文章摘要
王英英,李凯彬,刘春,王庆,曾伟伟,石存斌,吴淑勤.剑尾鱼脑细胞系的建立及细胞色素P4501A的诱导表达[J].水产学报,2013,37(3):337~343
剑尾鱼脑细胞系的建立及细胞色素P4501A的诱导表达
Development of a brain cell line from swordtail fish, Xiphophorus helleri, and the inducible expression for cytochrome P450 1A
投稿时间:2012-10-30  修订日期:2012-12-23
DOI:DOI:10.3724/SP.J.1231.2013.38402
中文关键词: 剑尾鱼  脑细胞系  细胞色素P4501A
英文关键词: Xiphophorus helleri, brain cell line, cytochrome P450 1A
基金项目:国家自然科学基金资助项目(40976072)
作者单位E-mail
王英英 中国水产科学研究院珠江水产研究所农业部渔药创制重点实验室广东省免疫技术重点实验室 wang-dixiaoye2008@163.com 
李凯彬 中国水产科学研究院珠江水产研究所  
刘春 中国水产科学研究院珠江水产研究所  
王庆 中国水产科学研究院珠江水产研究所  
曾伟伟 中国水产科学研究院珠江水产研究所  
石存斌 中国水产科学研究院珠江水产研究所  
吴淑勤 中国水产科学研究院珠江水产研究所 wushuqin001@21cn.com 
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中文摘要:
      剑尾鱼对多种环境污染物敏感,是生态毒理学研究较为理想的实验动物,相关细胞系的构建可为其系统应用提供基础。为了构建剑尾鱼脑细胞系及探讨其细胞中CYP1A的诱导效应,本研究通过胰蛋白酶消化法对剑尾鱼脑组织进行体外培养,经连续继代培养,建立了可稳定传代的脑细胞系,命名为SFB。SFB最适培养液为含有15% 胎牛血清( FBS) 的DMEM/ F-12和L-15等比混合培养液,培养条件为27℃,5%CO2。生长特性研究表明,第65 代细胞的群体倍增时间为43. 0h,显示出旺盛的生长和分裂能力。染色体分析发现培养细胞的染色体众数为48条,SFB核型公式为2n=2st 46t,臂指数( NF)= 48,与剑尾鱼一致。诱导实验表明:SFB在10-8~10-5mol/L的苯并芘B(a)P诱导下CYP1A mRNA表达量显著提升,且表现出良好的剂量效应关系。因而,脑细胞系的建立为剑尾鱼的毒理学评价研究提供了便利,也为其系统的生态毒理学应用打下基础。
英文摘要:
      Swordtail fish, Xiphophorus helleri, is an ideal experimental animal for Ecological toxicology studies, which is sensitive to environmental contaminants. The construction of related cell lines can provide foundation for its application. The aim of this study were to establish the swordtail fish brain cell line SFB (swordtail fish brain cell line) and to study its CYP1A mRNA inductive effect. Primary brain cell culture of Swordtail fish was initiated by trypsin digestion methods, and a stable brain cell line was obtained successfully after culture and proliferation. The optimal culture medium for SFB was Leibovitz-15 to blend DMEM/F12 medium with 1:1 supplemented with 15% fetal bovine serum, and the optimal culture conditions for SFB were 27℃ and 5% CO2. The SFB grew actively under the optimal medium and culture conditions, the doubling time is 43. 048 hours at 65th passage . And chromosome analysis showed that the SFB exhibited chromosomal a diploidy with a modal chromosome number of 48, the karyotype formula was 2n=2st 46t, NF=48, which consistent with the swordtail fish.The cells maintained their original shape and high viability after cryopreserved and resuscitated. The induction experimental results showed that, the expression of CYP1A mRNA in B(a)P-treated (10-8~10-5mol/L) SFB cells increased significantly, and increased along with the increasing dosage of B(a)P. A dose-dependent relationship between the levels of CYP1A mRNA expression and doses of B(a)P was observed. Thus, the establishment of this cell line will provide convenience for the toxicological evaluation and a basis for its application in Ecological toxicology.
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