过氧化氢诱导人黑素细胞氧化性损伤对TLRs表达的影响  

Effect of Hydrogen Peroxide on the Expression of TLRs Induced by Human Melanocyte Oxidative Damage

作　　者：欧柏生[1] 邓玉霞 赵进 冯杲[1] 黄涛[1] 王艺[1] 张衍[1] 黄玲娟[1] Ou Baisheng;Deng Yuxia;Zhao Jin(First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine,Guangxi 530023,China)

机构地区：[1]广西中医药大学第一附属医院,南宁530023 [2]广西中医药大学,530001 [3]广西壮族自治区皮肤病防治研究所,530003

出　　处：《医学研究杂志》2018年第9期25-28,38,共5页Journal of Medical Research

基　　金：国家自然科学基金资助项目(81460732);广西壮族自治区自然科学基金资助项目(2014GXNSFAA118141)

摘　　要：目的观察过氧化氢(H_2O_2)诱导人黑素A375细胞出现氧化性损伤后,TLRs的表达变化与白癜风病理过程的关系。方法利用不同浓度的过氧化氢处理A375细胞,分为空白对照组(不予H_2O_2处理)及3个不同浓度(100、500和1000μmol/L) H_2O_2处理的实验组,分别作用24、48、72h后,通过黑色素含量检测试剂盒检测诱导后细胞的黑色素含量变化,CCK8检测细胞氧化损伤程度,同时通过q PCR检测过氧化氢诱导后A375细胞中TLRs的表达变化。结果 500μmol/L和1000μmol/L的H_2O_2能显著诱导A375细胞损伤,其中500μmol/L处理72h时细胞活性损伤率约为50%,1000μmol/L过氧化氢处理72h时细胞活性损伤> 50%。黑色素含量检测显示,500μmol/L和1000μmol/L过氧化氢处理后A375细胞中黑色素含量降低最显著。q PCR检测结果显示TLRs家族基因的表达均不同程度上调。结论综合评估过氧化氢诱导后细胞活性和细胞内黑色素含量的变化,确定500μmol/L处理72h为A375细胞氧化应激损伤的最佳诱导条件。初步提示TLRs家族可能在氧化应激诱导的黑色素细胞损伤的过程中发挥重要作用,为开发新的药物靶点提供依据。Objective To observe the relationship between the changes of TLRs and the pathological process of vitiligo after oxidative damage induced by H2O2 in human melanoma A375 cells. Methods A375 cells were treated with different concentrations of H2O2 and divided into blank control group （without H2O2 treatment） and three experimental groups （100μmol/L, 500μmol/L and 1000μmol/L H2O2）. After 24, 48 and 72 hours, the changes of melanin content in cells were respectively detected by melanin detection kit; the degree of oxidative damage was detected by CCK8; the expression of TLRs in A375 cells induced by hydrogen peroxide was detected by qPCR. Results 500μmol/L and 1000μmol/L H2O2 could significantly induce A375 cell′s injury. The cell viability injury rate was about 50% at 500μmol/L H2O2 for 72h and more than 50% at 1000μmol/L H2O2 for 72h. Melanin detection showed the most significant decrease in melanin content in A375 cells after treatment with 500μmol/L and 1000μmol/L H2O2. qPCR test results showed that TLRs family gene expression was significantly up -regulated. Conclusion The comprehensive evaluation of cell viability and melanin content induced by H2O2 was performed to determine the best induction conditions for oxidative stress injury in A375 cells treated with 500μmol/L for 72h. It is suggested that the TLRs family may play an important role in oxidative stress-induced melanocyte injury and provide a basis for the development of new drug targets.

关 键 词：过氧化氢 A375 TLRS 白癜风 

分 类 号：R75[医药卫生—皮肤病学与性病学]