利拉鲁肽通过microRNA-33对2型糖尿病小鼠肝损伤的治疗作用

发布时间：2018-05-01 03:01

本文选题：利拉鲁肽 + 微小RNA-　；参考：《中国病理生理杂志》2017年01期

【摘要】：目的:观察利拉鲁肽对2型糖尿病小鼠肝组织微小RNA-33(microRNA-33,miR-33)、腺苷酸活化蛋白激酶(AMPK)及肝细胞凋亡通路相关蛋白的影响,探讨其可能作用机制,为临床应用提供药效学依据。方法:采用高脂饮食联合链脲佐菌素(STZ)腹腔注射复制C57BL/6小鼠2型糖尿病模型。小鼠随机分为4组,每组15只:对照组为正常小鼠皮下注射等量的生理盐水;模型组为T2DM小鼠皮下注射等量的生理盐水;利拉鲁肽低剂量组为了2DM小鼠皮下注射100μg·kg~(-1)·d~(-1)利拉鲁肽;利拉鲁肽高剂量组为T2DM小鼠皮下注射200μg·kg~(-1)·d~(-1)利拉鲁肽。给药4周后进行口服葡萄糖耐量试验(OGTT),检测各组小鼠空腹血糖(FBG)、甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、丙氨酸氨基转移酶(ALT)及天冬氨酸氨基转移酶(AST)的含量;HE染色检测肝组织病理学变化;免疫荧光检测肝组织的cleaved caspase-3;Western blot法检测p-AMPK/AMPK及凋亡相关蛋白Bcl-2、caspase-3的水平;实时定量PCR检测肝组织miR-33的水平。结果:与模型组相比,利拉鲁肽高、低剂量组小鼠FBG、TG、TC、LDL-C、ALT及AST含量明显降低,HDL-C含量明显升高(P0.05);肝组织病理结构的紊乱得到明显改善;免疫荧光结果可见,利拉鲁肽高、低剂量给药组小鼠的cleaved caspase-3明显降低;Western blot实验结果可见,利拉鲁肽高、低剂量给药组小鼠肝组织的Bcl-2表达明显增高,caspase-3表达显著下降(P0.05)肝组织AMPK磷酸化水平显著增加(P0.01);且肝组织miR-33水平显著降低(P0.01),且呈一定的剂量依赖性。结论:利拉鲁肽可以缓解2型糖尿病小鼠肝损伤,其机制可能与降低肝组织miR-33表达,从而增加AMPK磷酸化水平,进而抑制肝细胞凋亡有关。
[Abstract]:Aim: to observe the effects of liverudin on minimal RNA-33 miR-33 microRNA-33 miR-33, adenylate activated protein kinase (AMPKK) and apoptosis-related proteins related to hepatocyte apoptosis in type 2 diabetic mice, and to explore its possible mechanism, and to provide pharmacodynamics basis for clinical application. Methods: type 2 diabetic model of C57BL/6 mice was induced by high fat diet combined with streptozotocin (STZ) intraperitoneal injection. Mice were randomly divided into 4 groups: normal mice were subcutaneously injected with the same amount of normal saline in the control group, T2DM mice were subcutaneously injected with the same amount of normal saline in the model group, and the 2DM mice were subcutaneously injected with 100 渭 g / kg of Lilarudine in the low dose group. In the high dose group, T2DM mice were subcutaneously injected with 200 渭 g / kg of Lilaru peptide. After 4 weeks of administration, oral glucose tolerance test (OGTT) was used to detect fasting blood glucose (FBGG), triglyceride (TGG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), alanine aminotransferase (alt) and asparagus. The content of aminoaminotransferase (AST) and HE staining were used to detect the histopathological changes of liver. The levels of p-AMPK/AMPK and Bcl-2 caspase-3 were detected by cleaved caspase-3 Western blot and real-time quantitative PCR was used to detect the level of miR-33 in liver tissue. Results: compared with the model group, the levels of alt and AST in the low dose group were significantly lower than those in the model group, the contents of HDL-C and AST were significantly increased, the pathological structure of liver was obviously improved, and the results of immunofluorescence showed that Lilaru peptide was high. The results of low dose administration group showed that the cleaved caspase-3 of mice was significantly lower than that of control group. The results of Western blot test showed that Lilaru peptide was higher than that in control group. In low dose group, the expression of Bcl-2 in liver tissue increased significantly (P 0.05) the level of AMPK phosphorylation in liver tissue increased significantly, and the level of miR-33 in liver tissue decreased significantly in a dose-dependent manner. Conclusion: Lilaru peptide can alleviate liver injury in type 2 diabetic mice, and its mechanism may be related to the decrease of miR-33 expression in liver tissue, thereby increasing the level of AMPK phosphorylation and inhibiting hepatocyte apoptosis.
【作者单位】：郑州大学附属肿瘤医院河南省肿瘤医院药学部;郑州人民医院药学部;
【分类号】：R587.2;R575

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