抗阻训练对胰岛素抵抗大鼠骨骼肌细胞外基质的影响

发布时间：2018-05-04 18:36

本文选题：胰岛素抵抗 + 细胞外基质　；参考：《上海体育学院》2017年硕士论文

【摘要】：研究目的通过高脂饲料喂养构建大鼠胰岛抵抗模型,通过负重爬梯抗阻训练,观察骨骼肌细胞外基质在胰岛素抵抗病理过程中的重塑,并进一步分析抗阻训练对骨骼肌细胞外基质病变的影响,为进一步探究抗阻训练改善胰岛素抵抗代谢健康提供方向和证据。研究方法30只SPF级SD大鼠随机分为对照组(C组)10只和高脂饲料喂养组(HF组)20只。通过高脂饲料喂养构建大鼠胰岛素抵抗模型,通过口服葡萄糖耐量实验(OGTT)并计算葡萄糖曲线下面积(AUC)来判定大鼠胰岛素敏感性,以C组与HF组AUC之间出现显著性差异(P0.05)为建模判定标准。建模成功后,随机从HF组选取10只大鼠组成抗阻训练组(HF+RT组)进行负重爬梯训练。负荷由初始体重的30%逐渐增加到120%,每周训练3次、每次3组、每组5个循环,干预13周。干预结束48h后测定大鼠胰岛素敏感性。大鼠麻醉后腹主静脉取血,检测血浆血脂四项、甘油三脂、游离脂肪酸和胰岛素。用免疫组化方法检测大鼠趾长伸肌细胞外基质胶原蛋白亚型colI、colIII、colIV的含量。用RT-PCR检测大鼠趾长伸肌炎性因子IL-6、IL-8、MCP-1、TGF-β、TNF-α的表达水平。用PCR array Rat Extracellular MatrixAdhesion Molecules检测趾长伸肌细胞外基质基因表达差异。用Western blot方法验证差异基因在蛋白水平的变化。研究结果1、建模HF组大鼠高脂进食3周后进食量明显小于C组(P0.05),但能量摄入始终高于C组(P0.05),并且体重在3周后开始明显高于C组(P0.05)。10周后,HF组葡萄糖耐量实验葡萄糖AUC值明显高于C组(P0.05),表明高脂饲料诱导机体胰岛素抵抗形成。2、体重、血脂与胰岛素敏感性HF组体重始终高于C组(P0.05),运动干预两周后,HF+RT组大鼠体重开始明显小于HF组(P0.05),并且与C组之间的差异失去显著性。与C组相比,HF组大鼠低密度脂蛋白胆固醇(LDL)和胰岛素水平升高(P0.05)。与HF组相比,HF+RT组总胆固醇(TC)、高密度脂蛋白胆固醇(HDL)、LDL和胰岛素的水平均下降(P0.05)。HF组葡萄糖耐量实验葡萄糖AUC值高于C组(P0.05),而HF+RT组AUC值介于C组和HF组,但与两组的差异均没有统计学意义(P0.05)。3、炎症指标在高脂诱导胰岛素抵抗的病理过程中,HF组IL-8 mRNA的水平显著升高,IL-6、MCP-1、TNF-α虽有升高趋势但没有统计学差异(P0.05)。与HF组相比,HF+RT组IL-6、IL-8、MCP-1、TNF-αmRNA均有下降的趋势,但没有统计学意义(P0.05)。4、胶原蛋白含量与C组相比,HF组colI、colIII、colIV蛋白含量都有显著性升高(p0.05)。抗阻训练可部分逆转这种改变:与HF组相比,HF+RT组colI、colIII有显著下降(P0.05),但仍高于C组(P0.05);colIV下降显著,达到与C组类似的水平(P0.05)。5、PCR array实验结果高脂膳食和高脂膳食合并抗阻训练引起骨骼肌ECM相关基因发生系类特异性的改变。与C组相比,HF组大鼠部分ECM相关基因mRNA水平显著升高,包括Cdh1、Col4a1、Col4a2、Itgal、Lamb3、Sele、Sparc和Thbs2(P0.05);HF+RT组Cdh1、Itgal、Sparc、Cdh3、Timp3、Sgce、Vtn、Actb mRNA水平显著升高(P0.05),而Sele和selp mRNA水平显著下降(P0.05)。部分基因的改变体现了抗阻训练对抗高脂膳食的作用,包括Itgal、Sele和Thbs2;与HF组相比,HF+RT组该类基因表达显著下降(P0.05)。6、Western blot实验结果根据PCR array实验结果,选取差异基因Itgal,Sele和Thbs2,检测其蛋白水平变化。与基因检测结果相吻合,与C组相比,Itgal、Sele和Thbs2蛋白水平均显著上升(P0.05);与HF组相比,抗阻训练可以显著降低Itgal、Sele和Thbs2蛋白水平表达(P0.05),以致Sele和Thbs2与C组差异消失(P0.05)。研究结论抗阻训练可以提高胰岛素抵抗大鼠的胰岛素敏感性,同时改善血脂代谢水平和骨骼肌炎性状态。抗阻训练在改善大鼠胰岛素抵抗的同时,改善了骨骼肌细胞外基质性变,包括降低骨骼肌ECM胶原蛋白沉积,降低ECM相关因子Itgal,Sele和Thbs2的基因表达和蛋白水平。
[Abstract]:The aim of this study is to construct rat islet resistance model by feeding high fat feed, and to observe the remodeling of the extracellular matrix of skeletal muscle during the pathological process of insulin resistance through the resistance training of weight climbing ladder, and to further analyze the effect of resistance training on the extracellular matrix lesion of skeletal muscle, and to further explore the resistance training to improve the insulin resistance generation. Xie Jiankang provided direction and evidence. Methods 30 SPF grade SD rats were randomly divided into 10 rats in the control group (group C) and 20 of the high fat feed group (group HF). The rat insulin resistance model was constructed by high fat feed, and the insulin sensitivity was determined by oral glucose tolerance test (OGTT) and the area of glucose curve (AUC) was calculated to determine the insulin sensitivity of rats. The significant difference (P0.05) between group C and group HF (P0.05) was established as a modeling criterion. After the success of the modeling, 10 rats were randomly selected from the HF group to form an anti resistance training group (HF+RT group) to carry out the staircase training. The load increased gradually from the initial weight to 120%, 3 times a week, 3 groups each time, 5 cycles in each group, and intervention for 13 weeks. The intervention ended 48h. After anesthesia, the rat insulin sensitivity was measured. The blood of the abdominal main vein was taken after anesthesia in rats. The blood lipid four items, glycerin three fat, free fatty acid and insulin were detected. The content of collagen subtype colI, colIII, colIV in the extracellular matrix of the extensor digitorum longus was detected by immunohistochemistry. RT-PCR was used to detect the IL-6, IL-8, MCP-1 of the extensitis factor of the rat toes. The expression level of TGF- beta, TNF- alpha. Using PCR array Rat Extracellular MatrixAdhesion Molecules to detect the difference in the expression of the extracellular matrix gene of the extensor digitorum longus. The change of the protein level was verified by Western blot method. Results 1, the intake of high fat feeding rats in the modeling HF group was less than that of the C group, but the energy intake was less than that of the C group, but the intake of energy was less than that of the C group. It was always higher than group C (P0.05), and the weight of body weight was obviously higher than that of group C (P0.05).10 weeks after 3 weeks. The glucose AUC value of glucose tolerance test in group HF was significantly higher than that of C group (P0.05), indicating that high fat diet induced body insulin resistance to form.2, body weight, blood lipid and Shima Somin perceptual HF group was always higher than that of C group, two weeks after exercise intervention. The weight of the T group was significantly smaller than that of the HF group (P0.05), and the difference between the group C and the group C was not significant. Compared with the C group, the low density lipoprotein cholesterol (LDL) and the insulin level in the group HF were increased (P0.05). The total cholesterol (TC), the high-density lipoprotein cholesterol (HDL), the levels of insulin and the insulin in the HF+RT group were lower than those in the group HF. Glucose tolerance test was higher than that in group C (P0.05), while AUC value in group HF+RT was between C and HF groups, but there was no significant difference between the two groups (P0.05), but the level of IL-8 mRNA in the HF group increased significantly in the pathological process of high fat induced insulin resistance. P0.05. Compared with the HF group, the IL-6, IL-8, MCP-1, TNF- alpha mRNA in the HF+RT group had a declining trend, but there was no statistical significance (P0.05).4. The content of collagen protein was significantly higher than that in the C group. Decreased (P0.05), but still higher than group C (P0.05), colIV decreased significantly, reached a similar level to C group (P0.05).5, PCR array experiment results showed that high fat diet and high fat diet combined with resistance training caused specific changes in skeletal muscle ECM related genes. Ol4a1, Col4a2, Itgal, Lamb3, Sele, Sparc, and Thbs2 (P0.05); HF+RT group Cdh1, Itgal, Sparc, etc. The expression of gene expression was significantly decreased (P0.05).6, and the results of Western blot experiment were based on the results of PCR array experiment. The difference gene Itgal, Sele and Thbs2 were selected to detect the change of protein level. The results of the gene detection were consistent with the gene detection results. Compared with the C group, Itgal, Sele and protein levels were significantly increased. Gal, Sele and Thbs2 protein levels are expressed (P0.05) so that the difference between Sele and Thbs2 and C disappears (P0.05). Conclusion resistance training can improve insulin sensitivity in insulin resistant rats and improve blood lipid metabolism and skeletal myositis. Resistance training improves skeletal muscle cells while improving insulin resistance in rats. Extracellular matrix changes include decreased ECM collagen deposition in skeletal muscle, and reduced gene expression and protein levels of ECM related factors Itgal, Sele and Thbs2.

【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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