FADD-DN基因转梁对氦氖激光诱导瘢痕成纤维细胞凋亡的影响

摘要

Aim To explore the molecular mechanism of He-Ne laser-inducing apoptosis of scar fibroblasts in vitro.Methods These fibroblasts-devired from hypertrophic scar (HS) were cultured and were divided into three groups:transfection group (TG),control transfection group (CTG) and non-transfection group (NTG).The fibroblasts in TG were co- transfected with GFP (green fluorescent protein) cDNA and pcDNA3-FADD-DN. Scar fibroblasts in CTG were cotransfected with GFP cDNA and pcDNA3. The fibroblasts in NTG was't transfected. Scar fibroblasts of each group were irradiated with He-Ne laser (100 mW/cm2 for 30 minutes) at 24 hours after transfection, once a day for 3 consecutive days.Apoptosis of cell was then examined by TUNEL technique. Results The percent of apoptosis in TG was 9.27% and that of apoptosis was 15.73% in CTG, and 12.87% in NTG.The percent of apoptosis in TG was less than in CTG (q=4.55,P< 0.01) and NTG(q=4.14,P< 0.05) respectively.Conclusion FADD-DN gene transfection can depress He-Ne laser-inducing apoptosis of scar fibroblasts.FADD may be an essential component of the He-Ne laser-activited cell death pathway.%目的探讨氦氖激光诱导瘢痕成纤维细胞凋亡的分子机制.方法将培养瘢痕成纤维细胞随机分为转染组、转染对照组和非转染组,转染组细胞与绿色荧光蛋白 (GFP)质粒 cDNA、含目的基因片段的表达载体质粒 pcDNA3-FADD-DNcDNA共转染,转染对照组细胞与 GFP质粒 cDNA、空载体质粒 pcDNA3共转染,非转染组细胞没有进行转染,转染后 24 h分别对各组细胞进行氦氖激光照射( 100 mW/cm2照射 30 min), 1次 /d,连续照射 3 d,然后采用 TUNEL技术检测细胞凋亡.结果转染组、转染对照组与非转染组细胞凋亡率分别为 9.27% ,15.73%和 12.87%,转染组细胞凋亡率明显小于转染对照组 (q=4.55, P< 0.01)和非转染组( q=4.14, P< 0.05).结论 FADD-DN基因转染能减少氦氖激光诱导的瘢痕成纤维细胞凋亡, FADD可能是氦氖激光激活死亡信号途径的基本元件之一.