【摘要】：研究目的:運(yùn)動訓(xùn)練能夠提高機(jī)體的抗氧化能力。Nrf2作為機(jī)體氧化還原平衡的主要調(diào)節(jié)因子。運(yùn)動訓(xùn)練是否會影響Nrf2及抗氧化蛋白表達(dá),進(jìn)而影響體內(nèi)氧化還原狀態(tài),目前還不清楚。本實(shí)驗(yàn)通過對野生小鼠和Nrf2敲除小鼠在安靜和運(yùn)動兩種情況下,骨骼肌細(xì)胞中Nrf2及抗氧化蛋白表達(dá)的檢測,探討運(yùn)動訓(xùn)練對骨骼肌中Nrf2與抗氧化蛋白表達(dá)的影響。研究方法:野生C57BL/6J小鼠和Nrf2敲除小鼠各20只,根據(jù)體重,兩種鼠分為2組,每組10只,雌雄各半,分別為安靜組和運(yùn)動組,共4組。運(yùn)動組進(jìn)行四周跑臺運(yùn)動,每周運(yùn)動6天,休息一天,每天運(yùn)動1h,速度12m/min。正式實(shí)驗(yàn)前三天,運(yùn)動組小鼠進(jìn)行20min適應(yīng)性的跑臺運(yùn)動。在整個(gè)運(yùn)動過程中未對動物使用任何電刺激。運(yùn)動組進(jìn)行最后一次運(yùn)動之后休息48小時(shí)后同安靜組同脫頸處死,取材。Western blot法測定骨骼肌胞漿蛋白中4-HNE、Nrf2及抗氧化蛋白含量;RT-PCR測量Nrf2調(diào)節(jié)的抗氧化基因的表達(dá)量;ROS試劑盒測量骨骼肌內(nèi)ROS含量。研究結(jié)果:1)在安靜情況下,野生鼠和Nrf2敲除小鼠相比,骨骼肌中ROS以及CAT、NQO1、GCLc、HO-1和SOD2這五個(gè)抗氧化蛋白沒有顯著性改變。2)四周運(yùn)動訓(xùn)練之后,Nrf2敲除運(yùn)動組的ROS和大部分抗氧化蛋白都要顯著低于野生運(yùn)動組。研究結(jié)論:運(yùn)動訓(xùn)練通過Nrf2促進(jìn)小鼠骨骼肌抗氧化基因以及蛋白的表達(dá)。
[Abstract]:Objective: exercise training can improve the antioxidant ability of the body. Nrf2 is the main regulatory factor of redox balance. It is not clear whether exercise training will affect the expression of Nrf2 and antioxidant protein, and then the redox state in vivo. The effects of exercise training on the expression of Nrf2 and antioxidant protein in skeletal muscle cells were investigated by detecting the expression of Nrf2 and antioxidant protein in skeletal muscle cells of wild mice and Nrf2 knockout mice. Methods: there were 20 wild C57BL/6J mice and 20 Nrf2 knockout mice. According to their body weight, two kinds of mice were divided into 2 groups, 10 rats in each group, half male and half female, divided into four groups: quiet group and exercise group. The exercise group ran around the platform, 6 days a week, a day off, 1 hour a day, at a speed of 12 m / min. Three days before the formal experiment, the mice in the exercise group underwent 20min adaptive treadmill exercise. No electrical stimulation was used during the whole exercise. The exercise group took a rest for 48 hours after the last exercise and was executed with the same neck removal as the rest group. Determination of 4-HNE-Nrf2 in cytosolic protein of skeletal muscle by Western blot method and determination of antioxidant protein content by RT-PCR the expression of antioxidant gene regulated by Nrf2 was measured by Ros kit. ROS content in skeletal muscle was measured by RT-PCR. Results: 1) in quiet conditions, wild mice were compared with Nrf2 knockout mice. There was no significant change in ROS and SOD2 in skeletal muscle. The ROS and most antioxidant proteins in NRF 2 knockout exercise group were significantly lower than those in wild exercise group. Conclusion: exercise training can promote the expression of antioxidant genes and proteins in skeletal muscle of mice by Nrf2.
【學(xué)位授予單位】：北京體育大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：G804.7

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