蜕皮甾酮对人表皮干细胞体外增殖的影响  

Effect of ecdysterone on proliferation of human epidermal stem cells in vitro

作　　者：李国芳[1] 吴旭[1] 付小兵[2] 张翠萍[2] 周云峰[1] 吴昌昊[1] 

机构地区：[1]南方医科大学附属南方医院胸心血管外科,广东广州510515 [2]解放军总医院第一附属医院全军烧伤研究所创伤修复重点实验室,北京100037

出　　处：《新乡医学院学报》2009年第3期246-248,F0004,共4页Journal of Xinxiang Medical University

基　　金：南方医科大学南方医院院长基金(编号:20050017)

摘　　要：目的观察蜕皮甾酮(EDS)对人表皮干细胞(hESCs)体外增殖的影响。方法采用中性蛋白酶-Ⅳ型胶原黏附法分离正常人包皮表皮干细胞,免疫细胞化学法检测第3代细胞β1整合素、角蛋白19(CK19)、角蛋白14(CK14)及角蛋白10(CK10)的表达以鉴定hESCs。分别用含0 mg.L-1(对照组)、10 mg.L-1(10 mg.L-1组)、20 mg.L-1(20 mg.L-1组)、40 mg.L-1(40 mg.L-1组)、80 mg.L-1(80 mg.L-1组)及160 mg.L-1(160 mg.L-1组)EDS的Ep ilife培养基作用第3代hESCs 3 d,四甲基偶氮唑盐(MTT)法检测酶标仪490 nm下的吸光度值,评价EDS对hESCs体外增殖的影响。结果免疫细胞化学法检测提示第3代表皮干细胞β1整合素、CK 19、CK 14的表达阳性,CK 10表达阴性。EDS作用3 d后,10 mg.L-1组、20 mg.L-1组、40 mg.L-1组、80 mg.L-1组及160 mg.L-1组吸光度均高于对照组(P<0.05);40 mg.L-1组、80 mg.L-1组和160 mg.L-1组吸光度均高于10 mg.L-1组和20 mg.L-1组(P<0.05);80 mg.L-1组吸光度高于40 mg.L-1组和160 mg.L-1组(P<0.05);其余各组间两两比较无统计学差异(P>0.05)。结论体外培养条件下EDS能促进hESCs增殖,该作用在EDS浓度为80 mg.L-1时最为显著。Objective To observe the effect of ecdysterone （EDS）on the proliferation of human epidermal stem cells （hESCs） in vitro. Methods hESCs were isolated from human foreskin by the method of protease-collagen Ⅳ-coated adhesion. At the 3rd generation of hESCs, the expression of integrins β1, keratin 19, keratin 14 and keratin 10 were examined by immunocytochemical method. After the 3rd generation of hESCs were cultured in Epilife medium containing 0 mg ·L^-1 （control group） ,10 mg ·L^-1（10 mg ·L^-1 group） ,20 mg·L^-1（20 mg ·L^-1 group） ,40 mg ·L^-1（40 mg ·L^-1 group） ,80 mg·L^-1 （80 mg· L^-1 group） or 160 mg ·L^-1（160 mg ·L^-1 group） EDS respectively for 3 days,the optieal density（OD） was measured with MTT method to evaluate the effect of EDS on proliferation of human epidermal stem cells in vitro. Results The results of immunocytochemistry showed intergrin β1 ,keratin 19 and keratin 14 were positively expressed at the 3^rd generation of hESCs, but keratin 10 was negative. OD of all experiment groups were higher than control group（ P 〈 0.05 ）. OD of 40 mg ·L^-1 group,80 mg ·L^-1 group and 160 mg ·L^-1 group were higher than that of 10 mg ·L^-1 group ang 20 mg ·L^-1 group（P 〈 0. 05）. OD of 80 mg ·L^-1 group was higher than that of 40 mg ·L^-1 group and 160 mg ·L^-1 group（ P 〈 0.05 ）. But there was nostatistically significant among other groups （ P 〉 0.05 ）. Conelusion EDS can promote the proliferation of hESCs in vitro at the optimal concentration of 80 mg ·L^-1.

关 键 词：蜕皮甾酮 表皮干细胞 增殖 体外 

分 类 号：R394[医药卫生—医学遗传学]