中药大蒜素体外抗弓形虫效应及其机制的研究

目的 观察大蒜提取物体外抗弓形虫的作用效果及其机制.方法 将弓形虫RH株速殖子与兔肾细胞共同培养,加入不同浓度的大蒜素(实验组)和磺胺嘧啶(阳性对照组),培养不同时间后取出细胞,固定染色后观察细胞感染率及每个纳虫泡中弓形虫速殖子的数量;采用MTT比色法观察大蒜素对弓形虫速殖子侵袭细胞及其正常细胞增殖的影响;采用台盼兰着色法观察大蒜素对弓形虫速殖子活性的影响;采用TUNEL末端标记法检测弓形虫速殖子凋亡率,对药物的效应和机制进行探讨.结果 (1)10～80 μg/ml的大蒜素能抗弓形虫的感染,呈现剂量依赖性,与时间无明显的相关性.(2)40 μg/ml、80 μg/ml的大蒜素不能抑制细胞的增殖,对细胞无明显的毒副作用;160 μg/ml的大蒜素对细胞有明显的毒副作用.(3)大蒜素80 μg/ml时,台盼兰着色率最高,弓形虫的活力最低.(4)大蒜素80 μg/ml的浓度时,弓形虫速殖子凋亡率最高.结论 大蒜素可以抑制弓形虫速殖子的活力、对宿主细胞的感染力、在细胞内的增殖,无明显的毒副作用,最适宜浓度为80 μg/ml.大蒜素是一种良好的抗弓形虫药物,诱导弓形虫速殖子凋亡是其抗弓形虫机制之一.

Effect and mechanism of Allitridi against Toxoplasma gondii in vitro

Objective To observe the effect and mechanism of allitridi against Toxoplasma gondii in vitro. Methods The Toxoplasma gondii ( RH strain ) and the kidney cells of rabbit were co-cultured, adding different concentration of allitridi and Sulphadiazine. The cultured cells were taken out after 24 h or 48 h, fixed and strained. The infected rate and the number of Toxoplasma gondii per parasitophorous vacuole were observed; The effect of allitridi inhibiting the proliferation of cells invaded by Toxoplasma gondii tachyzoite and normal cells were detected by MTT; The effect of allitridi inhibiting the vigor of Toxoplasma gondii tachyzoite were observed by placenta staining; And the effect of allitridi inducing the apoptosis of Toxoplasma gondii tachyzoite were measured by TUNEL. Results ( 1 ) 10 ～ 80 μg/ml allitridi could resist the infection of toxoplasma gondii in dosage-dependent way, but there wasn't very obvious time-dependent effect; (2) With little side-effect to the cells, 40 μg/ml and 80 μg/ml allitridi couldn't inhibit the proliferation of cells; 160 μg,/ml allitridi had an obvious side-effect to the cells; (3) Under 80 μg/ml allitridi, the rate of Typanblue colouring was the highest, and the vigor of Toxoplasma gondii tachyzoite was the lowest; (4) Under 80 μg/ml allitridi, the rate of apoptosis of Toxoplasma gondii tachyzoite was the highest. Conclusion Allitridi could inhibit the vigor of tachyzoite, restrain its ability to infect the host's cells and its proliferation in the cells while showing no obvious poisonous side effects. The optimal effective concentration was 80 μg/ml. Allitridi is a good medicine against Toxoplasma gondii. One of the mechanisms of its anti-Toxoplas ma gondii is to induce the apoptosis of Toxoplasma gondii.