游泳运动对糖尿病小鼠骨骼肌AMPK/SIRT1/NF-κB炎症信号通路的影响

张坦; 崔迪; 张喆; 孙易; 丁树哲

doi:10.16469/j.css.201609006

游泳运动对糖尿病小鼠骨骼肌AMPK/SIRT1/NF-κB炎症信号通路的影响

The Effect of Swimming Exercise on Inflammation via AMPK/SIRT1/NF-κB Signaling Pathway in the Skeletal Muscles of Diabetic Mice

摘要

摘要: 目的:探讨耐力运动对糖尿病小鼠骨骼肌AMPK/SIRT1/NF-κB炎症信号通路及相关炎症因子的影响,以其从骨骼肌慢性炎症的角度为肥胖、糖尿病等代谢类疾病的防治提供新的研究靶向。方法:采用4周高脂膳食喂养加注射链脲佐菌素(Streptozotocin,STZ)的方法构建糖尿病小鼠模型,建模成功后随机分为安静对照组(C)、运动对照组(E)、糖尿病安静组(D)、糖尿病运动组(DE)。E组和DE组进行6周游泳耐力运动,1h/天,5天/周。末次运动结束,空腹12h后处死小鼠并采样。RT-PCR及Western Blotting技术检测相关基因的mRNA及蛋白表达水平。结果:1)与C组相比,D组小鼠体重极显著降低(P<0.01),空腹血糖显著升高(P<0.01);与C组相比,E组小鼠体重显著降低(P<0.05);与D组相比,DE组小鼠空腹血糖显著降低(P<0.05);与E组相比,DE组小鼠体重极显著性降低(P<0.01),空腹血糖极显著性升高(P<0.01)。2)与C组相比,D组小鼠IL-10mRNA表达极显著性降低(P<0.01),E组IL-6 mRNA表达显著性升高(P<0.05),IL-10 mRNA表达极显著性升高(P<0.01);与D组相比,DE组TNF-αmRNA表达显著降低(P<0.05),IL-10 mRNA表达极显著性升高(P<0.01);与E组相比,DE组IL-6 mRNA表达显著性下降(P<0.05)。3)与C组相比,D组AMPK蛋白表达和AMPK活性显著性下降(P<0.05),NF-κBp65 mRNA表达显著性升高(P<0.05);与D组相比,DE组AMPKα2及SIRT1 mRNA表达极显著性升高(P<0.01),AMPK及p-AMPK蛋白表达和AMPK活性显著性升高(P<0.05),NF-κBp65蛋白表达极显著性下降(P<0.01);与E组相比,DE组SIRT1 mRNA表达极显著性升高(P<0.01)。结论:长期耐力运动可抑制糖尿病小鼠骨骼肌中促炎因子的基因表达,促进抗炎因子的基因表达,同时,AMPK/SIRT1/NF-κB炎症信号通路的被抑制效应得到缓解。

Abstract: Objective:The purpose of the present study was to investigate the effect of endurance exercise on inflammatory factors and corresponding signaling pathways AMPK/SIRT1/NF-κB in the skeletal muscles of diabetic mice,hoping to provide new evidence for the prevention and treatment of obesity and diabetes from the perspective of inflammation of the skeletal muscle.Methods:The diabetic mice were established by feeding with high-fat diet for 4 weeks,and then injected with Streptozotocin.All the mice were randomly chosen and assigned to either control group(C,n=8)or exercise group(E,n=8),diabetic group(D,n=8)or diabetic exercise group(DE,n=8),Groups E and DE were then interfered with 6-week swimming exercise of moderate intensity,one hour per day,five days per week.RT-PCR was used to measure the mRNA expression of TNF-α,CCL2,IL-6,IL-10,AMPKα1,AMPKα2,SIRT1 and NF-κBp65.Western Blotting was used to measure the protein level of AMPK,p-AMPK,SIRT1,NF-кBp65 and AC-NF-кBp65.Results:1)Compared with group C,the weight of mice in group D decreased significantly(P<0.01),meanwhile the fasting blood glucose increased significantly(P<0.01);Compared with group C,the weight of mice in group E decreased significantly(P<0.05);Compared with group D,the fasting blood glucose in group DE decreased significantly(P<0.05);Compared with group E,the weight of mice in group DE decreased significantly,while the fasting blood glucose increased significantly(P<0.01).2)Compared with group C,the IL-10 mRNA expression in group D was decreased significantly(P<0.01);while,the IL-6 mRNA expression in group E was increased significantly(P<0.05);Compared with group D,the TNF-αmRNA in group DE was decreased significantly(P<0.05),the IL-10 mRNA in group DE was increased significantly(P<0.01);Compared with group E,the IL-6 mRNA of group DE was decreased significantly(P<0.05).3)Compared with group C,the protein expression and activity of AMPK were decreased significantly(P<0.05)in group D,while the mRNA of NF-κBp65 was increased significantly(P<0.01)in group D;Compared with group D,the mRNA of AMPKα2 and SIRT1 were increased significantly(P<0.01)in group DE,the protein expression of AMPK、p-AMPK and the activity of AMPK were increased significantly(P<0.05),while the protein expression of NF-κBp65 was decreased significantly(P<0.01);Compared with group E,the mRNA of SIRT1 increased significantly(P<0.01).Conclusions:6 weeks regular swimming exercise can promote the mRNA expression of anti-inflammatory factors,while the pro-inflammatory factors were repressed in skeletal muscles of diabetic mice.What's more,exercise can reverse relieve the suppression of AMPK/SIRT1/NF-кB by diabetes.

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