本文選題：衰老 + 游泳運(yùn)動(dòng)��； 參考：《武漢體育學(xué)院》2017年碩士論文

【摘要】：目的:探究游泳運(yùn)動(dòng)干預(yù)對(duì)mi R-34a介導(dǎo)的細(xì)胞自噬在D-半乳糖誘導(dǎo)大鼠衰老進(jìn)程中的調(diào)控作用。方法:1.雄性Sprague-Dawley(SD)大鼠30只,隨機(jī)分為對(duì)照組(Control)、D-gal衰老模型組(D-gal)、衰老模型+運(yùn)動(dòng)干預(yù)組(D-gal+EXE),每組10只。一周的適應(yīng)性喂養(yǎng)后,D-gal+EXE組大鼠進(jìn)行適應(yīng)性游泳訓(xùn)練:第一天游泳10 min,之后每天增加10 min,直至增加到60 min。一周的適應(yīng)性訓(xùn)練后,開(kāi)始正式實(shí)驗(yàn)。每天對(duì)D-gal組和D-gal+EXE組大鼠按照200 mg/kg的劑量頸背部皮下注射D-半乳糖誘導(dǎo)衰老,同時(shí)D-gal+EXE組大鼠進(jìn)行60 min游泳訓(xùn)練,每周5天。藥物注射和游泳訓(xùn)練在均在每天同一時(shí)間進(jìn)行。6周的實(shí)驗(yàn)后,Morris水迷宮檢測(cè)各組大鼠學(xué)習(xí)記憶能力。水迷宮結(jié)束24小時(shí)后處死所有大鼠并取材。試劑盒檢測(cè)大鼠血清超氧化物歧化酶(SOD)活性及海馬組織中丙二醛(MDA)含量,Real-time-PCR檢測(cè)miR-34a的表達(dá)量,透射電鏡觀(guān)察海馬中線(xiàn)粒體形態(tài)變化及自噬小體,Western blot檢測(cè)細(xì)胞自噬及線(xiàn)粒體動(dòng)力學(xué)相關(guān)蛋白表達(dá)情況。2.培養(yǎng)SH-SY5Y細(xì)胞,當(dāng)細(xì)胞融合度達(dá)到50-70%時(shí)接種6孔板,轉(zhuǎn)染miR-34a抑制劑并用D-半乳糖刺激細(xì)胞,轉(zhuǎn)染后48 h收集細(xì)胞樣品,Real-time-PCR檢測(cè)miR-34a的表達(dá)情況,Western blot檢測(cè)細(xì)胞自噬及線(xiàn)粒體動(dòng)力學(xué)相關(guān)蛋白表達(dá)量。結(jié)果:1.Morris水迷宮結(jié)果顯示,與對(duì)照組相比,皮下注射D-半乳糖6周后,大鼠學(xué)習(xí)記憶能力明顯下降,而經(jīng)過(guò)6周游泳訓(xùn)練的D-gal+EXE組大鼠則有明顯改善。D-gal誘導(dǎo)的衰老大鼠穿越靶象限次數(shù)少于對(duì)照組大鼠,同時(shí)其潛伏期則高于對(duì)照組;而游泳運(yùn)動(dòng)干預(yù)組大鼠穿越靶象限次數(shù)明顯多于D-gal誘導(dǎo)模型組大鼠,其潛伏期則明顯短于衰老模型組。同時(shí)與對(duì)照組相比,D-gal模型組大鼠血清中SOD活性顯著下降,海馬組織中MDA含量顯著上升,運(yùn)動(dòng)干預(yù)后的D-gal+EXE組大鼠SOD則呈現(xiàn)顯著性提高,海馬中的MDA含量則呈現(xiàn)非常顯著性的下降。透射電鏡觀(guān)察發(fā)現(xiàn),D-gal模型組大鼠海馬中線(xiàn)粒體腫脹、融合增多,而D-gal+EXE組大鼠海馬組織中線(xiàn)粒體形態(tài)明顯趨于正常且有較多自噬小體。Real-time-PCR結(jié)果顯示:D-半乳糖誘導(dǎo)的衰老大鼠海馬組織中和D-半乳糖處理后的SH-SY5Y細(xì)胞中miR-34a的表達(dá)都顯著地升高,游泳干預(yù)和miR-34a抑制劑處理后其表達(dá)量展現(xiàn)為顯著性的下降。Western blot結(jié)果顯示,與正常對(duì)照組相比,D-gal模型組大鼠海馬中細(xì)胞自噬相關(guān)蛋白Atg7、Beclin1、LC3II/I均顯著下降,p62則顯著升高;線(xiàn)粒體動(dòng)力學(xué)相關(guān)蛋白Drp1和Mfn2表達(dá)量都顯著升高,D-gal+EXE組大鼠則Atg7、Beclin1、LC3II/I顯著升高,p62的表達(dá)量顯著下降;Drp1和Mfn2表達(dá)量也明顯下降。2.D-半乳糖處理后的SH-SY5Y細(xì)胞中自噬相關(guān)蛋白Atg7、Beclin1、LC3II/I均顯著下降,p62則顯著升高,線(xiàn)粒體動(dòng)力學(xué)相關(guān)蛋白Drp1和Mfn2表達(dá)量都顯著升高;mi R-34a抑制劑轉(zhuǎn)染后Atg7、Beclin1、LC3II/I顯著上升,p62的表達(dá)量顯著下降;Drp1和Mfn2表達(dá)量也都明顯下降。結(jié)論:在D-半乳糖誘導(dǎo)的大鼠腦衰老模型以及細(xì)胞衰老模型中,miR-34a通過(guò)調(diào)節(jié)細(xì)胞自噬以及線(xiàn)粒體動(dòng)力學(xué)相關(guān)蛋白的表達(dá)參與調(diào)控大鼠腦衰老進(jìn)程,游泳運(yùn)動(dòng)可以通過(guò)激活mi R-34a介導(dǎo)的細(xì)胞自噬改善線(xiàn)粒體功能,延緩腦衰老進(jìn)程。
[Abstract]:Objective: To explore the effect of swimming exercise intervention on MI R-34a mediated cell autophagy in the aging process of D- galactose induced rats. Methods: 1. male Sprague-Dawley (SD) rats were randomly divided into control group (Control), D-gal aging model group (D-gal), aging model + exercise intervention group (D-gal+EXE), 10 rats in each group. After the D-gal+EXE group, the rats of group D-gal+EXE were trained for adaptive swimming: swimming 10 min on the first day, and then increasing 10 min every day, until the adaptive training was increased to 60 min. a week, and the formal experiment began. Every day, the D-gal and D-gal+EXE group rats were injected with D- galactose under the neck of the neck of 200 mg/kg to induce aging, while the D-gal+EXE group was large. Rats were trained with 60 min swimming training, 5 days a week. After the drug injection and swimming training were performed at the same time of.6 weeks, the learning and memory ability of rats in each group was detected by the Morris water maze. After the water maze ended 24 hours, all rats were killed and selected. The serum superoxide dismutase (SOD) activity and hippocampus tissue of rats were detected by the reagent box. The content of MDA (MDA) and the expression of miR-34a were detected by Real-time-PCR. The morphologic changes of mitochondria in the hippocampus and autophagosomes in the hippocampus were observed by transmission electron microscope. Western blot was used to detect the expression of autophagy and mitochondrial dynamics related protein in.2. culture SH-SY5Y cells. When the degree of fusion reached 50-70%, the 6 pore plates were inoculated and the miR-34a inhibitors were transfected. The cells were stimulated with D- galactose, and the cells were collected at 48 h after transfection. The expression of miR-34a was detected by Real-time-PCR. Western blot was used to detect the expression of autophagy and mitochondrial dynamics related protein. Results: the results of 1.Morris water maze showed that the learning and memory ability of rats decreased significantly after 6 weeks of subcutaneous injection of D- galactose to the control group. The D-gal+EXE group after 6 weeks of swimming training had obviously improved.D-gal induced aging rats to cross the target quadrant times less than the control group, while the incubation period was higher than the control group, while the swimming exercise intervention group was more than the D-gal induced model group, and the incubation period was significantly shorter than that of the aging model rats. At the same time, compared with the control group, the activity of SOD in the serum of the D-gal model rats decreased significantly, the content of MDA in the hippocampus increased significantly. The SOD in the D-gal+EXE group of the group of D-gal+EXE rats showed significant improvement, and the content of MDA in the hippocampus decreased significantly. Transmission electron microscope observation found that the median grain of hippocampus in the D-gal model group was found in the rat model group. The body swelling and fusion increased, while the mitochondria in the hippocampus of D-gal+EXE rats tended to be normal and there were more autophagic bodies.Real-time-PCR results showed that the expression of miR-34a in the hippocampus of D- galactose induced aging rats and the SH-SY5Y cells treated with D- galactose were significantly increased, swimming intervention and miR-34a inhibitors were observed. After treatment, the expression amount showed a significant decrease in.Western blot results. Compared with the normal control group, the autophagy related protein Atg7, Beclin1, LC3II/I in the hippocampus of the D-gal model rats decreased significantly, and the p62 increased significantly; the mitochondrial kinetic related proteins Drp1 and Mfn2 increased significantly, and the D-gal+EXE group rats were Atg7, Beclin1, LC3II/I significantly increased, the expression of p62 decreased significantly, and the expression of Drp1 and Mfn2 also significantly decreased the autophagy associated protein Atg7, Beclin1, LC3II/I in SH-SY5Y cells after.2.D- galactose treatment. P62 was significantly increased, the mitochondrial kinetic related proteins Drp1 and expressions were significantly increased. TG7, Beclin1, LC3II/I significantly increased, the expression of p62 decreased significantly, and the expression of Drp1 and Mfn2 also decreased significantly. Conclusion: in the rat brain senescence model and cell aging model induced by D- galactose, miR-34a regulates the aging process of rat brain by regulating autophagy and the expression of mitochondrial dynamics related egg white. Activation can improve mitochondrial function and delay brain aging by activating mi R-34a mediated autophagy.
【學(xué)位授予單位】：武漢體育學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：G861.1;G804.2

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