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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)10~80μg/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)40μg/ml、80μg/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;160μg/ml的大蒜素对细胞有明显的毒副作用。(3)大蒜素80μg/ml时,台盼兰着色率最高,弓形虫的活力最低。(4)大蒜素80μg/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80μg/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。
Objective To observe the effect and mechanism of garlic extract against Toxoplasma gondii in vitro. Methods The tachyzoites of RH strain Toxoplasma gondii RH strain were co-cultured with rabbit kidney cells. Different concentrations of allicin (experimental group) and sulfadiazine (positive control group) were added. After being cultured for different time, cells were removed. Toxoplasma gondii tachyzoites per tuber of the number of tachyzoites; the use of MTT colorimetric Toxoplasma gondii tachyzoites invasion of cells and normal cells proliferation; observed by trypan blue allicin on Toxoplasma gondii Tachyzoites. The apoptosis rate of Toxoplasma gondii Tachyzoites was detected by TUNEL end-labeling method, and the effect and mechanism of the Toxoplasma gondii Toxoplasma gondii tachyzoites were discussed. Results (1) Allicin at 10 ~ 80μg / ml could resist Toxoplasma gondii infection in a dose-dependent manner, with no significant correlation with time. (2) 40μg / ml, 80μg / ml allicin can not inhibit cell proliferation, no significant toxic effects on the cells; 160μg / ml allicin has obvious toxic and side effects on the cells. (3) Allicin 80μg / ml, trypan blue maximum coloring rate, Toxoplasma gondii minimum activity. (4) The concentration of allicin 80μg / ml, Toxoplasma tachyzoites apoptosis rate was the highest. Conclusion Allicin can inhibit tachyzoites Toxoplasma gondii activity, the host cell infectivity, proliferation in the cell, no significant side effects, the most appropriate concentration of 80μg / ml. Allicin is a good anti-Toxoplasma gondii anti-Toxoplasma gondii apoptosis mechanism induced apoptosis.