吡非尼酮抑制转化生长因子β1诱导的大鼠角膜基质细胞纤维化  

Pirfenidone inhibits TGF-β1-induced fibration in rat corneal stromal cells

作　　者：吴共发[1,2] 邱丽浈[3] 黄绮亭[2] 刘钰君 曾宇婷[2] 陈俊杰[4,5] WU Gongfa, QIU Lizhen, HU-ANG Qiting , LIU Yujun, ZENG Yuting , CHEN Junjie.(Department of Pathology, Sun Yat-Sen Memorial Hospital of Sun Yat-Sen Uaiversity, Guangzhou 510120, Chin)

机构地区：[1]中山大学孙逸仙纪念医院,广州510120 [2]广州市增城区人民医院&中山大学孙逸仙纪念医院增城院区病理科,广州511300 [3]广州市增城区人民医院&中山大学孙逸仙纪念医院增城院区内三科,广州511300 [4]广州市增城区人民医院&中山大学孙逸仙纪念医院增城院区眼科,广州5113005 [5]广西贺州光明眼科医院,广西贺州542800

出　　处：《实用医学杂志》2018年第12期1955-1958,共4页The Journal of Practical Medicine

基　　金：广东省医学科研基金项目(编号:A2016422);广州市增城区人民医院优秀医学人才培育基金项目(编号:2017-YX-001)

摘　　要：目的观察吡非尼酮(PFD)对转化生长因子β1(TGF-β1)诱导的大鼠角膜基质细胞向成纤维细胞转化过程的影响,探讨PFD抗角膜基质纤维化的作用。方法分离培养大鼠角膜基质细胞,免疫细胞化学法检测波形蛋白(Vimentin)进行细胞鉴定。实验分为对照组(DMEM+10%FBS)、TGF-β1组(2 ng/mL TGF-β1+DMEM+10%FBS)和PFD组(1 mg/mL PFD+2 ng/mL TGF-β1+DMEM+10%FBS)。CCK-8法检测细胞增殖能力,Western blot检测细胞CollagenⅠ、CollagenⅢ、Keratocan和CD90蛋白表达。结果大鼠角膜基质细胞呈Vimentin胞浆阳性。与对照组及TGF-β1组比较,PFD组细胞的增殖OD值显著减低(P<0.05)。Western blot显示PFD组的细胞CollagenⅠ、Keratocan蛋白表达增强而CollagenⅢ和CD90蛋白表达减弱(P<0.05)。PFD组的CollagenⅢ/CollagenⅠ比值在3组中最低(P<0.05)。结论 PFD抗角膜基质细胞纤维化是通过抑制TGF-β分子途径,从而影响胶原合成和Keratocan、CD90蛋白表达实现的。Objective To study the effect of pirfenidone（PFD）on the transformation of rat cornealstromal cells into fibroblasts in vitro and further explore the anti-fibrotic effect of PFD. Methods The cornealstromal cells from SD rat was isolated and cultured,and was determined by vimentin stain. The experiment wasdivided into control group（DMEM ＋ 10%FBS）,TGF-β1 group（2 ng/mL TGF-β1＋ DMEM ＋ 10%FBS）and PFDgroup（1 mg/mL PFD ＋ 2 ng/mL TGF-β1 ＋ DMEM ＋ 10%FBS）. Cell proliferation was detected by CCK-8 assay.Collagen Ⅰ,Collagen Ⅲ,Keratocan and CD99 expression were detected by Western blot. Results Comparedwith control group and TGF-β1 group,the cell proliferation were significantly decreased in PFD group（P〈0.05）.Western blot showed that PFD can up-regulated Collagen Ⅰ and Keratocan but down-regulated Collagen Ⅲ CD90 expression（P〈0.05）. The ratio of Collagen Ⅲ/Collagen Ⅰ in PFD group was lowest in all groups（P〈0.05）.Conclusion PFD can resistant fibration in corneal stromal cells may through the inhibition of TGF-β,whichaffect the collagen synthesis and Keratocan,CD90 expression.

关 键 词：吡非尼酮 角膜基质细胞 转化生长因子Β1 纤维化 

分 类 号：R965[医药卫生—药理学]