四氢嘧啶对HaCaT细胞活力及AQP3、Claudin-1、ZO-1表达的影响  

Effects of Ectoine on HaCaT Cell Viability and AQP3,Claudin-1 and ZO-1 Expression

作　　者：高翔[1] 严雅军 徐烁 乔丽娟 李欣冉 朱德锐[1] GAO Xiang;YAN Yajun;XU Shuo;QIAO Lijuan;LI Xinran;ZHU Derui(Research Center of Basic Medical Sciences,Medical College of Qinghai University,Xining,China,810016)

机构地区：[1]青海大学医学部基础医学研究中心,西宁810016

出　　处：《中国高原医学与生物学杂志》2024年第2期131-137,共7页Journal of Chinese High Altitude Medicine & Biology

基　　金：青海省基础应用研究计划项目(2022-ZJ-771);青海省2022年“昆仑英才·高端创新创业人才”拔尖人才培养项目;青海大学中青年项目(2020-QNY-5)

摘　　要：目的研究四氢嘧啶(Ectoine)对人角质形成细胞(HaCaT)的细胞活力、细胞凋亡率和细胞活性氧(ROS)的影响,并分析水通道蛋白3(AQP3)、紧密连接蛋白-1(Claudin-1)、闭锁连接蛋白(ZO-1)的表达水平。方法设置Ectoine浓度实验组和空白对照组,培养HaCaT细胞(24 h),采用CCK8法检测HaCaT细胞的细胞活力,甄选最佳的Ectoine作用浓度。用Western Blot法检测HaCaT细胞的AQP3、Claudin-1、ZO-1表达水平;用流式细胞仪检测HaCaT细胞的凋亡率及胞内ROS水平。结果CCK8法检测结果显示,0.10%Ectoine组、0.20%Ectoine组、0.30%Ectoine组的HaCaT细胞活力均高于120%,其中0.20%Ectoine组最高(146.92%±7.67%)。Ectoine浓度实验组相对于空白对照组,HaCaT细胞的AQP3、Claudin-1、ZO-1表达水平明显升高(P<0.05),且细胞凋亡率和胞内ROS水平均明显降低(P<0.05)。结论Ectoine可提高HaCaT细胞的细胞活力,降低凋亡率和胞内ROS水平,上调AQP3、Claudin-1、ZO-1表达水平。Ectoine可能与相关保湿蛋白的表达相关,对HaCaT细胞具有一定的保护作用。Objective The study investigates the effects of Ectoine on cell viability,apoptosis,and intracellular reactive oxygen species(ROS)levels in human keratinocyte cells(HaCaT),as well as analyzing the expression levels of aquaporin 3(AQP3), claudin-1, and zonula occludens-1(ZO-1). Methods Experimental groups with varying concentrations of Ectoine and a blank control group were set up.HaCaT cells were cultured for 24 hours,and the cell viability was detected using the CCK8 method to determine the optimal concentration of Ectoine.The expression levels of AQP3,Claudin-1,ZO-1 in HaCaT cells were examined by Western Blot analysis,while the apoptosis rate and intracellular ROS levels in HaCaT cells were detected using flow cytometry.Results The CCK8results showed that the cell viability in the 0.1%,0.2%,0.3% Ectoine groups were all higher than 120%,with the0.2% Ectoine group demonstrating the highest level at 146.92%±7.67%.Compared with the blank control group,the experimental groups with different concentrations of Ectoine significantly increased the expression levels of AQP3,Claudin-1,and ZO-1 in HaCaT cells(P<0.05),and concurrently decreased both the apoptosis rate and intracellular ROS levels significantly(P<0.05).Conclusion Ectoine can enhance the cell viability of HaCaT cells,decrease their apoptosis rate and intracellular ROS levels,and upregulate the expression of AQP3,Claudin-1,and ZO-1.Ectoine may be related to the expression of relevant moisturizing proteins and exhibits a certain protective effect on HaCaT cells.

关 键 词：四氢嘧啶 人角质形成细胞 水通道蛋白3 紧密连接蛋白-1 闭锁连接蛋白 

分 类 号：R751[医药卫生—皮肤病学与性病学]