Wnt3a减轻黑素细胞iMC65氧化应激损伤  

Wnt3a protects melanocytes iMC65 against oxidative stress

作　　者：郑妍[1] 杨珂[1] 李娅莎[1] 董世访[1] 周丽娜[1] ZHENG Yan YANG Ke LI Ya-sha DONG Shi-fang ZHOU Li-na(Pediatric Research Institute, Children＇s Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation Base of Child Development and Critical Disorders Chongqing Engineering Research Center of Stem Cell Therapy, Chongqing 400014, China)

机构地区：[1]重庆医科大学附属儿童医院儿科研究所儿童发育疾病研究教育部重点实验室儿童发育重大疾病国家国际科技合作基地重庆市干细胞治疗工程技术研究中心,重庆400014

出　　处：《基础医学与临床》2017年第10期1389-1394,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(81371718;81101205)

摘　　要：目的探讨Wnt3a对氧化应激损伤的i MC65黑素细胞的保护作用及机制。方法将黑素细胞分成对照组,Wnt3a组,H2O2处理组,Wnt3a干预组,750μmol/L的H2O2作用模拟黑素细胞的氧化应激损伤。MTT实验检测细胞活性,流式细胞术检测细胞凋亡率和细胞产生活性氧(ROS),荧光素酶报告基因检测Nrf2/ARE通路的激活,Western blot检测Nrf2/ARE通路的相关蛋白表达。结果与对照组相比,H2O2处理组的细胞活性明显下降(P<0.01),凋亡比率明显上升(P<0.01),ROS的产生明显增加(P<0.01)。而Wnt3a干预组能显著缓解H2O2处理组细胞活性的降低(P<0.05)、降低凋亡比率(P<0.05),减少ROS的产生(P<0.05)。Wnt3a也能上调Nrf2和HO-1蛋白水平的表达。结论 Wnt3a可以保护氧化应激状态下的黑素细胞,其机制可能与激活Nrf2/ARE有关。Objective To investigate the protective effect of Wnt3 a on the oxidative stress damage of melanocytes and its mechanism. Methods The melanocytes were divided into 4 groups： control group,Wnt3 a group,H2O2 treatment group,Wnt3 a treatment group. The melanocytes were treated with 750 μmol/L H2O2 for oxidative stress damage. The activity of cells was detected by MTT assay. The apoptosis rate was examined by flow cytometry. The ROS production was observed by fluorescence microscopy and flow cytometry. The Nrf2/ARE pathway activation was determined with an ARE-driven luciferase reporter construct. The expressions of Nrf2 and HO-1 were examined by Western blot. Results Compared with control group,cell activity decreased（ P〈0. 01）,the ratio of apoptosis increased（ P〈0. 01）,ROS production was raised in H2O2 treatment group（ P〈0. 01）. While compared with H2O2 treatment group,cell activity was relieved（ P〈0. 05）,the ratio of apoptosis decreased（ P 0. 05）,ROS production was declined in Wnt3 a treatment group（ P 0. 05）. Wnt3 a up-regulated the transcriptional activity of Nrf2 as determined with improving the relative activity of luciferase regulated by the antioxidant response element ARE. Wnt3 a alsoincreases the expression of Nrf2 and HO-1 protein levels. Conclusions Wnt3 a protects the melanocytes against oxidative injury by H2O2. The mechanism would be related to the activation of Nrf2/ARE pathway.

关 键 词：WNT3A 黑素细胞 NRF2 氧化应激 

分 类 号：R758.41[医药卫生—皮肤病学与性病学]