肝细胞生长因子对正常皮肤和增生性瘢痕成纤维细胞的影响  

Effects of hepatocyte growth factor on fibroblast of normal skin and hypertrophic scar

作　　者：李金凤[1] 段海峰[1] 吴彬[1] 毕建进[1] 孙同柱[2] 付小兵[2] 王立生[1] 吴祖泽[1] 

机构地区：[1]军事医学科学院放射与辐射研究所实验血液学实验室,北京100850 [2]解放军总医院第一附属医院全军创伤修复重点实验室,北京100037

出　　处：《感染．炎症．修复》2007年第3期140-143,F0002,共5页Infection Inflammation Repair

基　　金：国家重点基础研究发展规划973项目(2004CB518801)

摘　　要：目的:采用携带肝细胞生长因子(HGF)腺病毒转染正常皮肤成纤维细胞(NFB)和增生性瘢痕成纤维细胞(HFB)后,观察HGF对这两种细胞作用的差异,为HGF基因治疗病理性瘢痕提供理论基础。方法:分离培养HFB和NFB,以不同感染复数(multiplicity of infection,MOI)携带绿色荧光蛋白基因的腺病毒(Ad-GFP)转染细胞,用流式细胞仪检测转染效率;以MOI为100携带HGF基因的腺病毒(Ad-HGF)转染HFB和NFB48h后,酶联免疫吸附法检测细胞上清中HGF、基质金属蛋白酶-1(MMP-1)和转化生长因子-β1(TGF-β1)的表达;应用免疫组织化学方法检测正常皮肤和增生性瘢痕组织中MMP-1和TGF-β1的分布。结果:NFB和HFB在腺病毒转染效率和HGF表达方面差异无显著性,但HGF能明显促进HFB分泌MMP-1,抑制HFB表达TGF-β1。结论:促进HFB分泌MMP-1和抑制HFB表达TGF-β1可能是HGF治疗病理性瘢痕的细胞和分子生物学基础。Objective：To explore the differences between the influences of hepatoeyte growth factor （HGF） on the fibroblasts of normal skin and that of hypertrophic scar tissue with the aid of transfection of the recombinant adenovirus carrying HGF gene （Ad-HGF）. Methods： Primary cultured fibroblasts from normal skin and hypertrophic scar were transfected with the recombinant adenovirus carrying green fluorescence protein gene （Ad-GFP） at different multiplicity of infection （MOI）, and the transfection efficiencies were evaluated by flow cytometry. Levels of HGF, matrix metalloprotease-1 （MMP-1） and TGF-β1 in cell culture medium were determined by enzyme linked immunoadsorbent assay （ELISA） after the fibroblasts were transfected with Ad-HGF at 100 MOI. Immunohistochemistry was used to determine the expression of MMP-1 and TGF-β1 in normal skin and hypertrophic scar. Results：There was no significant difference from the transfection efficiencies and the expressions of HGF between the two fibroblasts. HGF could obviously promote the expression of MMP-1 in HFB, and inhibit the secretion of TGF-β1 in HFB only. The results of immunohistochemistry revealed that the difference in the expression level of MMP-1 and TGF-β1 between normal skin and hypertrophic scar was distinct. Conclusion： The mechanism underlying the efficacy of HGF in treatment of hypertrophic scar may be associated with the ability of HGF to inhibit TGF-β1 expression and promote MMP-1 secretion in hypertrophic scar fibroblasts.

关 键 词：肝细胞生长因子 增生性瘢痕 成纤维细胞 腺病毒 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]