大鼠运动减肥过程中骨骼肌硫化氢含量及肌肉因子的变化

发布时间：2018-08-19 16:30

【摘要】：背景:目前,肥胖已成为影响社会发展及个体健康的主要公共卫生问题。对现代人来说,营养过剩及体力活动减少是引起或加重肥胖的关键因素,因此运动在预防和控制肥胖中占据不可替代的作用。骨骼肌不仅是参与运动的主要器官,而且具有重要的内分泌功能。骨骼肌在收缩过程中可分泌多种细胞因子,其中一些细胞因子除了在局部发挥作用外,还可分泌入血并进入其它组织和器官发挥其生理功能,这些细胞因子被称为肌肉因子。IL-6作为最早发现的肌肉因子,其与能量代谢及运动的关系已有不少报道,然而这些研究都是在正常健康状态下进行,而对病理状态下,如肥胖状态下运动对IL-6的调节作用还无人研究。IL-15是一种新确定的肌肉因子,它主要由骨骼肌产生,可促进骨骼肌蛋白的积累,抑制其分解;此外,IL-15还可作用于脂肪组织内的IL-15受体进而调节脂肪的代谢,因此对治疗肥胖及骨骼肌萎缩具有积极作用。有趣的是,运动对防治肥胖和骨骼肌萎缩也有重要作用,其机制是否与IL-15有关,目前还不清楚。硫化氢(H2S)是一种新近发现的的气体分子递质,其在代谢、炎症、神经和血管等多个方面所起到的调节作用已被越来越多的研究所证实。最近有文献报道,外源性H2S具有改善脂质代谢并减轻脂肪肝的作用。催化生成内源性H2S的三种酶在大鼠骨骼肌中均有表达,而运动对骨骼肌内源性H2S合成系统的影响还未见报道。目的:检测大鼠运动减肥过程中骨骼肌内源性H2S合成系统及IL-6、IL-15基因表达的变化,为深入理解运动减肥的机制及H2S在运动减肥中所起的作用提供依据。方法:以SD雄性大鼠为研究对象,采用8周造模法建立大鼠肥胖模型并用中等强度跑台运动对高脂膳食肥胖大鼠进行干预。实验共分三组:正常对照组(CN组)、肥胖组(OB组)、肥胖加运动组(OB+EX组)。用酶联免疫吸附实验(ELISA)检测各实验组大鼠骨骼肌中H2S的含量;用RT-PCR、Real-time PCR和Western blotting实验检测大鼠骨骼肌中H2S合成相关酶胱硫醚γ裂解酶(CSE)、胱硫醚β合成酶(CBS)及3-巯基丙酮酸硫基转移酶(MST)的m RNA和蛋白表达情况;用免疫组化(IHC)实验检测大鼠骨骼肌中CSE的定位及蛋白表达量;用RT-PCR检测骨骼肌中IL-6、IL-15的m RNA表达。结果:ELISA结果显示:与CN组相比,OB组大鼠骨骼肌中H2S含量明显降低(P0.01);与OB组相比,OB+EX组大鼠骨骼肌中H2S含量有所升高(P0.01)。Real-time PCR结果显示:与CN组相比,OB组大鼠骨骼肌中CSE基因的m RNA表达量明显下降(P0.01),CBS基因的m RNA表达量也降低(P0.05);与OB组相比,OB+EX组大鼠骨骼肌中CSE基因的m RNA表达量升高(P0.05)。Western blotting结果显示:与CN组相比,OB组大鼠骨骼肌中CSE基因的蛋白表达量明显下降(P0.01);与OB组相比,OB+EX组大鼠骨骼肌中CSE基因的蛋白表达量明显升高(P0.01)。IHC结果显示,大鼠骨骼肌中CSE蛋白主要定位于胞质内,运动组大鼠骨骼肌中CSE蛋白表达量明显升高。RT-PCR结果显示:与CN组相比,OB组大鼠骨骼肌中IL-6基因的m RNA表达量明显升高(P0.01),IL-15的m RNA表达量无变化;与OB组相比,OB+EX组大鼠骨骼肌中IL-6基因的m RNA表达量降低(P0.05),IL-15基因的m RNA表达量无明显差异。结论:1.肥胖降低大鼠骨骼肌中H2S的含量,而中等强度运动使其升高,这可能与内源性H2S合成酶CSE的规律性变化有关。2.肥胖升高大鼠骨骼肌中IL-6的m RNA表达量,而中等强度运动使其降低,这可能与运动升高骨骼肌中H2S的含量有关。
[Abstract]:BACKGROUND: Obesity has become a major public health problem affecting social development and individual health. For modern people, overnutrition and reduced physical activity are the key factors that cause or aggravate obesity, so exercise plays an irreplaceable role in preventing and controlling obesity. Skeletal muscle can secrete a variety of cytokines during contraction, some of which can not only play a role in the local, but also secrete into the blood and into other tissues and organs to play their physiological functions. These cytokines are known as muscle factors. IL-6 as the earliest discovered muscle factors, with There have been many reports on the relationship between energy metabolism and exercise. However, these studies have been carried out under normal health conditions. However, no one has studied the regulation of exercise on IL-6 in pathological conditions, such as obesity. Interestingly, exercise also plays an important role in the prevention and treatment of obesity and skeletal muscle atrophy. Whether the mechanism is related to IL-15 is unclear. Recently, it has been reported that exogenous H2S can improve lipid metabolism and reduce fatty liver. Three enzymes that catalyze the formation of endogenous H2S are expressed in skeletal muscle of rats. Objective: To investigate the changes of endogenous H2S synthesis system and the expression of IL-6 and IL-15 genes in skeletal muscle during exercise-induced weight loss in rats, so as to provide a basis for further understanding the mechanism of exercise-induced weight loss and the role of H2S in exercise-induced weight loss. The experiment was divided into three groups: normal control group (CN group), obesity group (OB group) and obesity plus exercise group (OB + EX group). The content of H2S in skeletal muscle was detected by enzyme linked immunosorbent assay (ELISA). R, Real-time PCR and Western blotting were used to detect the m RNA and protein expression of H2S synthase-related enzymes cystathionine gamma lyase (CSE), cystathionine beta synthase (CBS) and 3-mercaptopyruvate sulfotransferase (MST) in skeletal muscle of rats; immunohistochemistry (IHC) was used to detect the localization and protein expression of CSE in skeletal muscle of rats; RT-PCR was used to detect the expression of CSE. Results: Compared with CN group, the content of H2S in skeletal muscle of OB group was significantly lower (P 0.01); compared with OB group, the content of H2S in skeletal muscle of OB + EX group was higher (P 0.01). Real-time PCR showed that the expression of M RNA of CSE gene in skeletal muscle of OB group was significantly lower than that of CN group. Compared with OB+EX group, the expression of CSE gene in skeletal muscle of OB+EX group increased (P 0.05). The Western blotting results showed that compared with CN group, the expression of CSE gene in skeletal muscle of OB+EX group decreased significantly (P 0.01). The results of IHC showed that CSE protein was mainly localized in the cytoplasm of skeletal muscle of rats, and the expression of CSE protein in skeletal muscle of rats in exercise group was significantly increased. Compared with OB group, the expression of IL-6 m RNA in skeletal muscle of OB+EX group decreased (P 0.05), but the expression of IL-15 m RNA had no significant difference. Conclusion: 1. Obesity decreased the content of H2S in skeletal muscle of rats, while moderate intensity exercise increased it, which may be related to the regular changes of endogenous H2S synthase CSE. 2. Obesity increased the bone of rats. The expression of IL-6 m RNA in skeletal muscle was decreased by moderate intensity exercise, which may be related to the increase of H2S content in skeletal muscle.
【学位授予单位】：河南大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R589.2

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