微囊藻毒素-LR对传代细胞的毒性  

Cytotoxicity of microcystin-LR on cultured cells

作　　者：谷康定[1] 林群馨[2] 

机构地区：[1]华中科技大学同济医学院环境医学研究所,武汉430030 [2]香港城市大学生物学与化学系

出　　处：《中国公共卫生》2004年第4期411-412,共2页Chinese Journal of Public Health

基　　金：国家自然科学基金 (2 0 2 770 1 3)

摘　　要：目的　从多种传代细胞株中 ,筛选对微囊藻毒素 -LR敏感的细胞株 ,探讨建立经济、简便研究该毒素传代细胞模型的可能性。方法　不同宿主来源的 8种细胞株 (KB细胞 ,NIH/ 3T3细胞 ,H - 4 -Ⅱ -E细胞 ,Hela细胞 ,Vero细胞 ,HepG2细胞 ,Caco - 2细胞 ,HL - 6 0细胞 )与不同浓度的微囊藻毒素 -LR作用 ,细胞培养 2 4 ,4 8,72 ,96h后观察其形态学变化 ,利用体外细胞法 (MTT)测定其细胞毒性终点 (判断细胞活性 )及乳酸脱氢酶试验测定细胞膜受损情况。结果　 8种细胞株中 ,KB细胞和H - 4 -Ⅱ -E细胞在培养 96h ,毒素浓度大于 18 8μg/ml时 ,呈现明显剂量 -反应关系。KB细胞与毒素接触 8h后就有显著的形态学改变 ,其实验结果的重现性和稳定性非常好 ;乳酸脱氢酶试验显示微囊藻毒素 -LR可导致KB细胞膜损伤。结论　微囊藻毒素 -LR对KB细胞和H - 4 -Ⅱ -E细胞有明显的细胞毒性作用。从形态学变化、试验结果的重现性、稳定性和敏感性考虑 ,KB细胞可用作研究该毒素的传代细胞模型。Objective Cell lines were empolyed to explore the possibility of taking permanent cell lines as models for detecting microcystins and studying their mechanism of cytotoxicity.Methods Eight kinds of cell strains from different hosts(KB,NIH/3T3,H-4-II-E,Hela,Vero,Hep G2,Caco-2,HL-60 cell) were mixed with purified microcystin-LR and incubated for 24,48,72 and 96 hours;Meanwhile morphological alterations,colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays,lactate dehydrogenase (LDH) release assay were respectively used to evaluate the end point of toxicity.Results Among 8 cell lines,KB and H-4-II-E cell lines showed significant dose-response effect at toxin concentrations greater than 18.8μg/ml with 96-hour incubation.In KB cells the toxin induced marked morphological alterations after incubation for 8 hours which was with good reproducibility and consistency.LDH assay results showed that microcystin LR caused cell membrane damage.Conclusion Purified microcystin-LR did induce toxic effect on KB and H-4-II-E cell lines.However,considering morphological changes,reproducibility and consistency,KB cells seemed to be a better potential model for evaluating cytotoxicity of microcystin-LR.

关 键 词：传代细胞 微囊藻毒素-LR 细胞毒性 

分 类 号：R114[医药卫生—卫生毒理学]