miR-221对糖尿病肾病小鼠胰岛β细胞功能的保护作用及其机制

doi:10.13481/j.1671-587x.20160513

吉林大学学报(医学版) ›› 2016, Vol. 42 ›› Issue (05): 905-909.doi: 10.13481/j.1671-587x.20160513

miR-221对糖尿病肾病小鼠胰岛β细胞功能的保护作用及其机制

李俊, 代留玲, 李晔, 茶春丽

昆明医科大学第一附属医院肾内科, 云南昆明 650032

收稿日期:2015-09-17 出版日期:2016-09-28 发布日期:2016-09-29
通讯作者: 李俊,主任医师,硕士研究生导师(Tel:0871-65324888-2528,E-mail:dllijun1968@163.com) E-mail:dllijun1968@163.com
作者简介:李俊(1968-),女,白族,云南省大理市人,主任医师,医学硕士,主要从事肾纤维化临床与基础方面的研究。
基金资助:
云南省科技厅应用基础研究计划项目资助课题（2012FB029）；云南省教育厅资助项目资助课题（2014J040）

Protective effect of miR-221 on β cells in mice with diabetic nephropathy and its mechanism

LIJun, DAILiuling, LI Ye, CHA Chunli

Department of Internal Medicine, First Affiliated Hospital, Kunming Medical University, Kunming 650032, China

Received:2015-09-17 Online:2016-09-28 Published:2016-09-29

摘要/Abstract

摘要：

目的：研究miR-221对糖尿病肾病（DN）小鼠胰岛β细胞功能的影响，阐明miR-221在DN中的保护作用机制。方法：8周龄野生型雄性C57BL/6J小鼠20只随机分成对照组和DN组，每组10只，对照组小鼠给予正常饮食，DN组小鼠给予高脂饮食，并注射100 mg·kg^-1链脲佐菌素（STZ）诱导部分胰岛素缺陷，对照组小鼠注射同体积柠檬酸缓冲液。之后DN组继续给予高脂饮食，对照组给予正常饮食，饲养10周后收集小鼠血液和尿液，检测血糖、血肌酐、血尿素氮浓度和24 h尿白蛋白排泄率等生理参数。在DN组小鼠中分离得到胰岛细胞。利用Real-time PCR检测胰岛细胞中SOCS3 mRNA表达水平，采用MTT法检测胰岛细胞增殖情况，ELISA法检测胰岛细胞中胰岛素含量和胰岛素的释放水平，荧光素酶报告基因法检测SOCS3活性荧光酶素报告基因活性。结果：成功构建DN小鼠模型。DN组小鼠的血糖、血肌酐、血尿素氮浓度和24h尿白蛋白排泄率均高于对照组（P<0.05）。过表达miR-221后，DN组小鼠胰岛细胞中SOCS3 mRNA表达水平低于正常胰岛细胞（P<0.05），过表达miR-221后胰岛细胞的增殖能力低于正常胰岛细胞（P<0.05），荧光素酶报告基因法确定SOCS3为miR-221下游靶基因。过表达miR-221后，胰岛细胞中胰岛素质量分数和胰岛素释放水平均高于正常胰岛细胞（P<0.05）。结论：miR-221通过下调SOCS3水平促进胰岛细胞合成和分泌胰岛素的功能，改善DN小鼠中胰岛细胞的功能障碍。

关键词: 糖尿病肾病, miR-221, 细胞因子信号抑制因子 3, 胰岛&beta, 细胞

Abstract:

Objective: To study the influence of miR-221 in the β cells of mice with diabetic nephropathy (DN), and to clarify the protective effect of miR-221 on DN. Methods: Twenty wild type C57BL/6J mice aged 8 weeks were divided into control group and DN group(n=10).The DN mice models were constructed with 14 weeks of high fat diet,and 100mg·kg^-1 streptozotocin (STZ) was used to induce the partial insulin deficiency. 10 weeks later the blood glucose, serum creatinine, blood urea nitrogen content and 24h-urinary albumin excretion rate were detected. The β cells of islet were isolated from the DN mice. The expression level of SOCS3 mRNA in β cells of islet was valued by Real-time PCR. The proliferation of pancreatic β cells of islet was examined by MTT assay. The contents of insulin and insulin release levels in pancreatic β cells of islet were detected by ELISA assay. Luciferase reporter gene assay was used to detect the luciferase reporter gene activity of SOCS3. Results: The DN mouse models were constructed successfully. The blood glucose, serum creatinine, blood urea nitrogen and 24h-urinary albumin excretion rate of the mice in DNA group were higher than those in control group(P<0.05). After overexpression of miR-221, the expression level of SOCS3 mRNA in pancreatic β cells of islet and the proliferation ability of β cells of islet of the mice in DN group were lower than those of normal islet cells (P<0.05). Then luciferase reporter gene method found SOCS3 as one of the target genes of miR-221. After overexpression of miR-221, the insulin release level and insulin content in pancreatic β cells of islet were higher than those of normal islet cells(P<0.05). Conclusion: miR-221 can promopt the synthesis and secretion of β cells of islet and inhibit the dysfunction of β cells of islet in the DN mice by down-regulating the SOCS3 level.

Key words: diabetic nephropathy, miR-221, suppressor of cytokine signaling 3, &beta, cells of islet

中图分类号:

R587.1

李俊, 代留玲, 李晔, 茶春丽. miR-221对糖尿病肾病小鼠胰岛β细胞功能的保护作用及其机制[J]. 吉林大学学报(医学版), 2016, 42(05): 905-909.

LIJun, DAILiuling, LI Ye, CHA Chunli. Protective effect of miR-221 on β cells in mice with diabetic nephropathy and its mechanism[J]. Journal of Jilin University Medicine Edition, 2016, 42(05): 905-909.

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miR-221对糖尿病肾病小鼠胰岛β细胞功能的保护作用及其机制

Protective effect of miR-221 on β cells in mice with diabetic nephropathy and its mechanism

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