慢性砷暴露对人皮肤角质形成细胞药物转运相关蛋白mRNA表达和砷代谢的影响  

Effects of chronic arsenic exposure on mRNA expression of drug transport proteins and arsenic metabolism in human keratinocytes

作　　者：武瑞瑞[1] 武夏芳 陈佳雯[2] 鞠明光 胡玉鑫[3] 高岚岳[3] 徐苑苑[1,3] 

机构地区：[1]中国医科大学公共卫生学院环境毒理学研究室,辽宁沈阳110122 [2]中国医科大学临床一系 [3]中国医科大学公共卫生与预防医学实验教学示范中心

出　　处：《环境与健康杂志》2016年第9期777-780,共4页Journal of Environment and Health

基　　金：国家自然科学基金青年项目(81302391);辽宁省高等学校攀登学者项目;辽宁省教育厅优秀人才资助项目(LJQ2014090)

摘　　要：目的探讨慢性砷暴露对人皮肤角质形成细胞(human keratinocytes,HaCaT)药物转运蛋白mRNA表达和砷代谢的影响。方法将HaCaT细胞随机分为慢性对照(不予以砷处理)组和慢性砷暴露(100 nmol/L亚砷酸钠染毒28周)组,培养于含10%胎牛血清和1%双抗的培养基中,收集处于对数生长期的细胞,采用实时荧光定量(real-time quantitative PCR)法检测细胞多药耐药相关蛋白mRNA的表达水平。将处于对数生长期的HaCaT对照细胞和慢性砷暴露细胞分别暴露于含终浓度为0(对照)、2.5、5、10、20、30、40、50、60、80、100、150μmol/L亚砷酸钠的培养基中孵育24 h,采用MTS法检测细胞活性。将慢性砷暴露组细胞和对照细胞予以10μmol/L亚砷酸钠处理24 h,采用氢化物发生-原子吸收分光光度法检测细胞砷蓄积和砷排放量。结果与对照细胞相比,慢性砷暴露HaCaT细胞多药耐药相关蛋白ABCC2和ABCC4 mRNA表达水平均升高,差异有统计学意义(P<0.05);而ABCC1、ABCC3、ABCC5、ABCG1、ABCG2 mRNA的表达水平均无显著变化。与对照细胞相比,慢性砷暴露HaCaT经各浓度急性砷处理后细胞存活率均升高,差异有统计学意义(P<0.05)。与对照细胞相比,10μmol/L亚砷酸钠染毒24 h后,慢性砷暴露细胞内的砷蓄积量降低,而砷排放量升高,差异均有统计学意义(P<0.05)。结论慢性砷暴露的HaCaT细胞药物转运蛋白mRNA的表达升高,砷排出能力增强,对砷毒性产生耐受性。Objective To explore the effects of chronicsodium arsenite （NaAsO2） exposure on the mRNA expression of drug transport proteins and arsenic metabolism in human keratinocytes （HaCaT）. Methods The cells were incubated in medium containing 10%FBS and 1%P/S,and were randomly divided into chronic control and NaAsO2 chronic exposure group. The mRNA levels of multidrug resistance proteins were measuredby real-time fluorescence quantitative PCR（RT-PCR）. The HaCaT cells in logarithmic phase were used, treated with NaAsO2 at the doses of 0 （control）, 2.5,5,10,20,30,40,50,60,80,100,150 μmol/L respectively for 24 h. Cell viabilitywas determined by cell proliferation assay （MTS）. Cellular accumulation and efflux of arsenic were detected by hydride generation-atomic absorption spectrophotometry after treatment with 10μmol/L NaAsO2 for 24 h in control and chronic NaAsO2 group. Results Compared with the control group,the mRNA levels of ABCC2 and ABCC4 in chronic NaAsO2 group were higher than control cells （P〈0.05）,while the mRNA levels of ABCC1 ,ABCC3,ABCC5,ABCG1, and ABCG2 did not change significantly. Compared with the control group,the cell viabilities increased significantly in chronic NaAsO2 group after treatment with NaAsO2 for 24 h （P〈0.05）. Compared with the control group, cellular arsenic accumulation decreased in 40 μmol/L NaAsO2 group with 24 h-exposure,while 24 h-arsenic efflux increased （P〈0.05）. Conclusion The mRNA expression of drug transport proteins in HaCaT cells exposed to chronic arsenic is elevated and the arsenic excretion capacity is enhanced, which making HaCaT cells resistant to arsenic toxicity.

关 键 词：砷 皮肤角质形成细胞 多药耐药相关蛋白 砷代谢 

分 类 号：R994.6[医药卫生—毒理学]