生态毒理
马陶武,王子健,易浪波.稀有鮈鲫和日本青鳉肝细胞原代培养及其对2,3,7,8-TCDD的敏感性比较[J].环境科学学报,2010,30(6):1243-1249
稀有鮈鲫和日本青鳉肝细胞原代培养及其对2,3,7,8-TCDD的敏感性比较
- Primary culture of hepatocytes from rare minnow (Gobiocypris rarus) and Japanese medaka (Oryzias latipes) and comparison of their sensitivity to 2,3,7,8-TCDD
- 基金项目:国家重点基础研究发展计划(973)项目(No.2007CB407304);湖南省自然科学基金项目(No.05JJ40031);吉首大学科研基金项目
- 马陶武
- 吉首大学生物资源与环境科学学院, 吉首 416000
- 王子健
- 中国科学院生态环境研究中心环境水质学国家重点实验室, 北京 100085
- 易浪波
- 吉首大学生物资源与环境科学学院, 吉首 416000
- 摘要:利用胰酶消化和机械分散相结合的方法对稀有鮈鲫和日本青鳉的肝细胞进行分离和培养,测定了不同培养时间下两种原代培养肝细胞的活力,研究了两种原代培养肝细胞受到2,3,7,8-TCDD暴露后EROD活力变化的时间-和浓度-效应关系.结果表明,稀有鮈鲫和日本青鳉的肝细胞产率分别达到2.0×107cells·g-1和1.5×107cells·g-1,稀有鮈鲫原代肝细胞的活力可以稳定地保持5d,日本青鳉原代肝细胞的活力至少可以稳定地保持1周;不同浓度TCDD暴露后,两种鱼的原代肝细胞EROD活力变化均显示良好的时间-浓度-效应关系,TCDD诱导稀有鮈鲫和日本青鳉原代肝细胞EROD活力的最低可观察效应浓度(LOEC)分别为4pg·mL-1和1pg·mL-1;TCDD诱导日本青鳉原代肝细胞EROD活力的EC50值低于稀有鮈鲫.就原代肝细胞培养操作和对毒物胁迫的敏感性而言,日本青鳉优于稀有鮈鲫.
- Abstract:The hepatocytes from rare minnow(Gobiocypris rarus)and Japanese medaka(Oryzias latipes)were cultured after isolation by digestion with pancreatin along with mechanical separation.The viability of the hepatocytes was determined after primary culture for various periods of time.The time-and concentration-dependent ERODresponses of the primary hepatocytes to 2,3,7,8-TCDDwere measured.The results showed that the hepatocyte yields were 2.0×107 and 1.5×107 cells g·-1 liver tissues for rare minnow and Japanese medaka,respectively.The primary hepatocytes of rare minnow were viable for 5 d whereas those of Japanese medaka were viable for more than one week.Both primary hepatocytes showed good time-and concentration-dependent ERODresponses to 2,3,7,8-TCDD.The lowest concentrations of 2,3,7,8-TCDDinducing the ERODactivity in the primary hepatocytes of rare minnow and Japanese medaka were 4 and 1 pg·mL-1,respectively.The EC50 of 2,3,7,8-TCDDinducing the ERODactivity in the primary hepatocytes of Japanese medaka was lower than that in rare minnow.In terms of the preparation of primary hepatocytes and the sensitivity to the stress toxicants,Japanese medaka was superior to rare minnow.
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