利拉鲁肽通过调节microRNA-375对胰岛细胞凋亡的影响

发布时间：2018-08-20 16:43

【摘要】：目的:观察利拉鲁肽通过微小RNA-375(microRNA-375,miR-375)对db/db小鼠胰岛细胞凋亡的影响,探讨其可能作用机制,为临床应用提供药效学依据。方法:20只8周龄雄性db/m小鼠为正常对照组,皮下注射等量的生理盐水。40只8周龄雄性db/db小鼠随机分为2组,每组20只:糖尿病对照组的db/db小鼠皮下注射等量生理盐水;利拉鲁肽组的db/db小鼠皮下注射利拉鲁肽300μg·kg~(-1)·d~(-1)。给药8周后,检测各组小鼠体重(BW)、空腹血糖(FBG)、空腹胰岛素(FINS)、甘油三酯(TG)、总胆固醇(TC)及低密度脂蛋白胆固醇(LDL-C)含量,并进行腹腔注射葡萄糖耐量实验(IPGTT)和胰岛素耐量实验(ITT);苏木精-伊红(HE)染色检测胰岛组织病理学变化;原位末端转移酶标记技术(TUNEL)检测胰岛凋亡情况;Western blot法检测胰岛凋亡相关蛋白caspase-3、Bcl-2及Bax的蛋白水平;实时荧光定量PCR检测胰岛miR-375的表达水平。小鼠胰岛β细胞系MIN-6分为对照组(等量溶媒孵育)、miRNA-375 mimic组和miRNA-375 mimic+利拉鲁肽组,MTT实验检测细胞活力,Western blot法检测各组细胞caspase-3、Bcl-2及Bax的蛋白水平。结果:整体实验中,与对照组相比,利拉鲁肽组BW、FBG、FINS、TC、TG及LDL-C含量明显降低(P0.05);胰岛数量较模型组增多,体积较大,细胞结构明显改善;胰岛细胞凋亡减少;利拉鲁肽组的Bcl-2表达明显增高,caspase-3和Bax的表达显著下降(P0.05);胰岛组织miR-375的水平显著降低(P0.01)。细胞实验中,经miRNA-375 mimic处理24 h后,MIN-6细胞活力显著降低,Bcl-2表达减少,而caspase-3和Bax的蛋白水平显著增加(P0.05),给予利拉鲁肽组治疗后,MIN-6细胞活力明显上升,Bcl-2表达明显增高,caspase-3和Bax的蛋白水平显著下降(P0.05)。结论:利拉鲁肽可以抑制糖尿病胰岛β细胞的凋亡,其机制可能与调控胰岛组织中miR-375的表达有关。
[Abstract]:Aim: to observe the effect of liralutin on the apoptosis of islet cells in db/db mice by microRNA-375 (microRNA-375 miR-375), to explore the possible mechanism of its action, and to provide pharmacodynamic basis for clinical application. Methods Twenty 8-week-old male db/m mice were randomly divided into two groups: normal control group (n = 20) and normal control group (n = 20). 40 8-week-old male db/db mice were subcutaneously injected with the same amount of normal saline. Db/db mice were subcutaneously injected with Lilaru peptide 300 渭 g kg ~ (-1) d ~ (-1). After 8 weeks of administration, fasting blood glucose (BW), fasting blood glucose (FBG), fasting insulin, (FINS), triglyceride, (TG), total cholesterol, (TC) and low density lipoprotein cholesterol (LDL-C) were measured in each group. The histopathological changes of pancreatic islets were detected by intraperitoneal glucose tolerance test (IPGTT) and insulin tolerance test (ITT);) with hematoxylin and eosin (HE) staining. In situ terminal transferase labeling (TUNEL) was used to detect the apoptosis of pancreatic islets. Western blot was used to detect the protein levels of caspase-3, Bcl-2 and Bax, and real-time quantitative PCR was used to detect the expression of miR-375. Murine islet 尾 cell line MIN-6 was divided into two groups: control group (equal solvent incubated) and miRNA-375 mimic group. The cell viability was detected by Western blot assay. The protein levels of caspase-3, Bcl-2 and Bax were detected by Western blot. Results: in the whole experiment, compared with the control group, the content of TG and LDL-C in BWFBGG FINSN TCU group was significantly lower than that in the control group (P0.05), the number of islets increased, the volume of islets was larger, the cell structure was improved obviously, and the apoptosis of islet cells was decreased. The expression of caspase-3 and Bax in Lilaru group was significantly higher than that in control group (P0.05), and the level of miR-375 in islet tissue was significantly decreased (P0.01). In the cell experiment, the activity of MIN-6 cells decreased significantly after 24 h of miRNA-375 mimic treatment, and the expression of Bcl-2 decreased significantly. However, the protein levels of caspase-3 and Bax increased significantly (P0.05), and the activity of MIN-6 cells increased significantly (P0.05). The expression of Bcl-2 increased significantly (P0.05). Conclusion: Lilaru peptide can inhibit the apoptosis of diabetic islet 尾 cells, and its mechanism may be related to the regulation of miR-375 expression in islet tissues.
【作者单位】：郑州人民医院药学部;郑州大学附属肿瘤医院河南省肿瘤医院药学部;
【分类号】：R587.1

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