运动对去卵巢大鼠骨骼肌芳香化酶及其相关信号通路蛋白的影响

发布时间：2018-05-15 10:37

本文选题：芳香化酶 + 大鼠　；参考：《上海体育学院》2017年硕士论文

【摘要】：研究目的:增龄性骨骼肌丢失问题已成为影响女性健康的重要因素之一。已有研究发现骨骼肌存在雌激素合成的关键酶——芳香化酶,而雌激素对骨骼肌有重要作用。运动对骨骼肌健康具有积极意义,但是运动能否促进骨骼肌芳香化酶的表达和雌激素的生成,以及局部合成的雌激素是否在运动的过程中发挥了改善肌肉功能的作用,相关研究尚未报道。本研究拟通过去卵巢动物模型探讨运动对骨骼肌芳香化酶表达的影响及其相关蛋白信号的表达,为运动改善低雌激素条件下骨骼肌质量提供理论基础和实验依据。研究方法:清洁级雌性SD大鼠25只,约12周龄,体重285.27±20.41g(由上海第二军医大学实验动物中心提供),适应性饲养1周后进行双侧卵巢切除手术。术后恢复2周进行分组。由于疾病等原因剔除5只,随机分为运动组(Trained组,10只)和安静组(Sedentary组,10只)。运动组进行8周有氧跑台训练,每周6次,25m/min;安静组正常饲养8周;干预结束后,取双侧比目鱼肌和跖肌。免疫荧光法检测右侧比目鱼肌和跖肌雌二醇和芳香化酶的水平。Western blotting技术检测MyoD、AKT、Fox O1/FoxO3、芳香化酶蛋白表达及磷酸化水平。研究结果:1、实验结束后两组大鼠体重无显著性差异(357.42±28.78g,347.14±31.46g),运动组跖肌和比目鱼肌质量均显著高于对照组(跖肌,459.28±32.15mg,396.78±26.46mg,p0.05;比目鱼肌,322.64±20.87mg,265.33±22.17mg,p0.05)。2、免疫荧光染色结果显示,实验结束后,运动组骨骼肌Aromatase阳性表达显著高于对照组(p0.05)。蛋白印迹分析结果显示运动组芳香化酶水平显著高于对照组(p0.01)。3、雌激素免疫荧光染色结果显示,运动组比目鱼肌雌二醇表达显著高于对照组(p0.05),而跖肌雌二醇表达水平运动组与对照组相比无显著性差异(p0.05)。4、蛋白印迹结果显示运动组跖肌和比目鱼肌MyoD、AKT的表达显著高于对照组,FoxO1/FoxO3的表达运动组显著低于对照组(p0.05)。研究结论:运动改善低雌激素水平下骨骼肌的质量,并且促进了骨骼肌芳香化酶表达和雌激素的生成。初步推断运动改善骨骼肌质量,可能与运动促进了雌激素的生成,进而调控了骨骼肌蛋白相关合成信号的表达有关。
[Abstract]:Objective: aging skeletal muscle loss has become one of the important factors affecting women's health. It has been found that aromatase, the key enzyme of estrogen synthesis, exists in skeletal muscle, and estrogen plays an important role in skeletal muscle. Exercise has a positive effect on skeletal muscle health, but whether exercise can promote the expression of aromatase and the production of estrogen in skeletal muscle, and whether the locally synthesized estrogen plays a role in improving muscle function during exercise. Related studies have not been reported. The purpose of this study was to investigate the effect of exercise on aromatase expression in skeletal muscle and the expression of related protein signal in ovariectomized animal model, and to provide theoretical and experimental basis for improving skeletal muscle quality under low estrogen condition. Methods: Twenty-five female SD rats of clean grade, aged about 12 weeks, weighing 285.27 卤20.41 g (supplied by Experimental Animal Center, Shanghai second military Medical University), were fed adaptively for one week and underwent bilateral ovariectomies. The patients were divided into groups 2 weeks after operation. For diseases and other reasons, 5 rats were randomly divided into exercise group (n = 10) and quiet group (n = 10). The exercise group received 8 weeks aerobic treadmill training six times a week for 25 m / min and the quiet group was fed for 8 weeks. After the intervention the soleus and plantar muscles were taken. The levels of estradiol and aromatase in the right soleus and plantar muscles were detected by immunofluorescence. The expression and phosphorylation of aromatase protein were detected by Western blotting. The results showed that there was no significant difference in body weight between the two groups after the end of the experiment. The weight of plantar muscle and soleus muscle in exercise group were significantly higher than those in control group (459.28 卤32.15 mg / g, 396.78 卤26.46 mg / g, 322.64 卤20.87 mg / L, 265.33 卤22.17 mg / g, P 0.05, respectively). The results of immunofluorescence staining showed that after the experiment, the weight of plantar muscle and soleus muscle in exercise group were significantly higher than those in control group (459.28 卤32.15 mg / g, 396.78 卤26.46 mg / g p 0.05; 322.64 卤20.87 mg / g, 265.33 卤22.17 mg / g, P 0.05, respectively). The positive expression of Aromatase in skeletal muscle in exercise group was significantly higher than that in control group (P 0.05). The results of Western blot analysis showed that the level of aromatase in exercise group was significantly higher than that in control group (P 0.01). The results of estrogen immunofluorescence staining showed that aromatase in exercise group was significantly higher than that in control group. The expression of estradiol in soleus muscle in exercise group was significantly higher than that in control group, while the level of estradiol expression in metatarsal muscle in exercise group was not significantly different from that in control group. Western blot analysis showed that MyoDfU AKT expression in metatarsal muscle and soleus muscle in exercise group was significantly higher than that in control group. The expression of Foxo 1 / FoxO 3 in exercise group was significantly lower than that in control group (P 0.05). Conclusion: exercise improves the quality of skeletal muscle at low estrogen level and promotes the expression of aromatase and the production of estrogen in skeletal muscle. It is inferred that exercise can improve the quality of skeletal muscle, which may be related to the regulation of estrogen production by exercise and the regulation of the expression of protein related synthesis signal in skeletal muscle.
【学位授予单位】：上海体育学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：G804.2

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