GX-50对中波紫外线辐射皮肤成纤维细胞的保护作用  

Rotective Effects of GX-50 on Ultraviolet-B Exposed Skin Fibroblasts

作　　者：高萍[1] 宋小飞 戴靖波[1] 王朝霞[1] 徐汪节[1] 乔中东[1] 

机构地区：[1]上海交通大学生命科学技术学院,上海200240 [2]上海瑞金集团闵行中心医院泌尿外科,上海201100

出　　处：《上海交通大学学报（农业科学版）》2015年第2期47-53,共7页Journal of Shanghai Jiaotong University(Agricultural Science)

基　　金：江苏省无锡530计划(2011JDZX020)

摘　　要：为观察花椒中的提取物GX-50是否对UVB诱导的光老化皮肤细胞具保护作用,使用组织块法分离培养原代人皮肤成纤维细胞,通过观察不同剂量UVB照射后的细胞形态学改变确定后续实验的UVB单次照射剂量;MTT法检测不同浓度GX-50处理后的细胞活性;采用衰老相关β-半乳糖苷酶(SA-β-Gal)细胞化学染色法确定皮肤细胞衰老程度;测定丙二醛(MDA)和ROS含量评估皮肤细胞氧化损伤的程度;酶联免疫吸附试验(ELISA)检测细胞TGF-β1表达量的变化;Western blot法检测细胞胶原蛋白表达量的变化。结果显示UVB辐射成纤维细胞后,细胞胞体增大、颗粒增多、部分细胞死亡,在不引起细胞明显死亡的UVB最大剂量50 mJ/cm2作用下,10-6mol/L的GX-50在孵育24h时可显著增加成纤维细胞存活率(125.7%,P<0.01);经UVB照射后细胞SA-β-Gal染色阳性率增加,GX-50预处理后辐射组的阳性率明显降低(45.9%,P<0.05);UVB组与对照组相比,MDA、ROS含量明显上升,GX-50预处理后辐射组MDA、ROS含量分别下降了22.6%(P<0.01),22.8%(P>0.05),基本与对照组持平;UVB辐射后TGF-β1分泌减少,GX-50预处理后辐射组则明显上升(14.7%,P<0.05);Western blot结果显示,UVB可显著抑制胶原蛋白的表达,而GX-50预处理后表达量则基本恢复。推断GX-50对UVB引起的皮肤成纤维细胞损伤起到了保护作用,其机制可能是清除自由基和抗氧化,同时活化被UVB抑制的TGF-β信号通路,从而增加了胶原蛋白的表达。To clarify the protective effect of GX-50 on skin fibroblasts under UVB irradiation and the mechanism underlying it,primary human skin fibroblasts were isolated and cultured by tissue explants method,and UVB irradiation dose for subsequent experiments was determined through observation of cell morphology changes after irradiation with different doses of UVB;the effect of GX-50 on skin fibroblasts viability was determined through MTT tests;the anti-aging effect of GX-50 was determined by senescenceassociated-β-galactosidase（SA-β-Gal）staining on skin fibroblasts.Secreted malondialdehyde（MDA）concentration and intracellular ROS in fibroblast culture were determined to evaluate the extent of the oxidative damage;the expression of TGF-β1was detected by ELISA,and the expression of collagen in cells was detected by Western blot.After UVB radiation,the cell body increases,the particles within cells become more and cell apoptosis appears.Under the largest UVB dose,50 mJ/cm^2,which did not induce obvious cell death,10-6 M GX-50 significantly increased cell viability compared to non-treated cells（125.7%,P〈0.01）;the SAβ-gal positive cells of UVB-irradiated group were higher than those of nonirradiated group and GX-50 significantly inhibited theβ-galactosidase activity（45.9%,P〈0.05）;pretreatment with GX-50 attenuated an increase in intracellular ROS（22.8%,P〉0.05）and MDA content（22.6%,P〈0.01）,which was triggered by UVB irradiation;there was a significant decrease of the expression of TGF-β1after UVB irradiation and GX-50 pretreatment was found to increase it（14.7%,P〈0.05）;UVB can significantly inhibit collagen expression,which basically recovered after pretreatment with GX-50.Overall,these results present evidence to explore the protective effect of GX-50 against senescencelike characteristics induced by UVB on fibroblasts.The mechanism underlying it may be the antioxidative action or activation of the TGF-βsignal pathways,which is inhibited by UVB,thus increasing the collagen expres

关 键 词：GX-50 皮肤老化 中博紫外线(UVB) 成纤维细胞 

分 类 号：R758.14[医药卫生—皮肤病学与性病学]