利拉鲁肽与二甲双胍联用对糖尿病大鼠胰岛素抵抗、胸主动脉纤维化的影响

发布时间：2018-05-12 03:45

本文选题：糖尿病大鼠 + 利拉鲁肽　；参考：《皖南医学院》2017年硕士论文

【摘要】：目的:观察利拉鲁肽与二甲双胍联合用药对糖尿病大鼠血糖、血脂、胰岛素抵抗的影响;对胸主动脉纤维化的影响及其可能机制。方法:随机抽取SPF级SD雄性大鼠9只作为正常对照组(NG组,双蒸水,5 ml·kg-1·d-1,i.g.),其余大鼠采用高糖高脂饲养加腹腔注射链脲佐菌素(STZ)35mg·kg-1诱导糖尿病大鼠模型。将造模成功的大鼠(空腹血糖≥11.1 mmol/L)随机分为糖尿病模型对照组(DM组,双蒸水,5 ml·kg-1·d-1,i.g.),二甲双胍治疗组(Met组,100 mg·kg-1·d-1,i.g.),利拉鲁肽治疗组(Lir组,0.2 mg·kg-1·d-1,i.p.),利拉鲁肽联用二甲双胍治疗组(Lir+Met组,0.1 mg·kg-1·d-1,i.p.+50 mg·kg-1·d-1,i.g.),格列齐特联用二甲双胍治疗组(Gli+Met组,10 mg·kg-1·d-1+50 mg·kg-1·d-1,i.g.)。各组于每天早上8点-10点灌胃或腹腔注射给予相应药物,连续给药6周。期间每2周称重一次、检测大鼠空腹血糖(FBG)变化。于给药6周末进行口服糖耐量(OGTT)实验,计算曲线下面积(AUC)。腹主动脉取血,检测大鼠血脂水平、糖化血红蛋白(HbA1c)和空腹血清胰岛素(FINS)水平,并计算胰岛素抵抗指数(IR);测定血清中肿瘤坏死因子(TNF-α)、血管内皮生长因子(VEGF)、大鼠血管性血友病因子(VWF)含量;HE染色法观察大鼠胸主动脉病理损伤;Masson染色法观察大鼠胸主动脉胶原纤维变化;测算胶原容积(CVF);RT-PCR法检测TNF-α、VEGF、丝裂原活化蛋白激酶p38(P38MAPK)mRNA表达。结果:(1)与正常组相比较,糖尿病模型组大鼠体重(BW)、高密度脂蛋白胆固醇(HDL-C)均显著下降(P0.01),而FBG、FINS、TG、TC、LDL-C、HbA1c则显著上升(P0.01或P0.05),提示出现明显的胰岛素抵抗;血清中TNF-α、VEGF、VWF含量升高(P0.01或P0.05);HE染色显示胸主动脉内膜增厚,内皮细胞、平滑肌排列紊乱,炎性物质浸润,产生明显的病理损伤;Masson染色出现胶原纤维沉积,血管增厚,CVF显著升高(P0.01);RT-PCR法检测显示TNF-α、VEGF和P38MAPK mRNA表达升高(P0.01或P0.05)。(2)与糖尿病模型组大鼠相比较,各给药组大鼠BW、HDL-C有所上升(P0.01或P0.05),FBG、FINS、TG、LDL-C、HbA1c有不同程度的下降(P0.01或P0.05),HOMA-IR有所下降(P0.01或P0.05),且利拉鲁肽联合二甲双胍用药组效果优于二甲双胍单用药组;血清中TNF-α、VEGF、VWF有不同程度的降低(P0.01或P0.05),利拉鲁肽联合二甲双胍组降低效果显著(P0.01或P0.05);HE染色显示各给药组大鼠胸主动脉炎性浸润、内膜增生等有不同程度改善,病理损伤减轻,且利拉鲁肽联合二甲双胍用药组效果优于单用药组;Masson染色显示各给药组能减少胸主动脉胶原纤维沉积、血管增厚等现象,显著降低CVF(P0.01),利拉鲁肽联合二甲双胍降低CVF作用效果与联用药对照组相当;RT-PCR法显示各给药组TNF-α、VEGF和P38MAPK mRNA表达有所下降(P0.01或P0.05),而利拉鲁肽联用二甲双胍在降低TNF-α、VEGF和P38MAPK mRNA表达方面效果更优。结论:(1)利拉鲁肽与二甲双胍联合用药作用于高糖高脂+STZ诱导的糖尿病模型大鼠,能显著改善血糖、脂质代谢紊乱、胰岛素抵抗等情况,能增加糖尿病大鼠的体质量,且效果优于二甲双胍单用药治疗。(2)利拉鲁肽与二甲双胍联合用药作用于高糖高脂+STZ诱导的糖尿病模型大鼠,可降低血清中TNF-α、VEGF、VWF含量,且效果优于单用药治疗。(3)利拉鲁肽与二甲双胍联合用药作用于高糖高脂+STZ诱导的糖尿病模型大鼠,可降低胸主动脉胶原容积分数、改善血管内皮损伤,降低血管纤维化水平,其机制可能与下调TNF-α、VEGF、P38MAPK表达有关。
[Abstract]:Objective: To observe the effects of the combination of the combination of Li Lu Lu peptide and metformin on blood glucose, blood lipid and insulin resistance in diabetic rats and the effect on the thoracic aorta fibrosis and its possible mechanism. Methods: 9 SPF grade SD male rats were randomly selected as normal control group (group NG, double steam water, 5 ml. Kg-1. D-1, i.g.), and the other rats were fed high fat and high fat diet. The diabetic rat model was induced by intraperitoneal injection of streptozotocin (STZ) 35mg / kg-1. The rats were randomly divided into the diabetic model control group (DM group, double steam water, 5 ml kg-1. D-1, i.g.), the treatment group of metformin (Met group, 100 mg, kg-1.), and the treatment group (0.2). D-1, i.p.), the treatment group (group Lir+Met, 0.1 mg, kg-1, D-1, i.p.+50 mg, kg-1 D-1, i.g.), glitazide combined with metformin treatment group (Gli+Met group, 10) at 8 o'clock every morning or intraperitoneal injection for 6 weeks. Every 2 weeks. The rats were weighed once, and the rats' fasting blood glucose (FBG) changes were measured. The oral glucose tolerance (OGTT) test was conducted at the end of the 6 week. The area under the curve (AUC) was calculated. The abdominal aorta was taken from the abdominal aorta, the blood lipid level, the glycated hemoglobin (HbA1c) and the fasting serum insulin (FINS) level were measured, and the insulin resistance index (IR) was calculated, and the cause of the bad cause of the tumor in the serum was measured. TNF- alpha, vascular endothelial growth factor (VEGF) and vascular hemophilia factor (VWF) in rats; HE staining was used to observe the pathological injury of thoracic aorta in rats; Masson staining method was used to observe the changes of collagen fiber in thoracic aorta and measure collagen volume (CVF); RT-PCR method was used to detect TNF- alpha, VEGF, mitogen activated protein kinase p38 (P38MAPK) mRNA expression. (1) compared with the normal group, the body weight (BW) and high density lipoprotein cholesterol (HDL-C) in the diabetic model group were significantly decreased (P0.01), while FBG, FINS, TG, TC, LDL-C, HbA1c increased significantly (P0.01 or P0.05), suggesting the emergence of significant insulin resistance. Thickening, endothelial cells, smooth muscle arrangement disorder, inflammatory infiltration, obvious pathological damage, Masson staining of collagen fibrous deposition, vascular thickening, CVF significantly increased (P0.01); RT-PCR assay showed that the expression of TNF- alpha, VEGF and P38MAPK mRNA increased (P0.01 or P0.05). (2) BW, H group rats BW, H DL-C increased (P0.01 or P0.05), FBG, FINS, TG, LDL-C, HbA1c had a different degree of decline (P0.01 or P0.05), HOMA-IR decreased (P0.01 or decreased), and the effect of alalu peptide combined with metformin was better than metformin single drug group. The effect of guanidine group was significant (P0.01 or P0.05); HE staining showed that the aorta arteritis of the rats in each group was improved, the hyperplasia of endometrium was improved, the pathological damage was reduced, and the effect of the drug group was better than that of the single drug group, and the Masson staining showed that the group could reduce the deposition of collagen fiber in the thoracic aorta and the blood vessels. The effect of thickening and other phenomena significantly decreased CVF (P0.01). The effect of alalup combined with metformin on the reduction of CVF was equivalent to that of the control group. The RT-PCR method showed that the expression of TNF- alpha, VEGF and P38MAPK mRNA decreased (P0.01 or P0.05) in each drug group, while the use of two metformin was better in reducing TNF- alpha. Conclusion: (1) the combination of L / M and metformin on diabetic rats induced by high glucose and high fat +STZ can significantly improve the blood glucose, lipid metabolism disorder, insulin resistance and so on. It can increase the body mass of diabetic rats, and the effect is better than metformin alone. (2) the effect of the combination of the use of Li Lu and metformin in combination with metformin. The diabetic rat model induced by high glucose and high fat +STZ can reduce the content of TNF- alpha, VEGF, and VWF in serum, and the effect is better than that of single drug treatment. (3) the combination of alalu and metformin on high glucose and high fat +STZ induced diabetic rats can reduce the volume fraction of collagen in the thoracic aorta, improve vascular endothelial damage and reduce vascular fiber. Its mechanism may be related to downregulation of TNF-, VEGF and P38MAPK expression.

【学位授予单位】：皖南医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R587.1

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