摘要
目的:探讨黄芩苷对H_2O_2氧化损伤正常人黑素细胞的保护作用及其机制研究。方法:构建H_2O_2氧化损伤黑素细胞模型,通过观察不同浓度的黄芩苷处理H_2O_2氧化损伤正常人黑素细胞,采用倒置显微镜观察黄芩苷对黑素细胞形态的影响;cell counting kit-8(CCK-8)法检测黄芩苷对黑素细胞增殖活力作用;乳酸脱氢酶(lactate dehydrogenase,LDH)法检测黄芩苷对黑素细胞LDH表达的影响,并通过流式细胞仪检测活性氧(ROS)水平,荧光实时定量RT-PCR检测过氧化氢酶(catalase,CAT)、谷胱甘肽过氧化物酶1(peroxidase 1,GPX1)m RNA表达情况,通过ELISA试剂盒检测黄芩苷和H_2O_2处理黑素细胞后分泌的热休克蛋白70(heat shock protein 70,HSP70)。结果:镜下观察黄芩苷给药后细胞较对照组形态明显改变,细胞数增多;黄芩苷作用黑素细胞后增殖率较对照组升高(P<0.05);黄芩苷明显抑制黑素细胞中LDH表达,呈剂量-效应关系(P<0.05)。黄芩苷能显著促进CAT和GPX1m RNA表达,且明显抑制HSP70分泌。结论:黄芩苷对H_2O_2氧化损伤的正常人黑素细胞具有明显保护作用,可为临床白癜风的治疗提供参考。
AIM:To study the mechanism of anti-oxidant and melanocyte protection by Baicalin.METHODS:Different concentration of Baicalin and H2O2 were coeuhured with normal human melano-cyte.Cell proliferation was detected by cell counting kit-8 assay, morphology of the human melanocyte was observed by ordinary inverted microscope,and LDH assay was used to detect cell viability.For mechanism research,DCFH-DA ROS level was assayed by flow cytometry.Induced HSP70 from melanocyte was detected using HSP70 high sensitivity ELISA kit.By RT-PCR,mRNA levels of CAT and GPX1 were dectected.RESULTS:Baicalin has obvious protective effects on melanocytes damage caused by H2O2.Compared with control group,in Baicalin groups,cell shape was changed and increased.In 12.5, 25, 50μmol/L Baicalin groups, the effects of on melanocyte proliferation rate were higher than the control group (P 〈0.05 ).Baicalin inhibited the expression of LDH in black pigment ceils in a dose-dependent manner,there was statistically significant difference(p〈0.05).After H2O2 oxidative damage of melanoeytes,Baicalin could sig-nificantly promote the expression of CAT and GPX1 mRNA,and the secretion of HSP70 was significantly increased.CONCLUSION:Baicalin protects human melanocytes from oxidative damage by inhibiting H2O2induced apoptosis.It is referential for the treatment of vitiligo.
作者
祝逸平
金嵘
林福全
周妙妮
王遂泉
许爱娥
ZHU Yiping;JIN Rong;LIN Fuquan;ZHOU Miaoni;WANG Suiquan;XU Aie(The Third People's Hospital in Hangzhou Affiliated to Zhejiang Chinese Medicine University, Hangzhou 310009, Zhejiang, China)
出处
《中国临床药理学与治疗学》
CAS
CSCD
2018年第4期377-382,共6页
Chinese Journal of Clinical Pharmacology and Therapeutics
基金
浙江省自然科学基金(LQ16H290001
LY18H110001)
国家自然科学基金(81472887
81773335)
浙江省基础公益研究计划项目(LGF18H110002)
国家临床重点专科建设项目