抑制miR-200c表达对糖尿病肾病大鼠肾组织的保护作用及机制研究

doi:10.13418/j.issn.1001-165x.2019.06.012

中国临床解剖学杂志 ›› 2019, Vol. 37 ›› Issue (6): 668-672.doi: 10.13418/j.issn.1001-165x.2019.06.012

抑制miR-200c表达对糖尿病肾病大鼠肾组织的保护作用及机制研究

刘新磊¹，　宋卫敏¹，　高志强¹，　陈福莲²

潍坊医学院附属益都中心医院　1.肾内科，2.内分泌科，山东潍坊 262500

收稿日期:2019-03-11 出版日期:2019-11-25 发布日期:2019-12-02
通讯作者: 陈福莲，E-mail：chenfulian2000@yeah.net
作者简介:刘新磊（1982-），本科，护师，研究方向：糖尿病肾病预防，E-mail：liuxinleiwf@yeah.net
基金资助:
山东省医药卫生科技发展计划面上项目（2017WSA07016）

Protection effects and the mechanism on renal tissue in rats with diabetie nephropathy through inhibiting miR-200c expression

LIU Xin-lei¹, SONG Wei-min¹, GAO Zhi-qiang¹, CHEN Fu-lian²

1. Department of Nephrology, 2. Department of Endocrine, Yi Du Central Hospital Affiliated to Weifang Medical University, Weifang 262500, Shandong Province, China

Received:2019-03-11 Online:2019-11-25 Published:2019-12-02

摘要/Abstract

摘要： 目的　探讨抑制miR-200c表达对糖尿病肾病（diabetic nephropathy，DN）SD大鼠肾的保护作用及机制。方法　采用高糖高脂饮食联合链脲佐菌素（streptozotocin，STZ）腹腔注射诱导建立SD大鼠DN模型，将造模成功的30只DN大鼠随机分为模型组和观察组，每组15只，同时取15只正常健康的SD大鼠作为对照组，造模成功后每7 d给予观察组大鼠尾静脉注射antagomir-200c（30 mg/kg），模型组和对照组给予尾静脉注射等量的生理盐水。8周后，检测大鼠血清肌酐（Cr）、尿素氮（BUN）和24 h尿蛋白定量水平，实时荧光定量PCR（qRT-PCR）检测肾组织中miR-200c的表达，HE染色观察肾组织病理学变化，活性氧簇（ROS）和丙二醛（MDA）试剂盒检测肾组织中ROS和MDA水平，Western blot检测肾组织中转化生长因子-β1（TGF-β1）、纤连蛋白（fibronectin）的水平。结果　与对照组比较，模型组大鼠肾组织miR-200c表达、血清中BUN、Cr水平、24 h尿蛋白定量、肾间质损伤评分、ROS、MDA水平及TGF-β1、fibronectin蛋白表达均升高（P<0.05）。与模型组比较，观察组大鼠以上指标均降低（P<0.05）。结论　miR-200c在STZ诱导的DN大鼠肾组织中表达升高，抑制miR-200c能够对DN大鼠的肾起到一定保护作用，可能与降低肾组织的氧化应激水平和对TGF-β1信号通路的抑制有关。

关键词: 大鼠, 　糖尿病肾病, 　miR-200c, 　氧化应激, 　TGF-β1

Abstract: Objective　To explore the protection on the kidneys of streptozotocin (STZ)-induced diabetic nephropathy (DN) SD rats, and the related mechanism, by abating miR-200C.　Methods　The DN SD rat model was constructed through high-glucose and high-fat diet combined with intraperitoneal injection of STZ. Thirty successfully constructed DN model rats were randomly divided into a model group and an observation group, with 15 rats in each group. Additionally, 15 normal healthy SD rats were collected as the control group. After successful modeling, rats in observation group were given a tail vein injection of antagomir-200c (30 mg/kg) every 7 days, while those in model group and control group were given a tail vein injection of equivalent amounts of normal saline. Eight weeks later, the serum creatinine (Cr), blood urea nitrogen (BUN), and 24 h urine protein quantitation (UPQ) level in rats were detected. Meanwhile, miR-200c expression in renal tissues was detected through real-time fluorescence quantitative PCR (qRT-PCR). Histopathological changes in kidneys were observed through HE staining. Reactive oxygen species (ROS) and malondialdehyde (MDA) levels in renal tissues were detected using the ROS and MDA detection kits. In addition, the protein levels of transforming growth factor-β1 (TGF-β1) and fibronectin in renal tissues were also detected through Western blot. Results Compared with the control group, miR-200c expression in renal tissues, serum BUN and Cr levels, 24 h UPQ, renal interstitial injury score, ROS and MDA levels, as well as TGF-β1 and fibronectin protein expression in the model group were all markedly elevated (P＜0.05). Compared with the model group, the above indicators in the observation group were all markedly reduced (P＜0.05). Conclusions miR-200c expression in the renal tissues of STZ-induced DN rats is up-regulated, and abation of miR-200c can partly protect the kidneys of DN rats, which may be related to the reduction of oxidative stress in kidney tissue and the inhibition of TGF-β1 signaling pathway.

Key words: Rat; , , Diabetic nephropathy; , , miR-200c; , , Oxidative stress; , , TGF-β1

中图分类号:

R587.2

刘新磊, 宋卫敏, 高志强, 陈福莲. 抑制miR-200c表达对糖尿病肾病大鼠肾组织的保护作用及机制研究[J]. 中国临床解剖学杂志, 2019, 37(6): 668-672.

LIU Xin-lei, SONG Wei-min, GAO Zhi-qiang, CHEN Fu-lian. Protection effects and the mechanism on renal tissue in rats with diabetie nephropathy through inhibiting miR-200c expression[J]. Chinese Journal of Clinical Anatomy, 2019, 37(6): 668-672.

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抑制miR-200c表达对糖尿病肾病大鼠肾组织的保护作用及机制研究

Protection effects and the mechanism on renal tissue in rats with diabetie nephropathy through inhibiting miR-200c expression

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