亚砷酸钠对全反式维甲酸诱导的小鼠神经干细胞活力及分化的影响  

Effects of sodium arsenite on cell viability and differentiation of mouse neural stem cells induced by ATRA

作　　者：艾合买提·艾不拉 白娇娇 孙红光 申晨晨 古力乃则尔·阿不都外力 马艳[1] Aihemaiti Aibula;BAI Jiaojiao;SUN Hongguang;SHEN Chenchen;Gulinaizeer Abuduwaili;MA Yan(School of Public Health,Xinjiang Medical University,Urumqi 830011,China)

机构地区：[1]新疆医科大学公共卫生学院,乌鲁木齐830011

出　　处：《山东医药》2023年第20期34-38,共5页Shandong Medical Journal

基　　金：国家自然科学基金项目(81760568)。

摘　　要：目的 探究亚砷酸钠(NaAsO_(2))对全反式维甲酸(ATRA)诱导的小鼠神经干细胞NE-4C细胞活力及分化的影响。方法 分别以0、0.5、1.0、5.0μmol/L的ATRA诱导NE-4C细胞,采用CCK-8法检测细胞活力,采用实时荧光定量PCR法检测NE-4C细胞中的神经干细胞特异性标志物[巢蛋白(Nestin)、性别决定区Y框蛋白2(Sox2)]、神经元特异性标志物[神经元特异性烯醇化酶(NSE)、RNA结合蛋白fox1(Rbfox1)、III类β-微管蛋白(βⅢ-Tubulin)]、星形胶质细胞特异性标志物胶质纤维酸性蛋白(GFAP)、少突胶质细胞标志物髓鞘碱性蛋白的mRNA,筛选ATRA的适宜作用浓度。将适宜浓度的ATRA作用于NE-4C细胞,建立神经干细胞分化细胞模型,分为对照组和实验1、2、3、4组,实验1、2、3、4组分别给予0.3、0.6、0.9、1.2μmol/L的NaAsO_(2)干预,对照组不给予药物;培养7 d后,采用CCK-8法检测细胞活力,免疫荧光染色观察细胞分化情况,采用流式细胞术检测细胞中的Nestin、NSE、βⅢ-Tubulin、GFAP。结果 ATRA作用后,NE-4C细胞活力降低,Sox2、Nestin mRNA表达下调,βⅢ-Tubulin、NSE、Rbfox1 mRNA表达升高,作用呈浓度依赖性,0.5μmol/L的ATRA作用时细胞活力和分化相关基因表达最高(P均<0.05)。建立神经干细胞分化细胞模型后,对照组和实验1、2、3、4组NE-4C细胞活力和Nestin、NSE、βⅢ-Tubulin、GFAP表达依次降低(P均<0.05)。结论 NaAsO_(2)可降低ATRA诱导的NE-4C细胞活力,并抑制NE-4C细胞向神经元分化,NaAsO_(2)浓度越高则其对NE-4C细胞活力和分化的抑制作用越强。Objective To investigate the effects of sodium arsenite(NaAsO_2) on the cell viability and differentiation of mouse neural stem cells(NE-4C) induced by all-trans retinoic acid(ATRA).Methods NE-4C cells were induced by 0,0.5,1.0 and 5.0 μmol/L ATRA,and the cell viability was detected by CCK-8.Neural stem cells(NSCs) specific markers [Nestin and sex determining region Y-box 2(Sox2)],neuron-specific markers [neuron-specific enolase(NSE),RNA binding protein fox-1 homolog 1(Rbfox1),and βⅢ-Tubulin],glial fibrillary acidic protein(GFAP),and myelin basic protein(MBP) mRNA were detected by real-time quantitative fluorescent PCR(RT-PCR),and the appropriate concentration of ATRA was screened.NE-4C cells were treated with ATRA at appropriate concentration to establish the neural stem cell differentiation models,which were then divided into the control group and experimental groups 1,2,3 and 4.Experimental groups 1,2,3 and 4 were treated with 0.3,0.6,0.9 and 1.2 μmol/L NaAsO_2,respectively,while the control group was not treated with any drug.After 7 days of culture,CCK-8 method was used to detect cell viability,immunofluorescence staining was used to observe cell differentiation,and flow cytometry was used to detect Nestin,NSE,βⅢ-Tubulin and GFAP in cells.Results After ATRA treatment,the cell viability of NE-4C cells decreased,the mRNA expression levels of Sox2 and Nestin decreased,and the mRNA expression levels of βⅢ-tutubulin,NSE and Rbfox1 increased,in a dose-dependent manner;the cell viability and the expression of differentiation-related genes were the highest in the treatment of 0.5 μmol/L ATRA(P<0.05).After the neural stem cell differentiation model was established,the cell viability of NE-4C cells and the expression levels of Nestin,NSE,βⅢ-Tubulin and GFAP in the control group and experimental groups 1,2,3 and 4 decreased successively(all P<0.05).Conclusions NaAsO_(2) can decrease the cell viability of NE-4C cells induced by ATRA and inhibit the differentiation of NE-4C cells into neurons.The higher th

关 键 词：神经干细胞 全反式维甲酸 亚砷酸钠 神经元 细胞分化 

分 类 号：R114[医药卫生—卫生毒理学]