白癜风皮肤组织角质形成细胞释放警报素分子的研究

The study of release of alarmin HMGB1 in keratinocytes from skin tissues in vitiligo

目的:探讨人永生化角质形成细胞(HaCaT)和人白癜风组织表达与释放高迁移率蛋白B1(HMGB1)。方法:HaCaT分别经紫外线照射、加凋亡诱导剂、细胞因子刺激、反复冻融处理,细胞培养上清经浓缩后,Western-blot观察24h后HMGB1的释放。免疫荧光染色检测白癜风患者和对照组的活检皮肤组织HMGB1和激活的半胱氨酸蛋白酶3(cleaved caspase-3)的表达。结果:HaCaT经各种处理后,培养上清中均能检测出HMGB1,对照组则不能。白癜风病人角质层检测到cleaved caspase-3,对照组病人未检测到;对照组HMGB1表达在角质形成细胞核内,白癜风组织HMGB1从细胞核转移至细胞浆。结论:HMGB1能够由角质形成细胞在凋亡和坏死时被动释放或者在病理状态下(白癜风)主动分泌。

Objective ：To investigate the release of HMGB1 by HaCaT cell and keratinocytes of tissue from vitiligo patients .Methods ：HaCaT cells were treated with UV light or with apoptosis drugs or with cytokines and freeze-thaw process . The supernatant was condensed and subjected to SDS-PAGE to detect the release of HMGB1 .Measure the HMGB1 and cleaved caspase-3 expression in skin biopsies of normal control subjects and viti-ligo patients by immunofluorescence .Results ：HMGB1 was detected in the supernatant of HaCaT after treated and control supernatant was not detected .Cleaved caspase-3 was found in the stratum corneum of vitiligo patients and control patient was not found .HMGB1 was expressed in the nucleus of keratinocytes of control patients and trans-ferred from nucleus to cytoplasm in vitiligo patients .Conclusion：HMGB1 can be passively released by apoptosis and necrosis keratinocytes or actively released in vitiligo .