大鼠体内碱性彗星试验中阳性药环磷酰胺和甲基磺酸乙酯给药周期的筛选研究
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篇名: 大鼠体内碱性彗星试验中阳性药环磷酰胺和甲基磺酸乙酯给药周期的筛选研究
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摘要: 目的:筛选大鼠体内碱性彗星试验中阳性药环磷酰胺(CP)和甲基磺酸乙酯(EMS)的给药周期。方法:将SD大鼠随机分为空白对照组、CP组(40 mg/kg,腹腔注射)和EMS组(100 mg/kg,灌胃),每组9只,每24 h给药1次,连续给药4次。分别于第2、3、4次给药后3 h(即首次给药后27、51、75 h)各组随机选取3只大鼠,处死并收集肝和胃样本,经过制备单细胞悬液、制片、裂解、解链、电泳及染色后,使用Comet Assay Ⅳ彗星试验数据分析系统进行分析,以彗星尾部DNA百分比(Tail DNA%)、尾长(TL)和尾矩(TM)为评价指标,筛选最佳给药周期。结果:空白对照组大鼠3个检测时间点肝和胃的Tail DNA%、TL、TM差异均无统计学意义(P>0.05),表明彗星试验体系比较稳定。与空白对照组比较,CP组和EMS组大鼠3个检测时间点肝和胃的Tail DNA%、TL(27 h的胃除外)、TM均明显升高(P<0.05)。其中,CP组大鼠3个检测时间点间肝的Tail DNA%、TL、TM差异均无统计学意义(P>0.05),胃的Tail DNA%、TL均在给药51 h时最大,TM随给药时间的延长而升高;EMS组大鼠肝的Tail DNA%、胃和肝的TM均随给药时间的延长而升高。EMS组大鼠胃的Tail DNA%、肝和胃的TL均在给药51 h时最大。结论:在进行体内碱性彗星试验时,建议CP和EMS的给药周期是每24 h给药1次,连续给药3次。
ABSTRACT: OBJECTIVE: To screen the dosing period of positive drug cyclophosphamide (CP) and ethyl methanesulfonate (EMS) in alkaline comet assay of rats in vivo. METHODS: SD rats were randomly divided into blank control group, CP group (40 mg/kg, i.p.) and EMS group (100 mg/kg, i.g.), 9 rats in each group, every 24 h, for consecutive 4 times. 3 h after secondary, third and forth medication (27, 51, 75 h after first medication), 3 rats were randomly selected and sacrificed in each group. Liver and stomach samples were collected. After single cell suspension preparation, production, lysis, chain breaking, electrophoresis and dyeing, the evaluation indicators, including tail DNA%, tail length (TL) and tail moment (TM), Comet Assay Ⅳ comet assay data analysis system was used to screen the optimal dosing period. RESULTS: There was no statistical significance in Tail DNA%, TL and TM of liver and stomach in blank control group at 3 test time points (P>0.05), indicating that the comet assay system was stable. Tail DNA%, TL (except for stomach at 27 h) and TM of liver and stomach in CP and EMS groups were increased significantly, compared with blank control group (P<0.05). There was no statistical significance in Tail DNA%, TL and TM of liver in CP group at the three test time points (P>0.05), tail DNA% and TL of stomach in CP group were maximal at 51 h. TM was increased with dosing period. Tail DNA% of liver in EMS group, TM of stomach and liver were increased with dosing period. Tail DNA% of the stomach, TL of liver and stomach were the highest at 51 h in EMS group. CONCLUSIONS: During in vivo alkaline comet assay, it is recommended that CP and EMS be administered once every 24 h and three times continuously.
期刊: 2018年第29卷第23期
作者: 李丽丽,杨林,谢锋,辛艳飞,张升,陈娇婷,宣尧仙,由振强
AUTHORS: LI Lili,YANG Lin,XIE Feng,XIN Yanfei,ZHANG Sheng,CHEN Jiaoting,XUAN Yaoxian,YOU Zhenqiang
关键字: 体内碱性彗星试验;环磷酰胺;甲基磺酸乙酯;给药周期;大鼠
KEYWORDS: Alkaline comet assay in vivo; Cyclophosphamide; Ethyl methanesulfonate; Dosing period; Rats
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