结束后24h,动物麻醉后取材。采用逆转录聚合酶链反应(RT-PCR) 检测骨骼肌组织 AS160、GLUT4 基因表达;免疫印迹杂交 (Western blot) 和免疫荧光染色 (IF) 分别检测骨骼肌细胞 AS160、pAS160-Thr642和 GLUT4 蛋白表达及定位。 结果:与NC组相比,NE 组小鼠骨骼肌细胞 AS160 mRNA 和蛋白表达差异无显著性,但 pAS160-Thr642表达显著增加;NE 组小鼠骨骼肌细胞 GLUT4 mRNA 和蛋白表达较 NC 组显著增加,GLUT4 向细胞膜迁移明显增加。 结论:有氧运动显著增强骨骼肌细胞 pAS160-Thr642的活性,增强 GLUT4 表达及促进 GLUT4 向细胞膜转移,促进骨骼肌细胞对葡萄糖的摄取和利用。
关键词:蛋白激酶B底物160  有氧运动  葡萄糖转运体4
The potential role of protein kinaseB/Akt substrate 160 in the process of aerobic exercises improving glucose transporter 4 trafficking    Download Fulltext
Tianjin University of Sport,300381
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Abstract:
      Abstract Objective: To study the regulatory effect of AKt substrate160(AS160) in aerobic exercises improving skeletal muscle glucose transport, and to explore the expression of AS160 and glucose transporter4(GLUT4) gene and protein in C57BL/6 mice skeletal muscle. Method: Twenty male aged 16-week C57BL/6 mice were divided into control group (NC) and exercises group (NE) randomly. NE mice underwent 6-week treadmill training. Exercises training consisted of motorized treadmill running for 1h/d, 5 d/week at an intensity of 75%VO2max; reverse transcription-polymerase chain reaction(RT-PCR) and Western blot were applied to detect AS160 and GLUT4 gene and AS160, pAS160-Thr642 and GLUT4 protein expressions in skeletal muscle. Immunofluorescence was used to detect the expressions and locations of AS160,pAS160-Thr642 and GLUT4. Result: The expressions of AS160 gene and protein had no difference between two groups, however there was significant increment in pAS160-Thr642 expression in NE group compared to that in NC group; while the expressions of both GLUT4 mRNA and protein increased significantly in NE group compared to that in NC group. Moreover, immunofluorescence staining further supported the Western blot results, and there is significant increase of GLUT4 translocating towards cell membrane. Conclusion: Aerobic exercises can promote the activity of Akt/AS160 signaling pathway in mouse skeletal muscle by increasing the expression of pAS160-Thr642, and then elevated the expressions of GLUT4 gene and protein, promote GLU4 translocating towards cell membrane and skeleton muscle cells intaking and utilizing glucose.
Keywords:protein kinase B/AKt substrate160  aerobic exercises  glucose transporter 4
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