白癜风患者皮肤黑素细胞自噬与临床类型的相关性  

Relationship between autophagy of melanocytes in patients with vitiligo and clinical types

作　　者：聂慧琼 王平[1] 张小燕[1] 丁承[1] 刘娇[1] 许爱娥[1] 

机构地区：[1]安徽医科大学杭州临床学院(杭州市第三人民医院)皮肤科,310009

出　　处：《中华医学杂志》2016年第26期2064-2069,共6页National Medical Journal of China

基　　金：国家自然科学基金（81071294）;浙江省医药卫生科技计划项目（2014KYB200）;浙江省公益技术研究社会发展项目（2016C33206）;杭州市医疗卫生及重点专科专病科研攻关专项（20140733Q19）

摘　　要：目的研究白癜风患者皮肤黑素细胞自噬及其机制，分析黑素细胞自噬与白癜风临床类型的相关性。方法 选取2014年5月至2015年6月杭州市第三人民医院就诊的节段型白癜风（SV）患者9例（SV组）、泛发型白癜风（GV）患者11例（GV组），另取6例行包皮环切手术患者的切除包皮作为正常皮肤对照组。取白癜风患者白斑区周围正常肤色部位皮肤和对照组包皮的表皮黑素细胞进行体外培养，采用雷帕霉素30 nmol/L进行自噬诱导。采用透射电子显微镜观察自噬现象，荧光共聚焦显微镜观察自噬流强度，免疫印迹检测自噬标记性蛋白——微管相关蛋白1轻链3Ⅱ型（LC3Ⅱ）和黑素细胞功能性分子——小眼畸形相关转录因子（MITF）、酪氨酸酶（TYR）、酪氨酸酶相关蛋白1和2（TYRP1、TYRP2）蛋白含量表达。结果（1）自噬诱导前，对照组和SV组黑素细胞均观察到存在自噬现象，而GV组黑素细胞未见自噬；自噬诱导后，SV、GV和对照组黑素细胞自噬体数量均增多，LC3Ⅱ蛋白含量增加（SV组：0.58±0.10比0.37±0.06；GV组：0.57±0.16比0.22±0.08；对照组：0.67±0.09比0.46±0.12，均P〈0.01），且GV组自噬强度明显强于SV组和对照组。（2）自噬诱导前，SV和GV组黑素细胞的MITF、TYR、TYRP1和TYRP2蛋白含量低于对照组；自噬诱导后，3组中MITF、TYR和TYRP1蛋白表达量均明显增加（均P〈0.05），而TYRP2表达水平诱导前、后差异无统计学意义（均P〉0.05）。结论白癜风患者黑素细胞存在自噬现象并影响功能性分子的表达，与临床类型相关。Objective To evaluate autophagy of melanocytes and its mechanism in patients with vitiligo and to analyze its correlation with clinical types of vitiligo. Methods Nine cases of segmental vitiligo （SV） and 11 cases of generalized vitiligo （GV） were recruited in Hangzhou Third Hospital between May 2014 and June 2015. Six people with healthy skin were recruited as controls. Epidermal melanocytes were obtained from the normal colour skin around the white spot area in SV and GV patients and from foreskin in controls for culture in vitro. Cultures for each group contained negative control and rapamycin （ 30 nmol/L） sub-groups. The autophagy was observed using transmission electron microscopy （TEM） and immunofluorescence laser scanning eonfocal microscope （LSCFM）. Protein expressions of microtubeassociated protein light chain 3 （ LC3 Ⅱ ） and microphthalmia-associated transcription factor （ MITF ）, tyrosinase （TYR）, tyrosinase related protein （TYRP） 1, and TYRP2 were detected by Western blot. Results （ 1 ） Autophagy of melanocytes was observed under TEM and LSCFM in both SV and control groups before rapamycin treatment, but not in GV patients. The expressions of autophagosome and LC3 Ⅱ protein were increased in melanocytes in SV, GV and control groups after autophagy induction （ SV group 0.58 ± 0.10 vs 0.37 ± 0.06; GV group 0.57 ± 0.16 vs 0.22 ± 0.08 ; control group 0.67 ± 0.09 vs 0.46 ± 0.12）, and the autophagy intensity was higher in the GV group compared to the SV group and the control group. （2） Before autophagy induction, the expressions of MITF, TYR ,TYRP1, and TYRP2 protein were lower in the GV and SV patients compared with the control group; after autophagy induction, the expressions of MITF, TYR, and TYRP1 statistically significantly increased in melanocytes in all the three groups（ all P 〈 0.05 ）, while TYRP2 protein expression was not significantly changed （ all P 〉 0. 05 ）. Conclusions Autophagy of melanocytes may be present in vitiligo and

关 键 词：白癜风 自噬 黑素细胞 

分 类 号：R758.41[医药卫生—皮肤病学与性病学]