蛋白激酶C-ε对人胚肺成纤维细胞趋化运动调节的研究  

Protein kinase C-ε mediates chemotaxis of human fetal lung fibroblasts

作　　者：方秋红[1] 马迎民[1] 刘相德[2] Stethen I.Rennard 

机构地区：[1]北京世纪坛医院呼吸科,北京100038 [2]美国内布拉斯加大学医学中心呼吸内科,内布拉斯加州奥马哈

出　　处：《中国实用内科杂志》2008年第4期264-266,共3页Chinese Journal of Practical Internal Medicine

基　　金：国家自然科学基金(30570791)

摘　　要：目的探讨蛋白激酶C-ε(PKC-ε)家族在肺成纤维细胞向纤维粘连蛋白趋化运动中的作用。方法于2006年10月至2007年4月在北京世纪坛医院内科实验室和美国内布拉斯加大学呼吸科实验室进行。(1)人胚肺成纤维细胞体外培养。(2)以纤维粘连蛋白作为趋化因子,采用Boyden趋化小室技术测定人胚肺成纤维细胞的趋化运动。(3)将人胚肺成纤维细胞转染PKC-ε小干扰RNA(siRNA),24h后应用蛋白质印迹法测定PKC-ε的表达。(4)分别采用PKC-ε非选择性抑制剂(CalphostinC)和选择性抑制剂(Ro-31-8220)及转染PKC-εsiRNA的人胚肺成纤维细胞,研究PKC-ε在肺成纤维细胞趋化运动中的调节作用。结果CalphostinC和Ro-31-8220显著抑制人胚肺成纤维细胞向纤维粘连蛋白的趋化运动[细胞迁移数分别为对照组的(68.50±4.68)%和(59.94±5.20)%,P<0.01]。成纤维细胞转染PKC-εsiRNA,孵育24h后PKC-ε的表达显著减少,成纤维细胞在纤维粘连蛋白作用下的趋化运动显著减弱[迁移的细胞数是对照组的(82.56±2.80)%,P<0.05)]。结论PKC-ε参与肺成纤维细胞趋化运动的调节,进而在肺成纤维细胞介导的组织重构中发挥作用。Objective To investigate the effect of protein kinase C-ε （PKC-ε） on chemotaxis of human fetal lung fibroblasts（HFL） induced by fibronectin. Methods The study was performed in Beijing Shijitan Hospital and American Nebraska Medical Center from October 2006 to April 2007. HFL were cultured in 100-mm tissue culture dishes in Dulbecco＇s modified Eagle＇s medium （DMEM）supplemented with 10% FCS, 50 U/mL penicillin, 50μg/mL streptomycin, and 1μg/mL fungizone. Boyden blindwell chamber method was utilized to measure chemotaxis of HFL. PKC-ε knockdown in HFL was obtained by transfection of small interfering RNA （siRNA）targeting human PKC-ε. The efficacy of knockdown was assessed by Western blot. Results Chemotaxis of HFL was significantly inhibited by both non-selective and selective PKC-ε inhibitors. PKC-ε knockdown was obviously observed after transfection of PKC-ε siRNA. With knockdown of PKC-ε,chemotaxis of HFL was decreased. Conclusion Fibronectin-driven chemotaxis of HFL was at least partially mediated by PKC-ε.

关 键 词：成纤维细胞 蛋白激酶C-ε 纤维粘连蛋白 

分 类 号：R5[医药卫生—内科学]