上海交通大学学报（医学版）

›› 2010, Vol. 30 ›› Issue (5): 518-.

• 论著（基础研究） • 上一篇    下一篇

高糖环境及雷帕霉素干预对肾小球足细胞Ⅳ型胶原和MMP-9表达的影响

梁馨月, 顾乐怡, 王丽华, 高嘉元, 钱家麒, 张敏芳, 戴慧莉, 严玉澄   

  1. 上海交通大学 医学院仁济医院肾脏科分子细胞（肾病）实验室, 上海 200127
  • 出版日期:2010-05-25 发布日期:2010-05-28
  • 通讯作者: 严玉澄, 电子信箱: yucheng.yan@163.com。
  • 作者简介:梁馨月（1984—）, 女, 硕士生；电子信箱: starmoonww@hotmail.com。
  • 基金资助:

    上海市重点基础研究项目（07JC14037）和上海市重大科研项目（08DZ1900500）

Effects of high glucose and Rapamycin intervention on expression of type Ⅳ collagen and matrix metalloproteinase-9 in podocytes

LIANG Xin-yue, GU Le-yi, WANG Li-hua, GAO Jia-yuan, QIAN Jia-qi, ZHANG Min-fang, DAI Hui-li, YAN Yu-cheng   

  1. Laboratory of |Molecular Cytology (Nephropathy), Department of Nephropathy, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200127, China
  • Online:2010-05-25 Published:2010-05-28
  • Supported by:

    Shanghai Key Basic Research Project, 07JC14037；Shanghai Key Scientific Research Project, 08DZ1900500

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摘要：

目的 探讨高糖培养环境及雷帕霉素干预对肾小球足细胞Ⅳ型胶原和基质金属蛋白酶9（MMP-9）表达的影响。方法 体外培养的小鼠肾小球足细胞分为空白对照组、高渗对照组、高糖组和高糖+雷帕霉素干预组。Real-time PCR和Western blotting检测各组肾小球足细胞Ⅳ型胶原α3链（COL4A3）、α5链（COL4A5）mRNA及MMP-9 mRNA和蛋白的表达；ELISA法检测细胞培养上清液中Ⅳ型胶原的含量。结果 与对照组比较，高糖组足细胞COL4A3、COL4A5 mRNA表达及培养上清液中Ⅳ型胶原含量均升高（P<0.05），而足细胞MMP-9 mRNA和蛋白表达均降低（P<0.05）。与高糖组比较，雷帕霉素干预组足细胞COL4A3、COL4A5 mRNA表达及培养上清液中Ⅳ型胶原含量均降低（P<0.05），足细胞MMP-9 mRNA和蛋白表达均升高（P<0.05）。结论 雷帕霉素对高糖环境中足细胞Ⅳ型胶原和MMP-9表达异常具有逆转作用。

关键词: 雷帕霉素, 足细胞, 高糖, Ⅳ型胶原, 基质金属蛋白酶

Abstract:

Objective To investigate the effects of high glucose and Rapamycin intervention on the expression of type Ⅳ collagen and matrix metalloproteinase-9 (MMP-9) in podocytes. Methods Mouse podocytes cultured in vitro were divided into blank control group, high permeability group, high glucose group and high glucose+Rapamycin intervention group. The expression of type Ⅳ collagen α3 chain (COL4A3) mRNA, type Ⅳ collagen α5 chain (COL4A5) mRNA, MMP-9 mRNA and MMP-9 protein in podocytes of each group was detected by Real-time PCR and Western blotting, respectively. The contents of type Ⅳ collagen in the supernatants of culture fluid were determined by ELISA. Results Compared with control group, the expression of COL4A3 mRNA and COL4A5 mRNA in podocytes and the contents of type Ⅳ collagen in the supernatants of culture fluid in high glucose group were significantly increased (P<0.05), while the expression of MMP-9 mRNA and MMP-9 protein in podocytes was significantly decreased (P<0.05). Compared with high glucose group, the expression of COL4A3 mRNA and COL4A5 mRNA in podocytes and the contents of type Ⅳ collagen in the supernatants of culture fluid in high glucose+Rapamycin intervention group were significantly decreased (P<0.05), while the expression of MMP-9 mRNA and MMP-9 protein in podocytes was significantly increased (P<0.05). Conclusion Rapamycin could reverse the abnormal expression of type Ⅳ collagen and MMP-9 in podocytes treated with high glucose.

Key words: Rapamycin, podocyte, high glucose, type Ⅳ collagen, matrix metalloproteinase