运动介导HSP70延缓增龄性骨骼肌萎缩的机理研究

发布时间：2018-09-03 17:44

【摘要】：目的:通过对自然衰老C57BL/6小鼠进行长期有氧运动干预,探讨增龄性骨骼肌萎缩发生发展过程中细胞凋亡相关蛋白的变化及运动诱导HSP70对其影响,揭示运动预防增龄性骨骼肌萎缩的作用机理。方法:27周龄雄性C57BL/6小鼠50只,经1周适应性喂养后,随机分为年轻对照组(YS组)、老年对照组(OS组)和老年运动组(OE组)。YS组(28周)小鼠经抓力测试后立即处死取材。OE组小鼠采用30周、每周3~4次、每次50分钟、75%VO2max有氧跑台运动训练(速度12米/分钟,坡度0°)。OS组与OE组小鼠喂养至60周龄后,检测抓力等指标后处死。随即分离各组小鼠腓肠肌,称重后检测小鼠骨骼肌萎缩指数(Sarcopenia index,SI),随后提取腓肠肌总蛋白。采用Western Blot检测小鼠腓肠肌组织HSP70,Cleaved Caspase-9蛋白表达水平;采用Co-IP法检测小鼠腓肠肌HSP70与Apaf-1蛋白结合量;采用TUNEL法检测小鼠腓肠肌细胞凋亡指数。结果:(1)SI变化:OS组小鼠腓肠肌SI较YS组显著减少23.09%,而OE组SI与OS组相比显著增加30.48%;(2)小鼠抓力:OS组小鼠抓力/体重比值比YS组显著减少42.74%,OE组比OS组显著增加21.51%;(3)凋亡指数检测:伴随年龄增加,OS组较YS组显著增加475.28%,而有氧运动之后,OE组小鼠凋亡指数较OS组显著减少46.41%;(4)Western Blot结果:OS组Cleaved Caspase-9表达水平较YS组显著增加45.40%,OE组较OS组显著减少33.40%;(5)腓肠肌HSP70与Apaf-1蛋白的结合量:OS组较YS组HSP70与Apaf-1结合量显著下降67.11%,运动后OE组较OS组显著增加306.31%。结论:(1)60周龄小鼠腓肠肌发生增龄性骨骼肌萎缩,30周有氧运动可显著延缓增龄性骨骼肌萎缩;(2)增龄性骨骼肌萎缩可能与骨骼肌凋亡细胞增多有关,30周有氧运动可显著抑制骨骼肌细胞凋亡;(3)30周有氧运动可增强HSP70与Apaf-1结合量,这可能是运动抑制骨骼肌细胞凋亡,延缓增龄性骨骼肌萎缩的机制之一。
[Abstract]:Objective: to investigate the changes of apoptosis-related proteins in aging skeletal muscle atrophy and the effect of exercise induced HSP70 on the aging C57BL/6 mice by long-term aerobic exercise intervention. To explore the mechanism of exercise in preventing aging skeletal muscle atrophy. Methods 50 male C57BL/6 mice at the age of 27 weeks were fed adaptively for one week. They were randomly divided into young control group (YS group), aged control group (OS group) and aged exercise group (OE group) .YS group (28 weeks). The mice in OS group and OE group were fed to 60 weeks of age and killed after 60 weeks of age. Then the gastrocnemius muscle of each group was isolated and the skeletal muscle atrophy index (Sarcopenia index,SI) was measured after weighing and then the total protein of gastrocnemius muscle was extracted. The expression level of HSP70,Cleaved Caspase-9 protein in gastrocnemius muscle was detected by Western Blot, the binding quantity of HSP70 and Apaf-1 protein in gastrocnemius muscle was detected by Co-IP method, and apoptosis index of mouse gastrocnemius muscle was detected by TUNEL method. Results: (1) the change of SI in gastrocnemius muscle of the group of 10% OS was 23.09 less than that of the group of YS, while the SI of the OE group was significantly higher than that of the group of OS, (2) the ratio of grasping power to body weight of the mice in the group of mice grabbing force: OS was significantly lower than that of the group of YS; (3) the ratio of grasping power to body weight in the group of mice grabbing power: OS was significantly lower than that in the group of YS; After aerobic exercise, the apoptotic index of the OE group was significantly lower than that of the OS group (46.41%). (4) the) Western Blot results showed that the Cleaved Caspase-9 expression level in the) Western Blot group was significantly higher than that in the YS group (45.40%) and that in the OS group was significantly lower than that in the OS group. (5) the binding amount of HSP70 to Apaf-1 protein in gastrocnemius muscle in the group of: OS was significantly lower than that in the group of YS (67.11%), and that in the OE group after exercise was significantly higher than that in the group of OS (306.31%). Conclusion: (1) aged skeletal muscle atrophy in 60 week-old mice can be significantly delayed by aerobic exercise for 30 weeks, and (2) aging skeletal muscle atrophy may be related to the increase of apoptotic cells in skeletal muscle. (3) aerobic exercise for 30 weeks increased the binding of HSP70 to Apaf-1. This may be one of the mechanisms of exercise inhibiting the apoptosis of skeletal muscle cells and delaying the aging skeletal muscle atrophy.
【作者单位】：天津市天津医院康复科;天津医科大学基础医学院;天津体育学院运动与健康科学系;
【基金】：天津市应用基础与前沿技术研究计划项目(14JCYBJC27300) 国家自然科学基金(31571220)
【分类号】：R87

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