【摘要】：目的:通過對自然衰老C57BL/6小鼠進(jìn)行長期有氧運(yùn)動干預(yù),探討增齡性骨骼肌萎縮發(fā)生發(fā)展過程中細(xì)胞凋亡相關(guān)蛋白的變化及運(yùn)動誘導(dǎo)HSP70對其影響,揭示運(yùn)動預(yù)防增齡性骨骼肌萎縮的作用機(jī)理。方法:27周齡雄性C57BL/6小鼠50只,經(jīng)1周適應(yīng)性喂養(yǎng)后,隨機(jī)分為年輕對照組(YS組)、老年對照組(OS組)和老年運(yùn)動組(OE組)。YS組(28周)小鼠經(jīng)抓力測試后立即處死取材。OE組小鼠采用30周、每周3~4次、每次50分鐘、75%VO2max有氧跑臺運(yùn)動訓(xùn)練(速度12米/分鐘,坡度0°)。OS組與OE組小鼠喂養(yǎng)至60周齡后,檢測抓力等指標(biāo)后處死。隨即分離各組小鼠腓腸肌,稱重后檢測小鼠骨骼肌萎縮指數(shù)(Sarcopenia index,SI),隨后提取腓腸肌總蛋白。采用Western Blot檢測小鼠腓腸肌組織HSP70,Cleaved Caspase-9蛋白表達(dá)水平;采用Co-IP法檢測小鼠腓腸肌HSP70與Apaf-1蛋白結(jié)合量;采用TUNEL法檢測小鼠腓腸肌細(xì)胞凋亡指數(shù)。結(jié)果:(1)SI變化:OS組小鼠腓腸肌SI較YS組顯著減少23.09%,而OE組SI與OS組相比顯著增加30.48%;(2)小鼠抓力:OS組小鼠抓力/體重比值比YS組顯著減少42.74%,OE組比OS組顯著增加21.51%;(3)凋亡指數(shù)檢測:伴隨年齡增加,OS組較YS組顯著增加475.28%,而有氧運(yùn)動之后,OE組小鼠凋亡指數(shù)較OS組顯著減少46.41%;(4)Western Blot結(jié)果:OS組Cleaved Caspase-9表達(dá)水平較YS組顯著增加45.40%,OE組較OS組顯著減少33.40%;(5)腓腸肌HSP70與Apaf-1蛋白的結(jié)合量:OS組較YS組HSP70與Apaf-1結(jié)合量顯著下降67.11%,運(yùn)動后OE組較OS組顯著增加306.31%。結(jié)論:(1)60周齡小鼠腓腸肌發(fā)生增齡性骨骼肌萎縮,30周有氧運(yùn)動可顯著延緩增齡性骨骼肌萎縮;(2)增齡性骨骼肌萎縮可能與骨骼肌凋亡細(xì)胞增多有關(guān),30周有氧運(yùn)動可顯著抑制骨骼肌細(xì)胞凋亡;(3)30周有氧運(yùn)動可增強(qiáng)HSP70與Apaf-1結(jié)合量,這可能是運(yùn)動抑制骨骼肌細(xì)胞凋亡,延緩增齡性骨骼肌萎縮的機(jī)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.
【作者單位】： 天津市天津醫(yī)院康復(fù)科;天津醫(yī)科大學(xué)基礎(chǔ)醫(yī)學(xué)院;天津體育學(xué)院運(yùn)動與健康科學(xué)系;
【基金】：天津市應(yīng)用基礎(chǔ)與前沿技術(shù)研究計(jì)劃項(xiàng)目(14JCYBJC27300) 國家自然科學(xué)基金(31571220)
【分類號】：R87

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