科罗索酸对胰岛素抵抗细胞糖代谢的影响及其机制研究

发布时间：2018-01-18 08:06

本文关键词：科罗索酸对胰岛素抵抗细胞糖代谢的影响及其机制研究　出处：《中南大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：研究科罗索酸对胰岛素抵抗3T3-1脂肪细胞和HepG2细胞糖代谢的影响,并对其机制进行初探。方法：采用高糖高胰岛素方法诱导3T3-L1脂肪细胞24小时建立胰岛素抵抗模型,用不同浓度的科罗索酸处理该模型细胞24小时,并设置正常对照、胰岛素抵抗模型组、二甲双胍阳性对照组,通过葡萄糖氧化酶法检测细胞培养基中葡萄糖的浓度来判断该药的疗效,提取细胞RNA进行RT-PCR实验,观察科罗索酸对脂肪细胞葡萄糖转运子4(GLUT4)、胰岛素受体底物1(IRS-1)、Cb1相关蛋白(CAP) mRNA表达的影响,并用HepG2细胞胰岛素抵抗模型验证科罗索酸的降糖效果。结果：科罗索酸和二甲双胍均能促进胰岛素抵抗3T3-L1脂肪细胞和HepG2细胞的糖代谢：1.在胰岛素抵抗3T3-L1脂肪细胞中,使用0.001、0.005、0.01、0.1、0.5、1mmol/L的科罗索酸处理的脂肪细胞的葡萄糖消耗量分别增加了5.60%、12.56%、22.82%、48.79%、27.80%、19.35%,0.1mmol/L的效果最强；经0.001、0.01、0.1、0.5、1mmol/L的二甲双胍处理的脂肪细胞的葡萄糖消耗量分别增加了2.44%、10.78%、28.49%、56.02%、55.04%,在0.5mmol/L时达最大值,之后趋于饱和；同等浓度的科罗索酸组与二甲双胍组比较,低浓度(0.001-0.1mmol/L)科罗索酸的效果优于二甲双胍,两者最大的葡萄糖消耗量均十分接近于正常水平,而科罗索酸所需的浓度仅为二甲双胍的1/5。与模型组比较,科罗索酸组的GLUT4和IRS-1基因的表达量分别增长了34.27%、22.05%,二甲双胍组的GLUT4和IRS-1基因的表达量分别增长了39.24%、28.57%。2.不同浓度(0.001、0.01、0.1、0.5、1mmol/L)的科罗索酸处理的HepG2细胞,其葡萄糖消耗量分别增加了0.7%、8.7%、16.5%、16.2%、11.0%。结论：科罗索酸能促进胰岛素抵抗脂肪细胞和HepG2细胞的糖代谢,减轻细胞的胰岛素抵抗；其降糖机制可能与GLUT4和IRS-1基因表达量的上调,促进相关蛋白的表达来调节葡萄糖的代谢。
[Abstract]:Aim: to study the effect of clofloxacin on glucose metabolism of insulin resistant 3T3-1 adipocytes and HepG2 cells and to explore its mechanism. Methods: 3T3-L1 adipocytes were induced with high glucose and high insulin for 24 hours to establish insulin resistance model. The model cells were treated with different concentrations of Corozo acid for 24 hours. Insulin resistance model group, metformin positive control group, glucose oxidase assay to determine the concentration of glucose in the cell culture medium to determine the efficacy of the drug, extract cell RNA for RT-PCR experiment. To observe the effect of Corozo acid on the mRNA expression of glucose transporter (GLUT4) and insulin receptor substrate (IRS-1) in adipocytes. HepG2 cell insulin resistance model was used to verify the hypoglycemic effect of Corozo acid. Results: both Corozo acid and metformin could promote glucose metabolism in insulin resistant 3T3-L1 adipocytes and HepG2 cells. Glucose consumption in adipocytes treated with 0. 001g / L Corozo acid increased by 5.60%, respectively. 12.56 with 22.82 and 48.79, with 27.80 and 19.350.The effect of 0.1 mmol / L was the strongest; The glucose consumption of adipocytes treated with 0.001mmol / L metformin increased by 2.44g 10.78%, respectively. 28.49 56.02 and 55.04, reaching the maximum at 0.5 mmol / L and then reaching saturation; In the same concentration of Corozo acid group compared with metformin group, the effect of Corozo acid at low concentration of 0.001-0.1 mmol / L was better than that of metformin group. The maximum glucose consumption of both groups was very close to the normal level, while the concentration of Corozo acid was only 1 / 5 of metformin, compared with the model group. The expression of GLUT4 and IRS-1 in Corozo acid group increased by 34.27% and 22.05% respectively. The expression of GLUT4 gene and IRS-1 gene in metformin group increased by 39.24 and 28.57, respectively. The glucose consumption of HepG2 cells treated with 1 mmol / L Corozo acid increased by 0.7 and 8.7 and 16.2and 11.0, respectively. Conclusion: Corozo acid can promote glucose metabolism of insulin resistant adipocytes and HepG2 cells and reduce insulin resistance. The hypoglycemic mechanism may be associated with the up-regulation of GLUT4 and IRS-1 gene expression and the regulation of glucose metabolism by promoting the expression of related proteins.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R96

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