N-乙酰半胱氨酸对化学性缺氧引起HaCaT细胞损伤的保护作用  

Protective effect of N-acetyl cysteine against chemical hypoxia-induced injury to an immortal human skin keratinocyte line HaCaT

作　　者：张美芬[1] 杨春涛[2] 杨战利[2] 孟金兰[3] 曾凡钦[4] 韩艳芳[4] 陈培熹[2] 冯鉴强[2] 

机构地区：[1]中山大学护理学院,广州510080 [2]中山大学中山医学院生理学教研室 [3]广东药学院生理学教研室 [4]中山大学附属第二医院皮肤科

出　　处：《中华皮肤科杂志》2010年第12期859-862,共4页Chinese Journal of Dermatology

基　　金：广东省科技计划（2010B08071035）

摘　　要：目的 探讨N-乙酰半胱氨酸（NAC）能否保护人皮肤永生化角质形成细胞（HaCaT）对抗化学性低氧模拟剂氯化钴（CoCl2）诱导的损伤及对炎症因子的影响.方法 用2000 μmol/L CoCl2处理HaCaT细胞,建立化学性低氧诱导皮肤细胞损伤的实验模型.应用CCK-8比色法检测细胞存活率;ELISA试剂盒检测细胞培养基中IL-6、IL-8和TNF-α的水平;罗丹明123（Rh123）染色/荣光显微镜照相术检测线粒体膜电位（MMP）;谷胱甘肽试剂盒检测细胞内还原型谷胱甘肽（GSH）的含量.结果 不同浓度的NAC预处理HaCaT细胞2 h能明显对抗CoCl2引起的存活率降低;CoCl2处理HaCaT细胞4 h能使IL-6、IL-8和TNF-α的释放显著增加,GSH水平及MMP降低;2000μmol/L NAC预处理2 h能使IL-6和IL-8的释放显著减少,并能使细胞内GSH含量增多,MMP升高.结论 氧自由基清除剂NAC能对抗CoCl2诱导的HaCaT细胞损伤及炎症反应,此作用可能与其减轻细胞内的氧化应激反应有关.Objective To estimate the influences of N-acetyl cysteine （NAC） on a chemical hypoxiamimetic agent CoCl2 induced-injury to, and expressions of inflammatory factors by, an immortal human skin keratinocyte line HaCaT. Methods HaCaT cells were treated with CoCl2 of 2000 μmol/L for 4 hours to set up a chemical hypoxia-induced cell model of skin injury. NAC of various concentrations （ 1000, 2000, 3000 μmol/L）was used to pretreat HaCaT cells for 2 hours prior to the establishment of cell model. After these treatments,cell viability was detected by cell counting kit 8 （CCK-8）, the levels of interleukin 6 and 8 （IL-6 and -8） and tumor necrosis factor α （TNF-α） in culture supernatant by ELISA kits, mitochondrial membrane potential （MMP） by rhodamine 123 （Rh123） staining and photofluorography, intracellular reduced glutathione （GSH）content by glutathione detection kit. Results An obvious decline was observed in HaCaT cell viability after pretreatment with various concentrations of NAC for 2 hours. The treatment with CoCl2 of 2000 μmol/L for 4 hours induced an elevation in the supernatant levels of IL-6, IL-8 and TNF-α and a decrease in GSH content and MMP, while the pretreatment with NAC for 2 hours retarded the CoCl2-induced increase in IL-6 and IL-8 levels as well as decrease in GSH content and MMP. Conclusion The reactive oxygen species （ROS） scavenger NAC can protect against CoCl2-induced injury to and inflammatory reaction in HaCaT cells, which may be associated with a decrement in oxidative stress.

关 键 词：自由基清除剂 缺氧 角蛋白细胞 炎症 

分 类 号：R751[医药卫生—皮肤病学与性病学]