中国血液净化

›› 2009, Vol. 8 ›› Issue (8): 441-444.

• 基础研究 • 上一篇    下一篇

辛伐他汀对炎症状态下人腹膜间皮细胞促纤维化细胞生长因子TGF-β1和CTGF表达的影响

何劲松 朱桂松   

  1. 南京大学医学院附属鼓楼医院肾脏科
  • 收稿日期:2009-02-26 修回日期:1900-01-01 出版日期:2009-08-12 发布日期:2009-08-12
  • 通讯作者: 何劲松

Effects of simvastatin on the expression of pro-fibrotic growth factors TGF-? and CTGF in human peritoneal mesothelial cells cultured in inflammatory conditions

HE Jin-song, ZHU Gui-song   

  1. Department of Nephrology, The Drum Tower Hospital Affiliated to Nanjing University Medical School, Nanjing 210008, China
  • Received:2009-02-26 Revised:1900-01-01 Online:2009-08-12 Published:2009-08-12

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摘要：

【摘要】目的 探讨辛伐他汀对体外培养的人腹膜间皮细胞在肿瘤坏死因子-(tumor necrosis factor-, TNF-)诱导后转化细胞因子-1(transforming growth factor-1, TGF-1)和结缔组织生长因子（connective tissue growth factor, CTGF）表达的影响。方法 胰蛋白酶消化法从人腹膜组织中分离间皮细胞进行原代培养并传代,分为五组(每组设3个样本)：空白对照组(完全培养液)、单纯TNF-幔1000ng/L)诱导组和TNF-(1000ng/L)诱导加低、中、高剂量辛伐他汀(2.5、5和10靘ol/L)组。采用MTT(单四唑)检测各组间皮细胞的活性；半定量RT-PCR检测细胞内TGF-1和CTGF mRNA表达； ELISA检测细胞上清液中TGF-1和CTGF的蛋白质水平，细胞沉淀用BCA蛋白检测方法测定细胞蛋白质含量，用以校正ELISA结果。结果 辛伐他汀能显著改善TNF-嵊盏己笕烁鼓ぜ淦は赴幕钚(P＜0.05, P＜0.01)，并能显著降低TNF-嵊盏己笕烁鼓ぜ淦は赴鸗GF-1和CTGF的表达(P＜0.05, P＜0.01)，两者在蛋白质和基因水平均呈剂量依赖关系。结论 辛伐他汀能抑制炎症状态下人腹膜间皮细胞TGF-1和CTGF的表达。

关键词: 辛伐他汀, 炎症状态, 人腹膜间皮细胞, 转化细胞因子-1, 结缔组织生长因子

Abstract:

【Abstract】 Objective To investigate the effects of simvastatin on the expression of TNF-α-induced TGF-β1 and CTGF in human peritoneal mesothelial cells (HPMCs). Methods HPMCs were isolated from human omenta by trypsin digestion method and cultured. HPMCs were then divided into normal control group, TNF-α-induced (1000ng/L) group and TNF-α-induced (1000ng/L) plus low-, medium- and high-dose simvastatin treated (2.5, 5 and 10μmol/L Simvastatin, respectively) groups. The viability of HPMCs was measured by MTT assay. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of TGF-β1 and CTGF mRNAs in HPMCs. TGF-β1 and CTGF in culture media were measured by enzyme linked immunosorbent assay (ELISA). Cell protein concentration was measured by trace bicinchoninic acid (BCA) method to correct the ELISA assay results. Results Low-, medium- and high-dose simvastatin significantly improved the viability of HPMCs inhibited by TNF-α (P＜0.05, P＜0.01). In addition, simvastatin significantly decreased TNF-α-induced TGF-β1 and CTGF expressions in a dose-dependent manner both at protein and mRNA levels (P＜0.05, P＜0.01). Conclusion Simvastatin inhibits the expression of TGF-β1 and CTGF in HPMCs cultured in inflammatory conditions.

Key words: inflammatory conditions, human peritoneal mesothelial cells, transforming growth factor-β1, connective tissue growth factor