二(喹唑啉-4-基)二硒醚体外抗弓形虫速殖子的实验研究

摘要

目的 观察二(喹唑啉-4-基)二硒醚体外对弓形虫RH株速殖子杀伤作用,探讨该药抗弓形虫的效果与机制.方法 将二(喹唑啉-4-基)二硒醚,(下称药物)浓度分为4组,即生理盐水对照组(A组)、药物浓度为0.05 μg/mL(B组)、0.5μg/mL(C组)、5.00 μg/mL(D组),分别加入到含弓形虫速殖子(含虫体4×106个/mL)的24孔培养板中,于1h、2h、4h后收集弓形虫速殖子,以Trypan blue染色法检测弓形虫着色率；C组和A组以MTT法检测弓形虫活性,Giemsa染色法观察虫体形态和结构变化,透射电镜观察弓形虫速殖子超微结构.结果 不同浓度(0.05 μg/mL、0.5 μg/mL、5.00 μg/mL)弓形虫经二(喹唑啉-4-基)二硒醚作用1h后的着色率分别为5.00％、26.70％和99.50％,作用2h后的着色率分别为22.00％、98.70％和100.00％,作用4h后的着色率分别为58.30％、100.00％和100.00％；生理盐水组1h、2h、4h后的着色率分别为1.0％、1.20％和1.70％.0.5 μg/mL浓度药物组,MTT检测弓形虫活性,OD值随时间延长增长率明显低于生理盐水对照组(P＜0.05)；Giemsa染色观察,弓形虫速殖子随着时间的延长,逐渐出现肿胀,两端变圆,虫体坏死现象；透射电镜观察随着时间延长,逐渐出现虫体肿胀,胞膜与基质之间空隙明显,越来越多和大的空泡形成,胞膜出现断裂,内部结构溶解模糊.结论 二(喹唑啉-4-基)二硒醚体外可部分或全部杀死弓形虫,效果与时间、剂量相关.%The effect of 1,2-di (quinazolin-4-yl) diselane on Toxoplasma gondii tachyzoites in vitro was explored in this study.There are four groups:normal saline group (group A),1,2-di (quinazolin-4-yl) diselane 0.05 μg/mL (group B),0.5 μg/mL (group C),and 5 μg/mL (group D).Each group was added into 1 mL T.gondii tachyzoites (4× 106/mL) aqueous suspension.At 1 h,2 h and 4 h after drug treatment,tachyzoites were counted by light microscope with Trypan blue staining.Group C and A was analyzed using MTT and Giemsa staining.The ultrastructure of tachyzoites was observed by transmission electron microscopy.At 1 h treated by 0.05 g/mL,0.5 g/mL,and 5.00 g/mL 1,2-di (quinazolin-4-yl) diselane,5.00％,26.70％ and 99.50％ of T.gondii tachyzoites were stained.And then at 2 h and 4 h,22.00％,98.70％,100.00％and 58.30％,100.00％,100.00％ of T.gondii tachyzoites were stained,respectively.The staining rate of T.gondii tachyzoites of normal saline group was 1.0％,1.20％,and 1.70 ％ at 1 h,2 h and 4 h,respectively.In tachyzoite viability test by MTT,group C was significantly lower than controlgroup A in OD value (P＜0.05).In Giemsa staining test,tachyzoites were swelling,blunt or round in the two ends and underwent necrosis as time prolonging.With the extension of time,it was found that tachyzoites in group C showed swelling and enlarged; apparent gap appeared between cell membrane and matrix; more and bigger vacuoles were formed; the cell membrane became indistinct,broken and damaged.It's suggested that the 1,2-di (quinazolin-4-yl) diselane has a remarkable effect on T.gondii tachyzoites,and the effect of drug has dependency correlation with the concentration and time of action.