人肝微粒-小鼠3T3细胞联合模型评价氟咯草酮代谢后的体外细胞毒性  

Evaluation of fluorochloridone metabolism-dependent cytotoxicity with human liver microsome and mouse 3T3 cell system

作　　者：胡玥[1] 周谡[1] 刘诗宏[2] 石劲敏[1] 周志俊[2] 唐黎明[1] 

机构地区：[1]上海市食品药品检验所药理毒理室/药物安全评价中心,上海201203 [2]复旦大学公共卫生学院

出　　处：《毒理学杂志》2017年第5期335-339,共5页Journal of Toxicology

基　　金：国家自然科学基金(81373040)

摘　　要：目的建立人肝微粒-小鼠3T3细胞联合模型,并应用该模型评价氟咯草酮代谢后的体外细胞毒性。方法以环磷酰胺和他莫西芬分别作为代谢增毒和代谢减毒的模型药物,建立人肝微粒体-小鼠3T3细胞联合模型。首先将环磷酰胺和他莫西芬分别配制成一系列浓度的工作溶液,加入人肝微粒体系孵育180 min后离心,将上清液与培养基1∶1混合获得孵育液。将孵育液与未经代谢的细胞供试液同时给予小鼠3T3细胞,给药48 h后,开展CCK-8试验,利用测得的吸光度(A)值,计算细胞供试液及孵育液的半数抑制浓度IC50以及IC50比值,并以IC50比值为指标评价模型药物的代谢毒性。待测化合物氟咯草酮采用与模型药物相同的方法。结果环磷酰胺IC50比值大于535.31%,经人肝微粒体代谢后对3T3细胞毒性显著增加。他莫西芬IC50比值小于5.58%,经代谢后对3T3细胞毒性显著降低。符合二者作为代谢增毒和代谢减毒代表性药物的特征。氟咯草酮IC50比值大于391.38%,表明经代谢后对3T3细胞毒性增加。结论成功建立人肝微粒-小鼠3T3细胞联合模型。待测化合物氟咯草酮经代谢后毒性增加,为进一步的毒理学研究提供参考依据。Objective The purpose of this study is to evaluate fluorochloridone metabolism-dependent cytotoxicity with human liver microsome and mouse 3T3 system. Methods Cyclophosphamide and Tamoxifen were model toxicants for metabolite toxicity and detoxification, and the human liver microsome and mouse 3T3 cell system was established. The working solution of series of concentrations were added into the HLM incubation system for 180 min, and centrifuged. The supernatant was 1：1 mixed with the culture medium. Then the mouse 3T3 cells were treated for 48 h with the incubation fluid and testing solutions which were not incubated. Using CCK-8 assay to measure the absorbance of each well in the plate. Fitting the curve, and calculating the IC50 values and ratio. IC50 ratio was used to evaluate the metabolite toxicity. Fluorochloridone was tested with the same method as above. Result The IC50 ratio of cyclophosphamide was more than 535.31% ,which indicated that the cytotoxicity was enhanced by the presence of HLM. The IC50 ratio of tamoxifen was less than 5.58% , which demonstrated a lower cytotoxicity in the 3T3 cells after HLM incubation, suggesting hepatic detoxification. The IC50 ratio of fluorochloridone was more than 391.38% , with the enhancing effects through hepatic metabolism. Conclusion The human liver microsome and mouse 3T3 cell system is successfully established. The cytotoxicity of fluorochloridone is enhanced by HLM metabolism, which will support future studies of fluorochloridone induced toxicity research.

关 键 词：人肝微粒体-小鼠3T3细胞联合模型 氟咯草酮 代谢细胞毒性 

分 类 号：R965[医药卫生—药理学] R114[医药卫生—药学]