吡非尼酮对人椎板切除术后硬膜外瘢痕细胞的作用及机制研究  

Effects of pirfenidone on epidural scar cells after laminectomy and related mechanisms

作　　者：王凤珍 王兆红[2] 夏计划[2] 周恒才[2] 拾坤[2] WANG Fengzhen;WANG Zhaohong;XIA Jihua;ZHOU Hengcai;SHI Kun(Department of Clinical Pharmacy,School of the First Clinical Medicine,Xuzhou Medical University,Xuzhou,Jiangsu 221002,China;Department of Orthopedics,Xuzhou Central Hospital,Xuzhou,Jiangsu 221009)

机构地区：[1]徐州医科大学第一临床学院临床药学教研室,江苏徐州221002 [2]徐州市中心医院骨脊柱外科,江苏徐州221009

出　　处：《徐州医科大学学报》2020年第11期795-799,共5页Journal of Xuzhou Medical University

基　　金：徐州市应用基础研究计划(KC-18041,KC-19069);国家自然科学基金青年基金(81803473);江苏省药学会基金(Q2018051);北京公益基金(YWJKJJHKYJJ-A720)。

摘　　要：目的探讨吡非尼酮对人椎板切除术后硬膜外瘢痕细胞的作用及机制。方法采用体外组织块培养人硬膜外瘢痕细胞并建立细胞系,分别加入不同浓度的吡非尼酮培养,CCK-8法检测细胞活力,细胞划痕法观察细胞迁移,TUNEL法检测细胞凋亡,Western blot分析转化生长因子β1(TGF-β1)、α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原蛋白、Smad2、磷酸化的Smad2(phospho-Smad2)、Smad3、磷酸化的Smad3(phospho-Smad3)表达。结果成功分离培养人硬膜外瘢痕成纤维细胞。吡非尼酮可抑制成纤维细胞的活性(P<0.05),且并未增加成纤维细胞的凋亡(P>0.05)。吡非尼酮可抑制细胞的迁移,并抑制α-SMA、Ⅰ型胶原蛋白表达及细胞增殖相关蛋白的表达和磷酸化。结论吡非尼酮可抑制瘢痕成纤维细胞的活性及迁移,其机制可能是通过抑制TGF-β/Smad通路及肌成纤维细胞转化实现的。Objective To investigate the effects of pirfenidone on epidural scar cells after laminectomy and related mechanism.Methods The human epidural scar cells were cultured in vitro.Then,the cells were exposed to different concentrations of pirfenidone.Their viability was measured by CCK-8 assay.Their migration was measured by the wound healing assay.The cell apoptosis was detected by in situ terminal transferase labeling(TUNEL).The levels of transfer growth factorβ1(TGF-β1),α-SMA,collagen type I,Smad2,phospho-Smad2,Smad3 and phospho-Smad3 were measured by Western blot.Results Human epidural scar fibroblasts were successfully isolated and cultured.Pirfenidone decreased the viability of fibroblasts(P<0.05),but did not stimnlate increase the apoptosis of fibroblasts(P>0.05).Pirfenidone inhibited cell migration and the expression ofα-SMA,collagen type I,cell proliferation-related proteins and proteins phosphorylation.Conclusions Pirfenidone inhibits the viability and migration of fibroblasts,which may be associated with inhibition of the TGF-β/Smad pathway and myofibroblast transformation.

关 键 词：吡非尼酮 成纤维细胞 迁移 增殖 分子机制 

分 类 号：R681.5[医药卫生—骨科学]