二(喹唑啉-4-基)二硒醚体外抗弓形虫速殖子的实验研究

Effects of 1 , 2-di(quinazolin-4-yl) diselane inhibited Toxoplasma in vitro

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0.05μg/mL(B组)、0.5μg/mL(C组)、5.00μg/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(0.05μg/mL、0.5μg/mL、5.00μg/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%。0.5μg/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 μg/mL, 0.5 μg/mL, and 5.00 μg/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 control group 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.