Roux-en-Y胃旁路术对肥胖症大鼠肾脏糖异生影响的实验研究

发布时间：2018-06-12 03:00

本文选题：肥胖症 + Roux-en-Y胃旁路术　；参考：《重庆医科大学》2017年硕士论文

【摘要】：目的:研究Roux-en-Y胃旁路术(Roux-en-Y gastric bypass,RYGB)对肥胖症大鼠糖代谢和肾脏糖异生的影响,初步探讨胃旁路术后肥胖症大鼠糖代谢和肾脏糖异生变化的原因,完善胃旁路术治疗肥胖症及其相关代谢紊乱的机制。方法:100只SD大鼠分别用高低脂饮食诱导构建90只高脂肥胖症大鼠和10只低脂对照大鼠,16周后将大鼠随机分为4组:对照组(低脂饮食诱导,n=8)、肥胖模型组(高脂饮食诱导,n=8)、假手术组(假手术干预肥胖模型组,n=8)和手术组(胃旁路术干预肥胖模型组,n=8),术后8周处死大鼠留取血清及组织标本。每周检测体质量及血糖值;术后血脂检测试剂盒测定各组大鼠血清总胆固醇(TG)、甘油三酯(TC)、低密度脂蛋白(LDL)和高密度脂蛋白(HDL)的浓度;术后酶联免疫吸附实验(ELASA)检测血清胰高血糖素样肽(GLP-1)和胰岛素(FINS)浓度后计算稳态模型的胰岛素抵抗指数(HOMA-IR),术前术后各组大鼠分别行腹腔内注射葡萄糖耐量实验(GTT)和腹腔内注射胰岛素耐量实验(ITT),梯形法计算血糖浓度-时间曲线下面积(AUG);术后骨密度仪(DEXA)成像、HE染色和免疫组化法分别观察大鼠体脂肪分布、组织形态学变化和糖异生关键酶定位;术后实时荧光定量聚合酶链式反应(RT-PCR)检测各组大鼠肾皮质糖异生关键酶,葡萄糖6磷酸酶(G6PC)和磷酸烯醇式丙酮酸羧激酶(PEPCK)m RNA的相对表达量;蛋白免疫印迹法(Western blot)检测各组大鼠肾皮质G6PC、PEPCK、IRS1和Akt及磷酸化蛋白的表达量。结果:1、体质量及血糖变化:术前相较于对照组,肥胖模型组大鼠体质量、血糖均明显升高(P0.05);肥胖模型组大鼠空腹血糖、GTT和ITT2小时内各时间点的血糖及AUG值较对照组明显升高(P0.05)。术后相较于肥胖模型组及假手术组,手术组大鼠体质量、血糖均明显降低(P0.05);手术组大鼠空腹血糖、GTT和ITT2小时内各时间点的血糖及AUG值较肥胖模型组及假手术组均明显降低。(P0.05)2、血生化及病理变化:相较于肥胖模型组及假手术组,手术组大鼠血清总胆固醇、甘油三酯及低密度脂蛋白浓度均明显降低(P0.05)。HE染色光镜下显示肥胖模型组和假手术组大鼠肾组织结构异常。手术组大鼠的肾脏组织病理损伤明显改善。相较于肥胖模型组及假手术组,手术组大鼠空腹血糖(FBG)、胰岛素(FINS)和HOMA-IR值均明显降低(P0.05),胰高血糖素样肽(GLP-1)水平显著升高(P0.05)。3、m RNA及蛋白表达水平相较于对照组,肥胖模型组及假手术组大鼠G6PC和PEPCK m RNA表达及蛋白含量均明显升高(P0.05),p-IRS1和p-Akt蛋白含量均明显降低(P0.05);手术组大鼠G6PC和PEPCK m RNA表达及蛋白含量较肥胖模型组及假手术组均明显降低(P0.05),p-IRS1和p-Akt蛋白含量明显升高(P0.05)。结论:1、高脂饮食诱导SD大鼠构建的肥胖症大鼠模型出现体质量升高、血糖升高、体脂肪百分比增加、血脂代谢紊乱、糖耐量受损及胰岛素抵抗,表现出肥胖症及相关代谢性紊乱的特点。2、Roux-en-Y胃旁路术后显著减轻肥胖症大鼠体质量、降低体脂肪百分比、改善血脂代谢紊乱和改善肾脏组织结构异常病理改变。3、Roux-en-Y胃旁路术后肥胖症大鼠血糖显著减低、糖耐量明显改善、胰岛素敏感性增加和肾皮质胰岛素抵抗现象显著缓解。胰岛素通路因子表达升高,肾皮质糖异生关键酶水平明显减低,肾脏糖异生作用被削弱,肾脏的糖输出减少,抑制内源性葡萄糖的产生。胃旁路术后糖脂代谢的改善与此密不可分。
[Abstract]:Objective: To study the effects of Roux-en-Y Roux-en-Y gastric bypass (RYGB) on glucose metabolism and kidney isogenesis in obese rats. The causes of carbohydrate metabolism and renal glucose isogenesis in obese rats after gastric bypass were preliminarily investigated. The mechanism of gastric bypass surgery for obesity and its related metabolic disorders was perfected. Methods: 100 SD rats respectively. 90 high fat obesity rats and 10 low fat control rats were induced with high and low fat diet. After 16 weeks, the rats were randomly divided into 4 groups: control group (low fat diet induction, n=8), obesity model group (high fat diet induction, n=8), sham operation group (sham operation intervention obesity model group, n=8) and operation group (n=8), 8 after operation, 8 after operation. The body mass and blood glucose were measured weekly. The serum serum total cholesterol (TG), triglyceride (TC), low density lipoprotein (LDL) and high density lipoprotein (HDL) were measured by the blood lipid detection kit after the operation. The serum glucagon like peptide (GLP-1) was detected by enzyme linked immunosorbent assay (ELASA) after operation. After the insulin (FINS) concentration, the insulin resistance index (HOMA-IR) was calculated in the steady state model. The rats before and after the operation were injected intraperitoneally with glucose tolerance test (GTT) and intraperitoneal injection of insulin tolerance test (ITT), and trapezoid method was used to calculate the area of blood glucose under time curve (AUG); postoperative bone densitometer (DEXA) imaging, HE staining and exemption The distribution of body fat, the histomorphology and the key enzyme location of the glucose isoseme were observed by the immunofluorescence method, and the relative expression of glucose 6 phosphatase (G6PC) and phosphoenolpyruvate carboxylic kinase (PEPCK) m RNA were detected by real time fluorescence quantitative polymerase chain reaction (RT-PCR). Western blot was used to detect the expression of G6PC, PEPCK, IRS1, Akt and phosphorylated protein in the renal cortex of rats. Results: 1, body mass and blood glucose change: the body mass and blood sugar of the obese model group were significantly increased before the operation compared with the control group (P0.05); the blood glucose and A of the rats in the obese model group were empty, GTT and ITT2 hours. The UG value was significantly higher than that in the control group (P0.05). Compared with the obesity model group and the sham operation group, the body mass and blood sugar of the operation group were significantly decreased (P0.05). The blood glucose and AUG values at all time points in the operation group were significantly lower than those in the obesity model group and the sham operation group. (P0.05) 2, the blood biochemical and pathological changes were: Compared to the obesity model group and the sham operation group, the concentration of serum total cholesterol, triglyceride and low density lipoprotein in the operation group decreased significantly (P0.05).HE staining light microscope showed that the renal tissue structure of the obese and sham operated rats was abnormal. The pathological damage of the kidney in the operation group was obviously improved. In the sham operation group, the fasting blood glucose (FBG), insulin (FINS) and HOMA-IR were significantly decreased (P0.05), and the level of glucagon like peptide (GLP-1) increased significantly (P0.05).3, m RNA and protein expression level was compared with the control group. The expression and protein content of G6PC and PEPCK m in the obesity model group and the sham operation group increased significantly. The content of RS1 and p-Akt protein decreased significantly (P0.05), and the expression and protein content of G6PC and PEPCK m RNA in the operation group were significantly lower than those in the obese model group and the sham operation group (P0.05), and the content of p-IRS1 and p-Akt proteins increased significantly (P0.05). Conclusion: 1, the high fat diet induced the obesity rat model of rats induced by the high fat diet to increase the body mass and increase the blood sugar. High percentage of body fat, dyslipidemia, impaired glucose tolerance and insulin resistance, showing the characteristics of obesity and related metabolic disorders.2, Roux-en-Y after gastric bypass significantly alleviated the body mass of obese rats, reduced the percentage of body fat, improved blood lipid metabolism disorder and improved renal tissue structure abnormal pathological changes.3, Roux- After en-Y gastric bypass surgery, the blood sugar of the obese rats was significantly reduced, the glucose tolerance was obviously improved, the insulin sensitivity was increased and the renal cortical insulin resistance was significantly relieved. The expression of insulin pathway factor increased, the level of the key enzymes in the renal cortex decreased obviously, the renal isogenic effect was weakened, the kidney sugar output decreased and the endogenous Portuguese was inhibited. The production of fructose is closely related to the improvement of glucose and lipid metabolism after gastric bypass surgery.
【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】：R656.6

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