[1]林晓璐,张红,夏小慧,等.Exendin-4对高糖培养肾小球系膜细胞 ECM分泌的影响及相关机制[J].国际内分泌代谢杂志,2016,36(05):302-307.[doi:10.3760/cma.j.issn.1673-4157.2016.05.05]
　Lin Xiaolu*,Zhang Hong,Xia Xiaohui,et al.Effects and mechanism of exendin-4 on extracellular matrix secretion of mesangial cells cultured in high glucose[J].International Journal of Endocrinology and Metabolism,2016,36(05):302-307.[doi:10.3760/cma.j.issn.1673-4157.2016.05.05]
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Exendin-4对高糖培养肾小球系膜细胞 ECM分泌的影响及相关机制()
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《国际内分泌代谢杂志》[ISSN:1673-4157/CN:12-1383/R]

卷:
36
期数:
2016年05期
页码:
302-307
栏目:
论著
出版日期:
2016-09-20

文章信息/Info

Title:
Effects and mechanism of exendin-4 on extracellular matrix secretion of mesangial cells cultured in high glucose
作者:
林晓璐; 张红; 夏小慧; 陈娟; 张日东; 章向成; 李伟
221000 徐州医学院研究生院(林晓璐,夏小慧); 223001 南京医科大学附属淮安第一医院内分泌科(陈娟,张日东,章向成,张红); 221000 徐州医学院附属医院内分泌科(李伟)
Author(s):
Lin Xiaolu*;  Zhang Hong;  Xia Xiaohui;  Chen Juan;  Zhang Ridong;  Zhang Xiangcheng;  Li Wei
*Graduated School of Xuzhou Medical College,Xuzhou 221000,China
关键词:
Exendin-4;  肾小球系膜细胞;  炎症;  核因子-κB
Keywords:
Exendin-4;  Glomerular mesangial cells;  Inflammation;  Nuclear factor-κB
DOI:
10.3760/cma.j.issn.1673-4157.2016.05.05
摘要:
目的 研究exendin-4对高糖培养大鼠肾小球系膜细胞(GMCs)细胞外基质分泌的影响及相关机制。方法 体外培养GMCs,分为4组:正常对照组(NC组)、正常对照+exendin-4组(NCE组)、高糖组(HG组)、高糖+exendin-4组(HGE组); HGE组细胞分别给予3,5,10,15,30 nmol/L exendin-4培养12,24,48 h,采用细胞增殖与活性检测试剂盒(Cell Counting Kit-8,CCK8)法测定细胞增殖情况,确定exendin-4的最适作用时间与浓度。ELISA测定细胞上清中细胞外基质蛋白纤维连接蛋白(FN)和Ⅳ型胶原蛋白水平。RT-PCR检测FN、Ⅳ型胶原蛋白及单核细胞趋化蛋白-1(MCP-1)、肿瘤坏死因子-α(TNF-α)、细胞间黏附分子-1(ICAM-1)、转化生长因子-β1(TGF-β1)mRNA的表达。Western印迹测定核因子-κB的表达。结果(1)高糖培养的GMCs给予10 noml/L exendin-4培养24 h后,其细胞增殖率较3 nmol/L、5 nmol/L的exendin-4明显降低(F=120.808, P<0.05); 与15 nmol/L、30 nmol/L相比,10 noml/L(HGE组)的exendin-4对GMCs增殖率无明显变化(P均>0.05)。(2)与NC组、NCE组相比,HG组FN、Ⅳ型胶原蛋白分泌及mRNA以及MCP-1、TNF-α、ICAM-1、TGF-β1 mRNA的表达均显著升高(F=6.894～166.914, P均<0.05); 与HG组相比,HGE组FN、Ⅳ型胶原蛋白分泌及mRNA以及TNF-α、MCP-1、ICAM-1、TGF-β1 mRNA表达量明显降低(F=6.894～166.914,P均<0.05)。(3)与NC组、NCE相比,HG组核因子-κB蛋白的表达明显增多(F=133.1,P<0.05)。与HG组相比,HGE组核因子-κB蛋白的表达则明显受到抑制(F=133.1,P<0.05)。结论 Exendin-4抑制高糖培养GMCs的细胞外基质分泌,与其抑制核因子-κB介导的炎性反应有关。
Abstract:
Objective To explore the effects and mechanism of exendin-4 on the secretion of extracellular matrix in high glucose-cultured glomerular mesangial cells(GMCs). Methods GMCs were cultured in vitro and divided into 4 groups: normal control(NC)group, normal control with exendin-4(NCE)group, high glucose(HG)group, high glucose with exendin-4(HGE)group. Cells in HGE group were treated with 3, 5, 10, 15 or 30 nmol/L exendin-4 for about 12, 24 or 48 hours, respectively. The proliferation and cell activity of GMCs were used to assess the most suitable culture time and concentration using Cell Counting Kit-8. The levels of fibronectin(FN)and collagen type Ⅳ in the cell supernatant were measured by ELISA. The levels of collagen type Ⅳ, FN, monocyte chemotactic protein-1(MCP-1), tumor necrosis factor(TNF)-α, intercellular cell adhesion molecule-1(ICAM-1), transforming growth factor-β1(TGF-β1)were evaluated by RT-PCR. The expression of nuclear factor-κB(NF-κB)in each group was analyzed by Western blotting. Results(1)After treated by 10 nmol/L exendin-4 for 24 hours, the growth rate of GMCs cultured in high glucose was significantly decreased compared with 3 nmol/L or 5 nmol/L(F=120.808, P<0.05), but not different from 15 nmol/L or 20 nmol/L(all P>0.05).(2)Compared with NC group and NCE group,the expression of FN, collagen type Ⅳ, as well as their mRNA level and the inflammatory mediators such as MCP-1, TNF-α, ICAM-1 and TGF-β1 were significantly increased in HG group(F=6.894-166.914, all P<0.05). Compared with HG group, the protein and mRNA level of FN and collagen type Ⅳ, and the expression of TNF-α, MCP-1, ICAM-1 and TGF-β1 were inhibited greatly in HGE group(F=6.894-166.914, all P<0.05).(3)Compared with NC group and NCE group, the expression of NF-κB increased significantly in HG group(F=133.1,P<0.05). Compared with HG group, the expression of NF-κB was suppressed significantly in HGE group(F=133.1, P<0.05). Conclusion Exendin-4 inhibits the secretion of extracellular matrix of GMCs cultured in high glucose, and the mechanism is associated with the inhibition of inflammation induced by NF-κB.

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备注/Memo

备注/Memo:
基金项目:国家自然科学基金资助项目(81200595,81400807); 江苏省卫计委科研项目(H201253) 通信作者:李伟,Email:Liwei.190@hotmail.com
更新日期/Last Update: 2016-11-20