本文選題：大負(fù)荷運(yùn)動(dòng) + 骨骼肌��； 參考：《北京體育大學(xué)》2016年博士論文

【摘要】：目的：觀察一次大負(fù)荷運(yùn)動(dòng)對(duì)大鼠骨骼肌線粒體結(jié)構(gòu)和功能的影響,分析線粒體自噬蛋白及相關(guān)蛋白的變化,探討大負(fù)荷運(yùn)動(dòng)誘導(dǎo)骨骼肌線粒體自噬的分子機(jī)制；觀察針刺對(duì)大負(fù)荷運(yùn)動(dòng)后骨骼肌線粒體的影響,并闡明其作用機(jī)理。方法：將雄性SD大鼠作為研究對(duì)象。研究一和研究二分為對(duì)照組(C)和運(yùn)動(dòng)組(E),E組進(jìn)行一次大負(fù)荷下坡跑；研究三分為安靜對(duì)照組(C)、單純運(yùn)動(dòng)組(E)、單純針刺組(A)和運(yùn)動(dòng)針刺組(EA),E和EA組進(jìn)行下坡跑運(yùn)動(dòng),A組和EA組施加針刺處理。各組又按時(shí)相點(diǎn)劃分為0h、12h、24h、48h和72h組,分別于對(duì)應(yīng)時(shí)間點(diǎn)分離大鼠比目魚肌進(jìn)行檢測(cè)。使用透射電鏡觀察骨骼肌線粒體超微結(jié)構(gòu)變化；采用ELISA方法測(cè)定線粒體定量酶CS的含量以及線粒體呼吸鏈復(fù)合體Ⅱ、Ⅳ的活性變化；應(yīng)用Western Blot方法檢測(cè)骨骼肌COX Ⅰ、PINK1和線粒體中Parkin、LC3的蛋白表達(dá)；利用免疫熒光雙標(biāo)技術(shù)觀測(cè)Parkin、Ub、p62、LC3分別與線粒體的共定位及量。結(jié)果：(1)一次大負(fù)荷運(yùn)動(dòng)后比目魚肌線粒體出現(xiàn)明顯腫脹、肌膜下積聚等超微結(jié)構(gòu)異常變化,且伴有大量不同成熟階段的自噬體形成,CS的含量減少,呼吸鏈復(fù)合體Ⅱ、Ⅳ活性以及COX Ⅰ蛋白表達(dá)出現(xiàn)一過性的下調(diào)。(2)一次大負(fù)荷運(yùn)動(dòng)后線粒體自噬蛋白PINKl、Parkin以及自噬相關(guān)蛋白Ub、p62、LC3均出現(xiàn)一過性的表達(dá)升高,免疫熒光結(jié)果和蛋白表達(dá)結(jié)果一致。(3)運(yùn)動(dòng)后針刺能明顯改善線粒體超微結(jié)構(gòu)的病理學(xué)改變,有自噬溶酶體的出現(xiàn),抑制CS的含量減少,上調(diào)呼吸鏈復(fù)合體Ⅱ、Ⅳ活性以及COX Ⅰ蛋白表達(dá),下調(diào)PINK1、Parkin、Ub、p62、LC3在線粒體上的表達(dá),免疫熒光結(jié)果和蛋白表達(dá)結(jié)果一致。結(jié)論：(1)一次大負(fù)荷運(yùn)動(dòng)可導(dǎo)致骨骼肌線粒體結(jié)構(gòu)和功能受損,有大量自噬體形成,可能誘導(dǎo)了線粒體自噬的發(fā)生。(2)一次大負(fù)荷運(yùn)動(dòng)激活PINK1/Parkin通路,進(jìn)而使Ub和p62在線粒體上表達(dá)最終促進(jìn)LC3與線粒體結(jié)合,三者協(xié)同調(diào)控線粒體自噬的發(fā)生。(3)運(yùn)動(dòng)后針刺使線粒體外膜蛋白PINK1表達(dá)下調(diào),減少對(duì)胞漿中Parkin的招募,進(jìn)而影響其下游的Ub和p62發(fā)生線粒體轉(zhuǎn)位及LC3與線粒體的結(jié)合,調(diào)節(jié)了自噬溶酶體通路并抑制了線粒體自噬過度激活,同時(shí)緩解了線粒體結(jié)構(gòu)和功能的破壞。
[Abstract]:Objective: to observe the effect of a heavy load exercise on the structure and function of mitochondria in skeletal muscle of rats, to analyze the changes of mitochondrial autophagy protein and related proteins, and to explore the molecular mechanism of mitochondrial autophagy induced by heavy exercise. To observe the effect of acupuncture on mitochondria of skeletal muscle after heavy exercise and to elucidate its mechanism. Methods: male SD rats were studied. The first and second studies were divided into two groups: control group (C) and exercise group (group E). The three groups were divided into three groups: silent control group, simple exercise group, simple acupuncture group, and exercise acupuncture group and EA group were treated with acupuncture in downhill running exercise group A and EA group. Each group was divided into two groups: 0 h, 12 h, 24 h, 48 h and 72 h, respectively. The soleus muscle was separated at the corresponding time point for detection. The ultrastructure of mitochondria in skeletal muscle was observed by transmission electron microscope (TEM), the content of mitochondrial quantitative enzyme CS and the activity of mitochondrial respiratory chain complex 鈪，

本文編號(hào)：1955427