白癜风皮损旁黑素细胞自噬相关蛋白表达的研究  

Expressions of autophagy-related proteins in perilesional melanocytes of vitiligo in vitro

作　　者：虞海燕[1] 岑建萍[1] 林晓霞[2] 何剑萍[1] 王瑶瑶 程浩[1] YU Haiyan;CEN Jianping;LIN Xiaoxia(Department of Dermatology,Sir Run Run Shaw Hospital,Zhejiang University School of Medicine,Hangzhou 310016,China)

机构地区：[1]浙江大学医学院附属邵逸夫医院皮肤科,杭州310016 [2]浙江大学医学院临床医学研究所

出　　处：《浙江医学》2019年第24期2592-2595,共4页Zhejiang Medical Journal

基　　金：浙江省自然科学基金资助项目（LY16H110001,LY15H110001)

摘　　要：目的通过比较体外培养的白癜风黑素细胞与正常黑素细胞中自噬标志物微管相关蛋白轻链3(LC3II/I)、自噬相关蛋白p62、哺乳动物雷帕霉素靶蛋白(m TOR)和自噬相关基因5(Atg5)的表达差异,分析其在白癜风发病中的可能作用。方法分离并原代培养来自于泛发型白癜风患者皮损旁皮肤的黑素细胞(白癜风黑素细胞组)以及正常志愿者皮肤的黑素细胞(正常对照组),采用免疫印迹法分别检测不同来源黑素细胞中自噬标志物LC3II/I、自噬相关蛋白p62、m TOR和Atg5的表达,同时以改良的多巴氧化法测定细胞中的酪氨酸酶活性。结果LC3II/I及Atg5的表达在白癜风黑素细胞组中明显高于正常对照组(t=10.36、13.01,均P<0.01),而p62、m TOR蛋白水平及酪氨酸酶活性在白癜风黑素细胞组中明显低于正常对照组(t=5.12、7.22、36.51,均P<0.01)。结论体外培养的白癜风黑素细胞存在自噬的活化及上调并伴有黑素细胞黑素合成功能的下降,提示自噬可能与白癜风的发病有关。Objective To investigate the expressions of autophagy markers microtubule-associated protein-1 light chain 3(LC3 II/I and autophagy related protein P62,mammalian target of rapamycin(m TOR)and autophagy related gene 5(Atg5)in vitiligo melanocytes and normal melanocyts.Methods Primary cultured melanocytes were isolated from perilesional skin of patients with generalized vitiligo(vitiligo melanocytes)or skin of normal subjects(normal controls melanocytes).The expressions of autophagy markers LC3 II/I and autophagy related protein P62,m TOR and Atg5 in different sources of melanocytes were determined by Western blot,and the tyrosinase activity in melanocytes were measured using the modified dopachrome method.Results The expression of LC3 II/I and Atg5 in the vitiligo melanocytes were significantly higher than that in normal controls melanocytes(t=10.36,13.01,all P<0.01),while the expression levels of P62,m TOR protein and tyrosinase activity in vitiligo melanocytes were all significantly lower than those of normal melanocytes(t=5.12,7.22,36.51,all P<0.01).Conclusion The autophagy is upregulated in the vitiligo melanocytes with the decline of melanocyte function,suggesting that autophagy might be involved in the pathogenesis of vitiligo.

关 键 词：白癜风 黑素细胞 自噬 酪氨酸酶活性 

分 类 号：R75[医药卫生—皮肤病学与性病学]