吡非尼酮对转化生长因子-β_1诱导的大鼠心肌成纤维细胞增殖和胶原蛋白合成的影响  

Effect of Pirfenidone on TGF-β_1 induced proliferation and collagen synthesis of rat cardiac fibroblasts

作　　者：王琳娜[1] 陈常勇[2] 郭存霞[1] 陈小永[1] 陈香美[3] 

机构地区：[1]河南省中医院肾病科,河南郑州450002 [2]中南大学湘雅医院放射科,湖南长沙410008 [3]中国人民解放军总医院肾脏病科(肾脏疾病国家重点实验室),北京100853

出　　处：《中国现代医学杂志》2015年第29期30-34,共5页China Journal of Modern Medicine

基　　金：河南省中医药科学研究专项基金(No:2014ZY02021)

摘　　要：目的探讨吡非尼酮(PFD)对转化生长因子-β1(TGF-β1)诱导的乳鼠心肌成纤维细胞(CFB)增殖和Ⅰ型胶原蛋白(ColⅠ)合成的影响,探讨其抗纤维化作用机制。方法 1体外培养CFB,不同浓度(0、100、200、400、800和1 600μg/ml)PFD加入CFB培养液中24、48和72 h,四甲基偶氮唑盐(MTT)比色法检测CFB增殖,筛选PFD最佳作用浓度范围和作用时间;2根据以上实验结果,选用400和1 600μg/ml PFD加入CFB培养液中48 h作为观察浓度和干预时间,实时定量逆转录-聚合酶链反应(RT-q PCR)和Western blot检测PFD对TGF-β1诱导CFB增殖、ColⅠm RNA和ColⅠ表达的影响。结果与对照组比较,400和1 600μg/ml PFD能显著抑制TGF-β1诱导CFB增殖、ColⅠm RNA和ColⅠ表达,400μg/ml即能起显著抑制作用。结论 PFD对CFB增殖和胶原蛋白合成有抑制作用,PFD的抗纤维化作用与其有关。【Objective】 To investigate the effect of Pirfenidone（PFD） on transforming growth factor-β1（TGF-β1） induced proliferation and type Ⅰcollagen（Col Ⅰ） synthesis in rat cardiac fibroblasts（CFB） and explore its anti-fibrotic mechanism. 【Methods】 Cardiac fibroblasts were treated with PFD of different concentrations（0, 100, 200, 400, 800 and 1,600 μg/ml） for 24, 48 and 72 hours. Cell proliferation activity was detected by MTT assay and then the appropriate concentration and time of Pirfenidone were found out.The cultured cardiac fibroblasts were stimulated with TGF-β1and then treated with PFD of different concentrations（400 and 1,600 μg/ml） for 48 hours. Cell proliferation activity was detected by MTT assay, the expression of type Ⅰ collagen m RNA was determined by real-time quantitative reverse transcription-polymerase chain reaction（RT-q PCR） and the amount of type Ⅰ collagen in cultured cells was measured by Western bolt. 【Results】 Pirfenidone inhibited cardiac fibroblast proliferation stimulated with TGF-β1, the expression of type Ⅰ collagen m RNA and the amount of type Ⅰ collagen. The effective concentration of Pirfenidone was 400 ~ 1,600 μg/ml for 48 h after Pirfenidone was added into culture medium, the concentration of 400 μg/ml worked significantly. 【Conclusion】 Pirfenidone can inhibit cardiac fibroblast proliferation stimulated with TGF-β1and collagen production which may contribute to its anti-fibrotic mechanism.

关 键 词：吡非尼酮 心肌成纤维细胞 尿毒症 Ⅰ型胶原蛋白 转化生长因子-Β1 

分 类 号：R542.23[医药卫生—心血管疾病] R96[医药卫生—内科学]