替米沙坦对胰岛素抵抗OLETF大鼠心房肌L型钙通道α1C亚单位表达的影响

发布时间：2018-04-02 18:45

本文选题：替米沙坦　切入点：胰岛素抵抗　出处：《天津医科大学》2015年硕士论文

【摘要】：目的本实验以高脂饲料饲养自发性高血糖大鼠(otsuka long-evans tokushima fatty,OLETF),胰岛素抵抗(insulin resistance,IR)实验动物模型造模成功后,以替米沙坦进行药物干预,旨在:1、研究IR-OLETF大鼠心房肌L型钙通道α1C亚单位(L-type calcium channelα1C subunit,Cav1.2)的表达水平;2、研究血管紧张素II受体1型拮抗剂(angiotensin II type 1 receptor blocker,ARB)替米沙坦对IR-OLETF大鼠的心房肌Cav1.2蛋白表达的影响;3、探讨替米沙坦对心房肌Cav1.2蛋白表达的影响及其改善IR的可能机制。方法LETO(long-evans tokushima fatty)大鼠(雄性,1月龄,无特定病原体级)共计16只,OLETF大鼠(雄性,1月龄,无特定病原体级)共计32只,,体质量范围在135至185g,所有实验动物在标准饲料及干净整洁环境条件下适应性饲养1月。在2月龄时,所有OLETF大鼠以高脂饲料进行喂养,于7月龄时停止进食12小时后于股静脉处取血,测定所有实验动物的血清胰岛素(fasting insulin,FINS)、空腹血糖(fasting blood glucose,FBG)的数值,并通过相关公式分析得出胰岛素抵抗指数(insulin resistance index,HOMS-IR),分析大鼠胰岛素抵抗模型是否造模成功。用随机数字表法将32只造模成功的IR-OLETF大鼠分为2组,16只模型组(M)的大鼠继续以高脂饲料饲养,16只替米沙坦(T)组大鼠以高脂饲料饲养并加用替米沙坦(5mg/kgd)进行干预,16只LETO大鼠作为正常对照(NC)组,始终以标准饲料进行饲养。按上述方法饲养大鼠5月后,所用大鼠再次停止进食12小时后于股静脉处取血,通过放射免疫法检测FINS,以葡萄糖酶法检测FBG,使用全自动生化分析仪检测甘油三酯(triglyceride,TG)、高密度脂蛋白(high density lipoprotein cholesterol,HDL-C)、总胆固醇(total cholesterol,TC)及低密度脂蛋白(low density lipoprotein cholesterol,LDL-C)。然后使用脊椎脱臼法处死所用大鼠,迅速取出实验动物内脏(心脏、肝脏、肾脏、眼球等)组织保存于冻存管中,置入-80℃冰箱中低温密闭封存,使用为蛋白质印迹法检测所有实验动物心房肌组织中Cav1.2蛋白的表达量。采用SPSS20.0统计软件将所得结果进行计算分析。结果1、三组间实验大鼠的FINS,FBG,HOMA-IR的比较:T组和M组比较,FINS,FBG,HOMA-IR的数值明显降低,差异具有统计学意义;和NC组相比较,M组的FINS,FBG,HOMA-IR的数值显著增高,差异具有统计学意义(P0.05)。2、三组间实验大鼠血脂代谢水平的比较:T组和M组比较,LDL-C,TG,TC的数值显著降低,差异具有统计学意义(P0.05)。但HDL-C的数值无明显改变,差异无统计学意义(P=0.384);M组LDL-C,TG,TC的数值明显高于NC组,HDL-C数值明显低于NC组。3、三组间实验大鼠心房肌组织Cav1.2蛋白表达含量的比较:T组与M组相比,心房肌组织中Cav1.2蛋白水平明显升高,M组与NC组相比,心房肌组织中Cav1.2蛋白水平明显降低,差异有统计学意义(P0.05)。结论1、通过高脂饲养并成功建立胰岛素抵抗模型的OLETF大鼠血清中FINS,FBG,HOMA-IR,TG、LDL-C和TC数值显著升高,HDL-C数值显著降低。2、替米沙坦干预后,IR大鼠血清FBG,FINS,HOMA-IR,TC,TG和LDL-C水平明显降低,表明替米沙坦可以通过改善IR大鼠的糖脂代谢,改善其胰岛素抵抗。3、OLETF大鼠心房肌组织中Cav1.2的表达水平发现,胰岛素抵抗大鼠心房肌组织中Cav1.2蛋白的含量明显减少,而使用替米沙坦进行药物干预后其表达水平回升。
[Abstract]:The purpose of this experiment is fed with the high-fat diet in spontaneous hyperglycemic rats (Otsuka Long-Evans, Tokushima fatty, OLETF), insulin resistance (insulin resistance, IR) experimental animal model successfully, with telmisartan for drug intervention, aimed at: 1, atrial L research IR-OLETF rat calcium channel alpha subunit 1C (L-type calcium channel 1C subunit Cav1.2) alpha, the expression level; 2, study of angiotensin II type 1 receptor antagonist (angiotensin II type 1 receptor blocker, ARB) expression of telmisartan on IR-OLETF rat atrial muscle Cav1.2 protein; 3, to explore the possible mechanism of effect of telmisartan on the expression of Cav1.2 protein and atrial muscle improvement of IR method. LETO (Long-Evans Tokushima fatty) rats (male, 1 month old, specific pathogen free) a total of 16, OLETF rats (male, 1 month old, specific pathogen free), a total of 32, weight range In 135 to 185g, all the experiments in the standard animal feed and clean environment. In January 2 month old adaptive feeding, all OLETF rats were fed with high-fat diet, in 7 month old when to stop eating 12 hours after the blood samples were collected from the femoral vein, serum insulin were measured in all experimental animal (fasting insulin FINS) fasting blood glucose (fasting blood, glucose, FBG) value, and the insulin resistance index by correlation analysis formula (insulin resistance index, HOMS-IR), analysis of rat model of insulin resistance is the successful model. By the method of random number table 32 successful model of IR-OLETF rats were divided into 2 groups, 16 rats group (M) to rats by high fat diet, 16 telmisartan (T) group of rats fed with the high-fat diet combined with telmisartan (5mg/kg? D) intervention, 16 LETO rats as normal control group (NC), always with a standard diet According to the above method for feeding. Rats after May, rats used again to stop eating 12 hours after the blood samples were collected from the femoral vein, detected by FINS radioimmunoassay to detect FBG glucose enzymatic method, using automatic biochemical analyzer to detect triglyceride (triglyceride, TG), high density lipoprotein (high density lipoprotein cholesterol, HDL-C), total cholesterol (total, cholesterol, TC) and low density lipoprotein (low density lipoprotein cholesterol, LDL-C). Then use the spinal dislocation method executed by rats, quickly remove the experimental animal organs (heart, liver, kidney, eye) tissue preserved in freezing tube, using -80 C low temperature refrigerator sealed, used for Western blotting to detect the expression of Cav1.2 protein in all experimental animal atrial tissue in quantity. Using statistical software SPSS20.0 the results were calculated and analyzed. Results of the 1, between the three groups. Experimental rats FINS, FBG, HOMA-IR: T, group M and group FINS, FBG, HOMA-IR were significantly decreased, the difference was statistically significant; and NC group compared with M group, FINS, FBG, HOMA-IR values were significantly higher, the difference was statistically significant (P0.05.2), comparative experiment the lipid metabolism in rats: a comparison between the three groups, T group and M group LDL-C, TG, TC values decreased significantly, the difference was statistically significant (P0.05). But no significant changes in the value of HDL-C, the difference was not statistically significant (P=0.384); group M LDL-C, TG, TC value was significantly higher than NC group HDL-C numerical, significantly lower than NC group.3, between the three groups the expression of Cav1.2 protein in atrial tissue were compared in rats: compared with T group and M group, Cav1.2 protein levels in atrial tissue were significantly higher in M group compared with NC group, Cav1.2 protein levels in atrial tissue were significantly decreased, the difference was statistically significant (P0.05). Conclusion: 1, through High fat diet and successfully established FINS, OLETF in serum of rat model of insulin resistance in FBG, HOMA-IR, TG, LDL-C and TC values were significantly increased, HDL-C value decreased significantly.2 telmisartan intervention, serum FBG, rat IR FINS, HOMA-IR, TC, TG and LDL-C were significantly decreased, that of telmisartan can glycolipid improve the metabolism of IR rats, improve the insulin resistance of.3, the expression level of Cav1.2 in atrial tissue in OLETF rats found that insulin resistance Cav1.2 protein content of atrial tissue in rats was significantly reduced, and the use of drug intervention of telmisartan and its expression level rise.

【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R589.2

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