GDF11在骨骼肌胰岛素抵抗中的作用研究

发布时间：2018-03-28 03:11

本文选题：GDF11　切入点：骨骼肌　出处：《重庆医科大学》2017年硕士论文

【摘要】：目的:最近研究发现GDF11可使衰老骨骼肌、心肌及大脑年轻化。既往研究报道衰老的骨骼肌与胰岛素抵抗密切相关。但GDF11是否具有改善胰岛素抵抗的作用尚未见明确的报道。因此,本研究拟探讨GDF11对骨骼肌胰岛素抵抗的作用。方法:体内实验采用低脂饮食、高脂饮食喂养小鼠,设置对照组及建立肥胖伴有胰岛素抵抗小鼠模型。IPGTT和ITT试验分别检测小鼠糖耐量和胰岛素敏感性。ELISA法检测小鼠血清GDF11水平,Western blotting和实时定量PCR分别检测小鼠骨骼肌中GDF11蛋白和基因水平的表达。体外实验采用棕榈酸干预骨骼肌细胞建立体外胰岛素抵抗模型。实时定量PCR检测棕榈酸干预后骨骼肌细胞GDF11基因水平的表达。体外采用GDF11干预棕榈酸诱导的骨骼肌细胞,2NBDG实验检测细胞糖摄取,Western blotting和实时定量PCR检测GLUT4,IRS-1及PGC-1α的表达。结果:(1)体内实验:与低脂喂养组相比,高脂喂养组小鼠体重明显增加,糖耐量异常,胰岛素敏感性降低(P0.05);与低脂喂养组比较,高脂喂养组小鼠血清中GDF11和骨骼肌中GDF11的表达均明显降低(P0.05)。(2)体外实验:与对照组相比,棕榈酸干预后骨骼肌细胞GDF11的表达明显降低,骨骼肌细胞糖摄取及糖摄取相关基因GLUT4和IRS-1的表达也明显下降(P0.05);加入GDF11干预后,骨骼肌细胞糖摄取及GLUT4和IRS-1的表达无明显变化(P0.05)。(3)与对照组相比,棕榈酸干预后骨骼肌细胞PGC-1α表达明显被抑制(P0.05);加GDF11干预后PGC-1α表达无明显变化(P0.05)。结论:本研究首次发现在肥胖伴胰岛素抵抗模型小鼠的血清和骨骼肌中GDF11蛋白明显降低,在体外棕榈酸干预骨骼肌细胞后GDF11表达也明显减少。而GDF11干预棕榈酸诱导的骨骼肌细胞,其胰岛素抵抗没有明显改善。
[Abstract]:Objective: recent studies have found that GDF11 can make skeletal muscle aging. Myocardial and brain rejuvenation. Previous studies have reported that aging skeletal muscle is closely associated with insulin resistance. However, whether GDF11 can improve insulin resistance has not been clearly reported. The aim of this study was to investigate the effect of GDF11 on insulin resistance in skeletal muscle. Methods: mice were fed with low fat diet and high fat diet in vivo. Establishment of obese mice model with insulin resistance. IPGTT and ITT test were used to detect glucose tolerance, insulin sensitivity, Elisa and serum GDF11 level. Western blotting and real-time quantitative PCR were used to detect skeletal muscle, respectively. In vitro insulin resistance model was established by palmitic acid intervention in skeletal muscle cells. Real-time quantitative PCR was used to detect the expression of GDF11 gene in skeletal muscle cells after palmitic acid intervention. GDF11 was used to interfere with palmitic acid-induced skeletal muscle cell line 2NBDG to detect the expression of GLUT4, IRS-1 and PGC-1 伪, and to detect the expression of GLUT4, IRS-1 and PGC-1 伪 by real-time quantitative PCR. Results: compared with low fat feeding group, the expression of GLUT4, IRS-1 and PGC-1 伪 was detected by GDF11 in vivo. Compared with low fat feeding group, the expression of GDF11 in serum and GDF11 in skeletal muscle were significantly decreased in high fat feeding group (compared with control group): compared with control group, compared with control group, the expression of GDF11 in serum and GDF11 in skeletal muscle were significantly decreased in high fat feeding group (P < 0. 05, P 0. 05, P 0. 05, P 0. 05, P 0. 05, P 0. 05, P 0. 05, P 0. 05). After palmitic acid intervention, the expression of GDF11 in skeletal muscle cells was significantly decreased, and the expression of glucose uptake and glucose uptake related genes GLUT4 and IRS-1 in skeletal muscle cells was also significantly decreased after the addition of GDF11. There was no significant change in glucose uptake and the expression of GLUT4 and IRS-1 in skeletal muscle cells. The expression of PGC-1 伪 in skeletal muscle cells was significantly inhibited after palmitic acid intervention, but no significant change was found in PGC-1 伪 expression after GDF11 treatment. Conclusion: the expression of GDF11 protein in serum and skeletal muscle of obese mice with insulin resistance was significantly decreased in this study. After palmitic acid intervention in vitro, the expression of GDF11 in skeletal muscle cells was also significantly decreased, while GDF11 intervention in palmitic acid-induced skeletal muscle cells had no significant improvement in insulin resistance.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R587.1

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