黄芩苷抑制长波紫外线诱导的人成纤维细胞氧化损伤和凋亡  

Baicalin inhibits ultraviolet A-induced oxidative damage and apoptosis in human fibroblasts

作　　者：尹慧彬[1] 周炳荣[1] 骆丹[1] 吴维[1] 方晓波[1] 张倩[1] 郭泽[1] 张家安[1] 胡燕燕[1] 

机构地区：[1]南京医科大学第一附属医院皮肤性病科,210029

出　　处：《中华皮肤科杂志》2013年第4期266-269,共4页Chinese Journal of Dermatology

基　　金：国家自然科学基金（81000700.81171518）

摘　　要：目的探讨黄芩苷对长波紫外线（UVA）诱导的人二倍体成纤维细胞（HDF）氧化损伤和凋亡的抑制作用及其可能机制。方法CCK-8法检测细胞活性，筛选最佳药物浓度。将成纤维细胞分成对照组、UVA照射组、UVA＋6．25mg／L黄芩苷组、UVA＋12．5mg／L黄芩苷组、UVA＋25m∥L黄芩苷组。利用荧光显微镜和流式细胞仪检测各组细胞中活性氧、超氧化物阴离子和一氧化氮含量的变化；流式细胞仪检测线粒体膜电位和细胞凋亡率的变化。Western印迹法检测凋亡相关蛋白caspase3含量。以上多组间比较均采用单因素方差分析。结果UVA照射组的活性氧、超氧化物阴离子和一氧化氮含量分别为813．1±30．29、353．3±19．13、107．1±11．44，较对照组（442．5±29．45、223．4±10．67、42．17±2．59）明显上升，均P〈0．05）；同时，UVA照射组较对照组线粒体去极化增强，凋亡率上升。与UVA组比较，各浓度黄芩苷作用组氧化产物包括活性氧、超氧化物阴离子及一氧化氮含量均有明显下降，线粒体去极化减弱，细胞凋亡率也显著下降，均P〈0．01。UVA组凋亡相关蛋白caspase3相对含量为1．2680±0．0091，明显高于对照组（0．3675±0．1115），6．25、12．5和25mg／L黄芩苷组（0．5588±0．1705、0．5365±0．1718、0．5454±0．2083）较UVA组有显著下调（均P〈0．01）。结论黄芩苷可以减轻UVA诱导的人成纤维细胞氧化损伤和细胞凋亡，其机制可能与清除活性基团、抑制线粒体膜去极化、阻止下游凋亡蛋白caspase3的激活和表达有关。Objective To investigate the inhibitory effect of baicalin on ultraviolet A （UVA）-induced oxidative damage and apoptosis in human diploid fibroblasts （HDFs） and its possible mechanism. Methods Diploid fibroblasts were isolated from healthy aduh male foreskin tissue and subjected to primary culture and subculture. To determine the working concentrations of baicalin, CCK-8 assay was performed to evaluate the proliferation of HDFs treated with various concentrations （3.125-100 mg/L） of baicalin alone or in combination with UVA irradiation. Then, some HDFs were divided into 5 groups： the control group receiving no treatment, UVA group irradiated with UVA alone, 3 combination groups irradiated with UVA followed by treatment with baicalin at 6.25 mg/L, 12.5 mg/L and 25 mg/L, respectively. After additional cuhure for 24 hours, fluorescence microscopy and flow cytometry were performed to detect and quantify reactive oxygen species, superoxide anion and nitric oxide, flow cytometry to determine mitoehondrial membrane potentials and cell apoptosis rate, and Western blot to measure the expression of caspase 3. One way analysis of variance was carried out to compare these parameters among these groups. Results Compared with the control group, the UVA group showed a significantly higher level of reactive oxygen species, superoxide anion and nitric oxide （813.1 ± 30.29 vs. 442.5 ± 29.45,353.3 ± 19.13 vs. 223.4 ± 10.67,107.1 ± 11.44 vs. 42.17 ± 2.59, all P 〈 0.05）, together with elevated mitochondrial membrane potential （771.6 ± 68.27 vs. 429.5 ±27.14, P〈 0.05） and apoptosis rate （18.14 ± 2.492 vs. 2.90 ± 0.574, P 〈 0.05）. There was a significant decrease in the level of reactive oxygen species, superoxide anion and nitric oxide, mitochondrial membrane potential and apoptosis rate in the 3 combination groups compared with the UVA group （all P 〈 0.01 ）. The relative expression level of caspase-3 was significantly higher in the UVA group than in the control group （1.2680 ± 0.0091

关 键 词：紫外线 成纤维细胞 黄芩甙 氧化性应激 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]