二甲双胍对糖尿病大鼠FGF19，FGF21和FGFR1的影响

发布时间：2018-06-06 07:01

本文选题：二甲双胍 + 成纤维细胞生长因子19　；参考：《山东大学》2017年硕士论文

【摘要】：研究目的近年来研究发现成纤维细胞生长因子19(FGF19)、成纤维细胞生长因子21(FGF21)通过与成纤维细胞生长因子受体(FGFRs)结合在糖脂代谢中发挥了重要作用,被认为新的代谢激素。此外,研究表明特异性活化纤维细胞生长因子受体1(FGFR1)可以改善糖脂代谢。二甲双胍作为临床一线用药,主要通过抑制肝糖原输出和增加外周组织胰岛素敏感性发挥作用。我们研究二甲双胍干预前后2型糖尿病模型中血清FGF19、FGF21,胰腺和脂肪组织FGFR1的表达水平及Akt的磷酸化水平的变化,进而探讨二甲双胍新的药物靶点和器官靶点。研究方法给予8周龄健康雄性Wistar大鼠(n=60只)标准饮食,随机分为正常对照组(Control group,Con)(n=20)和高脂饮食组(high fat diet group,HFD)(n=40),分别普通饲料和高脂饲料喂养。6w后,将HFD组大鼠腹腔注射链脲佐菌素(STZ)(35mg/kg)诱导糖尿病模型。诱导成模的大鼠随机分为DM组和DM+METF组,DM+METF大鼠给予二甲双胍(500 mg·kg-1·-d-1)。在24周末检测血清生化学指标,酶联免疫吸附法(ELISA)检测FGF19和FGF21水平。蛋白免疫印迹(WB),免疫组化(IHC)检测组织FGFR1的表达水平。WB检测总的Akt(total Akt)及磷酸化的Akt(p-Akt)的变化。三组之间比较采用 one-way AN0VA。实验结果1.各组大鼠一般观察指标的比较糖尿病组(DM)大鼠在高脂饮食/STZ造模成功后,进食量和饮水量及体重显著增加,高于正常对照组。二甲双胍干预糖尿病组(DM+METF)大鼠的进食量,饮食量和体重也较正常组显著增加,但是均明显低于糖尿病组大鼠,差异具有统计学意义(P＜0.05)。2.各组大鼠的血清生化学指标比较在实验末,糖尿病组(DM)大鼠的糖脂和胆汁酸代谢紊乱。2型糖尿病大鼠的糖耐量实验提示胰岛素释放高峰延迟,整体胰岛素曲线低平,两小时末,胰岛素水平仍高于正常对照组。二甲双胍干预糖尿病组大鼠的FBG、FIN、TC、TG、LDL均较DM组大鼠降低,差异具有统计学意义。3.各组大鼠FGF19,FGF21,FGFR1和Akt磷酸化水平比较与正常对照组(Con)组大鼠比较,糖尿病组(DM)组大鼠血清FGF19水平显著降低(P＜0.05);FGF21水平显著升高(P＜0.05);脂肪组织和胰腺组织FGFR1表达异常增高,Akt的磷酸化水平(p-Akt/total-Akt)显著降低(P＜0.05)。二甲双胍干预大鼠的血清FGF21较DM组大鼠明显升高(P0.05),而FGF19较DM组大鼠明显减低(P0.05)。此外,胰腺FGFR1的表达水平接近正常组水平,而脂肪组织的FGFR1仍高于正常对照组和糖尿病组;两组织的Akt的磷酸化水平(p-Akt/total-Akt)明显高于糖尿病组大鼠。结论1.高脂饮食和小剂量链脲佐菌素(35mg/kg)可以诱导2型糖尿病模型,2型糖尿病大鼠的血脂和血糖代谢紊乱。2.二甲双胍可以通过抑制摄食量改善体重。此外,还可以使胆固醇向胆汁酸转化增多,并促进胆汁排泄。3.2型糖尿病大鼠FGF19和FGF21代谢出现紊乱,脂肪和胰腺组织FGFR1及Akt通路损伤。4.二甲双胍干预可以使得DM大鼠的FGF19进一步降低,FGF21升高。但是可以逆转Akt磷酸化水平的降低,这可能是通过FGFR1作用于Akt通路从而改善胰腺及脂肪组织功能。
[Abstract]:Research aims to find fibroblast growth factor 19 (FGF19), fibroblast growth factor 21 (FGF21), which plays an important role in glycolipid metabolism by combining with fibroblast growth factor receptor (FGFRs), and is considered as a new metabolic hormone. In addition, the study showed that specific activated fibroblast growth factor receptor 1 (FGFR). 1) can improve glycolipid metabolism. Metformin as a clinical first-line drug, mainly by inhibiting liver glycogen output and increasing peripheral tissue insulin sensitivity. We studied the expression level of serum FGF19, FGF21, pancreas and adipose tissue FGFR1 and the changes of phosphorylation level of Akt in type 2 diabetes mellitus model before and after metformin intervention. And then discuss the new drug target and organ target of metformin. The study method was given to 8 weeks old healthy male Wistar rats (n=60 only) standard diet and randomly divided into normal control group (Control group, Con) (n=20) and high fat diet group (high fat diet group, HFD) (n= 40). Streptozotocin (STZ) (35mg/kg) induced diabetes model was induced. The rats were randomly divided into DM group and DM+METF group. DM+METF rats were given metformin (500 mg. Kg-1. -d-1). Serum biochemical indexes were detected at the end of the 24 week, FGF19 and FGF21 levels were detected by enzyme linked immunosorbent assay (ELISA). Protein immunoblotting (WB) and immunohistochemical staining were detected. The expression level of FGFR1 was measured by.WB, the total Akt (total Akt) and phosphorylation of Akt (p-Akt). The comparison between the three groups was compared with the results of one-way AN0VA. experiment 1., the general observation index of rats in each group was compared with the diabetic group (DM) rats. After the high fat diet /STZ model was successful, the food intake and the amount of drinking water and body weight were significantly increased, higher than normal. Control group. Metformin intervention in diabetic group (DM+METF) rats intake, diet and weight was also significantly higher than the normal group, but all significantly lower than the diabetic rats, the difference was statistically significant (P < 0.05) the serum biochemical indexes of rats in.2. groups were compared at the end of the experiment, glucose and bile acid metabolism in diabetic group (DM) rats The glucose tolerance test of type.2 diabetic rats showed that the peak of insulin release was delayed, the overall insulin curve was low, and the insulin level was still higher than that of the normal control group at the end of two hours. The FBG, FIN, TC, TG, LDL of the diabetic rats were lower than those in the DM group, and the difference was statistically significant in FGF19, FGF21, FGFR1 in each group of.3. rats. Compared with the normal control group (Con), the level of serum FGF19 in the diabetic group (DM) was significantly lower (P < 0.05), the level of FGF21 increased significantly (P < 0.05), the expression of FGFR1 in the adipose tissue and pancreatic tissue was increased, and the phosphorylation level of Akt (P < 0.05) was significantly decreased (P < 0.05). The intervention of metformin in the DM group was significantly lower. The serum FGF21 of rats was significantly higher than that of the DM group (P0.05), while FGF19 was significantly lower than that in the DM group (P0.05). In addition, the expression level of the pancreas FGFR1 was close to that of the normal group, but the FGFR1 of the adipose tissue was still higher than that of the normal control group and the diabetic group; the phosphorylation level of Akt (p-Akt/total-Akt) in two tissues was significantly higher than that of the diabetic rats. Conclusion 1 High fat diet and small dose of streptozotocin (35mg/kg) can induce type 2 diabetes model. The blood lipid and blood glucose metabolism disorder.2. metformin in type 2 diabetic rats can improve the body weight by inhibiting the intake of food. In addition, it can also increase the conversion of cholesterol to bile acid, and promote the bile excretion of the FGF19 and FGF21 generation of type.3.2 diabetic rats. The metabolic disorder of FGFR1 and Akt pathway in fat and pancreatic tissue damage.4. metformin intervention can further reduce FGF19 in DM rats and increase FGF21. But it can reverse the decrease in the level of Akt phosphorylation, which may be through the action of FGFR1 on the Akt pathway to improve the function of the pancreas and fat tissue.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R587.1

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