皮肤中原代黑色素细胞和角质形成细胞同时分离法及其在超细颗粒物毒性效应评价中的应用  

Isolation of Primary Human Foreskin Melanocytes/Keratinocytes for Ultrafine Particles’Toxicity Evaluations

作　　者：程战文 殷诺雅 郑卫 Francesco Faiola Cheng Zhanwen;Yin Nuoya;Zheng Wei;Francesco Faiola(State Key Laboratory of Environmental Chemistry and Ecotoxicology,Research Center for Eco-Environmental Sciences,Chinese Academy of Sciences,Beijing 100085,China;College of Resources and Environment,University of Chinese Academy of Sciences,Beijing 100049,China;Department of Urology&Nephrology,The Third Affiliated Hospital of Chongqing Medical University,Chongqing 401120,China)

机构地区：[1]中国科学院生态环境研究中心,环境化学与生态毒理学国家重点实验室,北京100085 [2]中国科学院大学资源与环境学院,北京100049 [3]重庆医科大学附属第三医院泌尿外科,重庆401120

出　　处：《生态毒理学报》2020年第6期158-166,共9页Asian Journal of Ecotoxicology

基　　金：国家自然科学基金创新研究群体项目(22021003);中国科学院战略性先导科技专项(B类)(XDB14040300);国家自然科学基金面上项目(21577166);国家自然科学基金青年科学基金资助项目(21707160)。

摘　　要：大气污染及大气中的超细颗粒物,会对人体健康造成一定的危害。皮肤作为人体最大的器官,是保护人体不受外源性物质损害的第一道防线。为探究超细颗粒物对人体皮肤特别是表皮的潜在风险,从人体皮肤中分离得到了原代黑色素细胞和角质形成细胞,使用1~10000μg·L^(-1)商业化超细碳颗粒物模拟大气中的超细颗粒物,探究了其潜在的皮肤毒性。研究结果表明,从皮肤中分离的原代黑色素细胞和角质形成细胞能在体外扩增,具有相应的功能。超细碳颗粒物粒径<100 nm,处于纳米尺度,且72 h急性暴露不会影响黑色素细胞的细胞活性。qRT-PCR结果显示,超细碳颗粒物暴露会上调黑色素细胞功能基因MITF、MITF-M、c-KIT、SILV、PAX3、SLUG、TYR和TYRP1的表达,干扰其基因表达。以上研究结果为大气颗粒物的毒性评价,提供了重要的数据和模型。Air pollution and ambient ultrafine particulate matter have been proved to affect human health.As the largest organ of the human body,the skin is the first line of defense to protect the body from exogenous substances.To evaluate the potential toxic effects of ultrafine particulate matter on human skin,especially on the epidermis,we simultaneously isolated primary keratinocytes and melanocytes from foreskin at first.Those cells could proliferate in vitro,and maintain their functions.Then,we treated the primary melanocytes with 1~10000μg·L^(-1)commercial ultrafine carbon particles,less than 100 nm in diameter,which mimic ambient ultrafine particulate matter.The ultrafine carbon particles had no effects on cell viability after 72 h exposure.However,qRT-PCR assays revealed that ultrafine carbon could promote the expression of the melanocyte functional genes MITF,MITF-M,c-KIT,SILV,PAX3,SLUG,TYR and TYRP1.These results provide important models and data to assess the toxicity of atmospheric ultrafine particulate matter.

关 键 词：超细碳颗粒物 超细颗粒物 黑色素细胞 角质形成细胞 急性毒性 功能基因 表皮 

分 类 号：X171.5[环境科学与工程—环境科学]