染料木黄酮对预防成纤维细胞光化学损伤的影响  

Preventive effect of genistein against psoralens pins ultraviolet A-induced photoaging in human dermal fibroblasts

作　　者：谭春花[1] 罗晓燕[1] 王华[1] 

机构地区：[1]重庆医科大学附属儿童医院皮肤科,400014

出　　处：《中华皮肤科杂志》2013年第4期258-261,共4页Chinese Journal of Dermatology

基　　金：2011年中华医学会一欧莱雅中国人健康皮肤/毛发研究项目（S2011-012）

摘　　要：目的探讨染料木黄酮在PuvA所致体外培养真皮成纤维细胞光老化模型中的保护作用。方法运用PuvA联合构建体外培养真皮成纤维细胞光老化模型，噻唑蓝比色法（M，ITll法）确定染料木黄酮最适光保护作用浓度，倒置显微镜下观察细胞形态，酶组织化学法观察细胞衰老相关-β-半乳糖苷酶（sA-β-Gal）表达，流式细胞仪检测细胞活性氧（R0s）含量，实时荧光定量PcR检测细胞基质金属蛋白酶1（MMP-1）mRNA表达等。结果uvA照射后24h，正常对照组、光老化组及染料木黄酮组sA-β-Gal阳性细胞率分别为（0．67±0．58）％、（96．67±1．53）％、（51．67±2．08）％，各组间差异有统计学意义（P〈0．01）。光老化组、染料木黄酮组R0s含量分别为正常对照组的（1．88±0．24）和（1．62±0．02）倍（均P〈0．01）。光老化组MMP-1mRNA表达上调为正常对照组的10倍，染料木黄酮组表达量仅为正常对照组的6倍，各组间差异有统计学意义（P〈0．05）。结论染料木黄酮对PuvA所致体外培养真皮成纤维细胞光老化具有-定的保护作用。Objective To evaluate the protective effect of genistein on psoralens plus ultraviolet A （PUVA）-indueed photoaging in human dermal fibroblasts （HDFs） in vitro. Methods Dermal fibroblasts were isolated from the foreskin of a healthy 5-year-old boy, and subjected to primary culture. After 5 - 8 passages of subculture, the fibroblasts were collected and used in the following experiment. To determine the optimal concentration of genistein, methyl thiazolyl tetrazolium （MTr） assay was conducted to detect the proliferation of fibroblasts pretreated with 8-methoxypsoralen （8-MOP） and various concentrations （0 - 20 p,g/ml） of genistein for 24 hours followed by UVA irradiation. Then, the fibroblasts were divided into 3 groups： normal control group receiving no treatment, photoaging group incubated with 8-MOP for 24 hours followed by UVA irradiation, and genistein group incubated with both 8-MOP and genistein at the optimal concentration for 24 hours followed by UVA irradiation. After additional culture, invert microscopy was carried out to observe the morphology of fibroblasts, enzyme histochemistry to assess senescent cells by using SA-I3-Gal kit, flow eytometry to determine the level of reactive oxygen species （ROS）, real-time fluorescence-based quantitative PCR to detect the matrix metalloproteinase-1 （MMP-1） expression. One-way analysis of variance was conducted to assess the differences in these parameters among these groups. Results At 24 hours after UVA irradiation, the percentage of fibroblasts positive for SA-13-galactosidase was （0.67± 0.58）%, （96.67 ±1.53）% and （51.67 ± 2.08）% in the normal control group, photoaging group and genistein group respectively, with significant differences among these groups （P 〈 0.01 ）. The level of ROS in the photoaging group and genistein group was （0.88 ±0.24） and （0.62 ± 0.02） fold higher than that in the control group（both P 〈 0.01 ）. Moreover, the MMP-1 expression level in the photoaging group and genistein gro

关 键 词：染料木黄酮 PUVA疗法 成纤维细胞 光 细胞衰老 

分 类 号：R[医药卫生]