肌动蛋白重构蛋白对瘢痕成纤维细胞增殖和分化的影响  

Study of the effect of actin remodeling protein on proliferation and differentiation of fibroblast car hypertrophic scar

作　　者：贾小丽[1] 高锋[1] 官菊梅[2] 

机构地区：[1]四川中医药高等专科学校附属医院皮肤科,四川绵阳621000 [2]成都中医药大学基础医学院,成都610075

出　　处：《重庆医学》2015年第17期2347-2350,共4页Chongqing medicine

基　　金：教育部科学技术研究重点项目(211158)

摘　　要：目的观察肌动蛋白重构蛋白(flightlessⅠ,FliⅠ)对人瘢痕成纤维细胞增殖和分化的影响,探讨瘢痕组织形成的部分机制。方法将实验分为空白组、Lipo2000组、FliⅠsiRNA组、FliⅠcDNA组,采用WST-1法检测细胞增殖,RT-PCR法、Western blot法分别检测FliⅠ、COL-1mRNA及其蛋白的表达水平。结果转染48h后,各组成纤维细胞平均吸光度(A)值分别为:空白组0.934±0.153,Lipo2000组0.910±0.084,FliⅠsiRNA组1.330±0.216,FliⅠcDNA组0.607±0.042,FliⅠsiRNA组细胞增殖水平明显高于其他3组(P<0.05);FliⅠcDNA组细胞增殖水平明显低于其他3组(P<0.05);RT-PCR法检测FliⅠ、COL-1mRNA的表达:与空白组比较,FLiⅠsiRNA组扩增条带亮度减弱,FLiⅠcDNA组扩增条带亮度明显增强,FLiⅠsiRNA组的FliⅠ与COL-1扩增产物条带相对灰度值较空白组分别降低至0.194±0.023和0.316±0.176,均显著低于其他3组(P<0.01),而FLiⅠcDNA组的FliⅠ与COL-1扩增产物条带相对灰度值较空白组分别升高至7.846±1.075和9.842±1.659,均显著高于其他3组(P<0.01);与空白组及Lipo2000组比较,FLiⅠsiRNA组的成纤维细胞中FliⅠ、COL-1蛋白表达显著减弱(P<0.05,P<0.01),FLiⅠcDNA组细胞中FliⅠ、COL-1蛋白表达显著增强(P<0.05,P<0.01)。结论 FLiⅠ缺失能增强瘢痕成纤维细胞的增殖,并能在转录和翻译水平下调FLiⅠ与COL-1的表达,可能是瘢痕增生的机制之一。Objective To study the role of FLiⅠ on the proliferation and differentiation of human fibroblast hypertrophic scar and part of the mechanism the scar formation.Methods All cases were divided into 4groups,control group,Lipo2000 group,FliⅠ SiRNA group,FliⅠ cDNA group.Cell proliferation was performed by WST-1,and FliⅠ mRNA expression,COL-1 mRNA and its protein level were detected by assay to detect by RT-PCR and Western blot.Results After 48 hof transfection,the average value of absorbance in different groups were as follows：the control group was 0.934±0.153,Lipo2000 control group was 0.910±0.084,FliⅠ siRNA group was 1.330±0.216,FliⅠ cDNA group was 0.607±0.042,the level of cell proliferation in FliⅠ siRNA group was significantly higher than those in the other 3groups（P〈0.05）,while the FliⅠ cDNA group was significantly lower than those in the other 3groups（P〈0.05）;FliⅠ mRNA,COL-1mRNA expression by RT-PCR assay.Compared with the control group,FLiⅠ siRNA group amplified bands brightness decreases,FliⅠ cDNA amplification bands set the brightness significantly enhanced,FliⅠ and COL-1amplification products relative gray bands value of FliⅠ siRNA group was significantly lower than those in the other 3groups（P〈0.05）,while FliⅠ and COL-1amplification products relative gray bands value of FliⅠ cDNA group（7.846 ± 1.075 and 9.842 ± 0.659）was significantly higher than those in the other 3groups（P〈0.05）;FliⅠ and COL-1protein levels by Western blot.Compared with the control group and Lipo2000 group,the expression of FliⅠ and COL-1protein in FLiⅠ siRNA group significantly decreased（P〈0.05,P〈0.01）,while FLiⅠ cDNA group significantly increased（P〈0.05,P〈0.01）.Conclusion FliⅠ deficiency could enhance the proliferation of fibroblasts,and could low down the FliⅠ and COL-1gene expression at transcriptional and translational level,this may be one of mechanism of scar formation.

关 键 词：瘢痕 成纤维细胞 肌动蛋白重构蛋白 胶原Ⅰ型 

分 类 号：R622[医药卫生—整形外科]