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Photoprotection of HaCaT cells against UVA radiation by melanin-binding hydroxychloroquine in vitro
Author(s): 
Pages: 76-79
Year: Issue:  2
Journal: JOURNAL OF CLINICAL DERMATOLOGY

Keyword:  黑素羟氯喹长波紫外线凋亡HaCaT细胞;
Abstract: 目的:观察与黑素结合的羟氯喹(HCQ)对细胞内活性氧基(ROS)的清除以及对长波紫外线(UVA)诱导人角质形成细胞(HaCaT细胞)凋亡和坏死的保护.方法:自一株产黑素的嗜麦芽假单胞菌AT18的培养液中分离纯化获得细菌衍生黑素,在体外与不同浓度HCQ结合后处理培养的HaCaT细胞,随后给予半数致死剂量(30 J/cm2)的UVA照射.照射12 h后,以四甲基偶氮唑蓝(MTT)比色法测定细胞存活率;碘化丙啶(PI)染色结合流式细胞仪检测细胞凋亡率;采用二氯荧光素二酯(DCFH-DA)标记法测定细胞内ROS水平.结果:低浓度(10、50 μmol/L)HCQ与黑素结合后能明显保护HaCaT细胞抵抗半数致死剂量(30 J/cm2)UVA照射,尤其是50 μmol/L HCQ+50 mg/L黑素处理组细胞存活率较单纯黑素和单纯HCQ组显著增高(P<0.05);相反,用高浓度(250 μmol/L)HCQ与黑素结合处理细胞,细胞存活率较单纯黑素和单纯HCQ组减低.与单纯黑素和单纯HCQ组相比,10 μmol/L HCQ与黑素结合还能显著提高细胞内ROS的清除能力和抑制UVA'诱导HaCaT细胞凋亡.结论:低浓度HCQ与黑素结合能协同增强黑素对细胞内ROS的清除,抑制UVA诱导的皮肤细胞凋亡,这些作用很可能关系到治疗皮肤型红斑狼疮时抗疟药的抗光敏机制.
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