Nie H Q, Wang P, Zhang X Y, Ding C, Liu J, Xu A E
Department of Dermatology, Hangzhou Third Hospital, Affiliated Hangzhou Clinical College, Anhui Medical University , Hangzhou 310009, China.
Zhonghua Yi Xue Za Zhi. 2016 Jul 12;96(26):2064-9. doi: 10.3760/cma.j.issn.0376-2491.2016.26.006.
To evaluate autophagy of melanocytes and its mechanism in patients with vitiligo and to analyze its correlation with clinical types of vitiligo.
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 confocal microscope (LSCFM). Protein expressions of microtube-associated protein light chain 3 (LC3Ⅱ) and microphthalmia-associated transcription factor (MITF), tyrosinase (TYR), tyrosinase related protein (TYRP)1, and TYRP2 were detected by Western blot.
(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 LC3Ⅱ 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).
Autophagy of melanocytes may be present in vitiligo and affect the expression of functional molecules, and is related with clinical type of vitiligo.
评估白癜风患者黑素细胞的自噬及其机制，并分析其与白癜风临床类型的相关性。
2014年5月至2015年6月期间，在杭州市第三人民医院招募了9例节段型白癜风（SV）患者和11例泛发型白癜风（GV）患者。招募6名皮肤健康者作为对照。从SV和GV患者白斑区域周围的正常肤色皮肤以及对照者的包皮中获取表皮黑素细胞进行体外培养。每组培养物包含阴性对照和雷帕霉素（30 nmol/L）亚组。使用透射电子显微镜（TEM）和免疫荧光激光扫描共聚焦显微镜（LSCFM）观察自噬情况。通过蛋白质印迹法检测微管相关蛋白轻链3（LC3Ⅱ）、小眼相关转录因子（MITF）、酪氨酸酶（TYR）、酪氨酸酶相关蛋白（TYRP）1和TYRP2的蛋白表达。
（1）在雷帕霉素处理前，TEM和LSCFM观察到SV组和对照组黑素细胞存在自噬，但GV患者中未观察到。自噬诱导后，SV组、GV组和对照组黑素细胞中自噬体和LC3Ⅱ蛋白表达增加（SV组0.58±0.10对0.37±0.06；GV组0.57±0.16对0.22±0.08；对照组0.67±0.09对0.46±0.12），且GV组自噬强度高于SV组和对照组。（2）自噬诱导前，GV组和SV组患者中MITF、TYR、TYRP1和TYRP2蛋白表达低于对照组；自噬诱导后，三组黑素细胞中MITF、TYR和TYRP1表达均有统计学显著增加（均P<0.05），而TYRP2蛋白表达无显著变化（均P>0.05）。
白癜风患者黑素细胞可能存在自噬并影响功能分子表达，且与白癜风临床类型相关。
