二甲双胍抑制高糖培养大鼠肾小球系膜细胞内P38MAPK信号通路与氧化应激

doi:10.12092/j.issn.1009-2501.2018.06.003

中国临床药理学与治疗学 ›› 2018, Vol. 23 ›› Issue (6): 614-620.doi: 10.12092/j.issn.1009-2501.2018.06.003

二甲双胍抑制高糖培养大鼠肾小球系膜细胞内P38MAPK信号通路与氧化应激

姚新明¹，孔祥¹，赵咏莉¹，华强¹，何春玲¹，余丹²，李业琼²，戴晓康²，叶山东²

¹皖南医学院弋矶山医院内分泌科，芜湖 241001，安徽；²安徽省立医院内分泌科，合肥 230001，安徽

收稿日期:2018-01-22 修回日期:2018-04-11 出版日期:2018-06-26 发布日期:2018-06-19
通讯作者: 叶山东，男，博士，教授，博士生导师，主要研究方向：糖尿病肾脏疾病。 E-mail: ysd196406@163.com
作者简介:姚新明，男，博士，主要研究方向：糖尿病肾脏疾病。 E-mail: yxm6965@sina.com
基金资助:
安徽省自然科学基金(1608085QH191) ；安徽省高校优秀青年人才支持计划项目（gxyq2017035）；皖南医学院弋矶山医院人才引进基金（YR201507）；安徽省自然科学基金（1508085SMH227）

Metformin inhibits p38MAPK signaling pathway and oxidativestress in rat glomerular mesangial cells cultured with high glucose

YAO Xinming¹ , KONG Xiang¹, ZHAO Yongli ¹, HUA Qiang ¹, HE Chunling¹, YU Dan², LI Yeqiong², DAI Xiaokang², YE Shandong²

¹Department of Endocrinology, Yijishan Hospital Affiliated to Wannan Medical College, Wuhu 241001, Anhui, China；² Department of Endocrinology, Anhui Provincial Hospital, Hefei 230001, Anhui, China

Received:2018-01-22 Revised:2018-04-11 Online:2018-06-26 Published:2018-06-19

摘要/Abstract

摘要：

目的：观察高浓度葡萄糖培养大鼠肾小球系膜细胞内p38丝裂原活化蛋白激酶(p38 mitogenactivated protein kinase，p38MAPK)信号通路与氧化应激的关系及二甲双胍的调控作用。方法：大鼠肾小球系膜细胞(HBZY1)在完全培养基中传代培养，并分成以下5组:（1）正常对照组(NC组)；（2）高浓度葡萄糖培养组(HG组)；（3）二甲双胍治疗组(MET组)；（4）SB203580治疗组(SB组)；（5）N-乙酰半胱氨酸治疗组(NAC)组。流式细胞术检测大鼠肾小球系膜细胞内活性氧(ROS)水平。用比色法和ELISA法分别检测细胞上清液中超氧化物歧化酶(SOD)活性与丙二醛(MDA)含量。实时定量PCR法检测肾小球系膜细胞内P22phox mRNA表达。Western blot法检测肾小球系膜细胞内P22phox蛋白和磷酸化p38MAPK(p-p38MAPK)蛋白表达。结果:与正常对照组相比，高浓度葡萄糖培养组细胞上清液中SOD活性降低，相反细胞上清液中MDA含量、肾小球系膜细胞内ROS水平、P22phox mRNA和蛋白及磷酸化p38MAPK蛋白表达均上升(P<0.05)。当二甲双胍干预后，细胞上清液中SOD活性上升,同时细胞上清液中MDA含量、肾小球系膜细胞内ROS水平、P22phox mRNA和蛋白及磷酸化p38MAPK蛋白表达均下降(P<0.05)。相似的结果出现在SB203580或N-乙酰半胱氨酸干预后(P<0.05)。结论:二甲双胍可以减轻高浓度葡萄糖培养大鼠肾小球系膜细胞氧化应激和磷酸化p38MAPK蛋白表达，进而起到了保护肾脏的作用。

关键词: 二甲双胍, 氧化应激, p38MAPK, 肾小球系膜细胞

Abstract:

AIM: To observe relationship between p38MAPK signaling pathway and oxidative stress in rat glomerular mesangial cells (MCs) cultured in high glucose medium and the regulation of metformin. METHODS: Rat glomerular MCs were cultured in a complete medium, and were divided into five groups: normal control group (group NC), high glucose group(group HG) , metformintreated group (group MET), SB203580treated group (group SB), N-acetylcysteinetreated group (group NAC). Intracellular reactive oxygen species (ROS) content of rat glomerular MCs was tested by flow cytometry. Superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in supernatant fluid were detected by colorimetry and ELISA, respectively. P22phox mRNA of rat glomerular MCs was determined by real-time quantitative PCR. P22phox protein and phosphorylation of p38MAPK(p-p38MAPK)protein of rat glomerular MCs were detected by Western blot. RESULTS: Compared with group NC, the activity of SOD was decreased, whereas the level of MDA, intracellular ROS content, P22phox mRNA and protein, p-p38MAPK protein expression were increased in group HG (P<0.05). When metformin was added to the high glucose medium, the activity of SOD was increased, meanwhile,the level of MDA, intracellular ROS content, P22phox mRNA and protein,pp38MAPK protein expression were decreased (P<0.05), and the results were similar with SB203580 or N-acetylcysteine was added to the high glucose medium (P<0.05). CONCLUSION: Metformin can alleviate high glucose induced oxidative stress and p-p38MAPK protein expression of rat glomerular MCs,which maybe contribute to its renoprotection.

Key words: metformin, oxidative stress, p38 MAPK, mesangial cells

中图分类号:

R965.2

姚新明，孔祥，赵咏莉，华强，何春玲，余丹，李业琼，戴晓康，叶山东. 二甲双胍抑制高糖培养大鼠肾小球系膜细胞内P38MAPK信号通路与氧化应激[J]. 中国临床药理学与治疗学, 2018, 23(6): 614-620.

YAO Xinming, KONG Xiang, ZHAO Yongli, HUA Qiang, HE Chunling, YU Dan, LI Yeqiong, DAI Xiaokang, YE Shandong. Metformin inhibits p38MAPK signaling pathway and oxidativestress in rat glomerular mesangial cells cultured with high glucose[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2018, 23(6): 614-620.

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二甲双胍抑制高糖培养大鼠肾小球系膜细胞内P38MAPK信号通路与氧化应激

Metformin inhibits p38MAPK signaling pathway and oxidativestress in rat glomerular mesangial cells cultured with high glucose

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