波长585nm脉冲染料激光对成纤维细胞增殖以及其前胶原基因mRNA表达的影响

Effects of pulsed dye laser on the procollagen mRNA expression and proliferation of cultured fibroblasts in vitro.

以不同能量密度的脉冲染料激光照射体外培养的正常皮肤的成纤维细胞,照射后继续培养24h,分别以MTT法检测其增殖功能,以定量PCR(LightCycler)法检测其细胞内不同类型的前胶原基因及SMADs mRNA的表达.结果显示能量密度为5 J/cm2和7 J/cm2的脉冲染料激光能抑制体外培养的正常皮肤的成纤维细胞的增殖功能(P<0.05),在能量密度为5 J/cm2时能显著抑制I型前胶原基因1α(P<0.01)以及SMAD 2,3,4,7 mRNA的表达(P<0.05或P<0.01),下调I型前胶原基因1α与III型前胶原基因mRNA的比率.在能量密度为6,7 J/cm2时能抑制III型前胶原基因α1mRNA的表达(P<0.05),而对I型前胶原基因1α,2α的表达与对照无显著性差异.提示波长585 nm脉冲染料激光对成纤维细胞的增殖及其胶原产生的某些相关基因的表达具有直接的作用,且与能量密度有关.

Cultured fibroblasts were exposed to 585-nm flashlamp-pumped pulsed dye laser （ fluence 3 J/ cm^2,5 J/cm^2, 7 J/cm^2, spot size 7 mm, pulse duration 450 μs ） followed by 24 h incubation. The proliferation of fibroblasts was measured by MTT methods and the expression of the procollagen mRNA and SMADs was investigated by real time PCR method. The results showed that 585 nm flashlamp- pumped pulsed dye laser inhibited the proliferation of fibroblasts after exposure to fluence of 5 J/cm^2 and 7 J/cm^2 （P〈 0.05）. The mRNA expression of procollagen Iαl and SMAD2, 3, 4, 7 was significantly down-regulated after being treated with 585 nm flashlamp-pumped pulsed dye laser at fluence level of 5 J/cm^2（P〈0.01, or P〈0.05）. The corresponding fluence level for significant down-regulation of mRNA expression of procollage Ⅲ α1 was 6 J/cm^2 and 7 J/cm^2（P〈 0.05）. It is concluded that sufficiently high level fluence of 585 nm flashlamp-pumped pulsed dye laser may inhibit the proliferation of fibroblasts in dose-dependent manner in vitro and down- regulate procollagen mRNA expression.