三种方法检测和评价美洲商陆叶和果的急性毒性

Using three methods to test and evaluate the fruit and leaf's toxicity of Phytolacca Americana L.

目的分别使用经典的小鼠急性经口毒性试验、体外细胞毒性试验及线虫毒性试验三种方法对美洲商陆叶子和果实进行急性毒性检测和评估。方法小鼠急性经口毒性试验中,美洲商陆叶使用一次最大限量法,美洲商陆果使用霍恩氏法;体外细胞毒性试验使用CHL(中国仓鼠肺细胞)中性红染色法;线虫毒性试验采用96孔板对同步化的秀丽隐杆线虫L4期幼虫进行24 h染毒。结果小鼠急性经口毒性试验表明美洲商陆叶的小鼠经口最大耐受剂量(MTD)≥20.00 g/kg BW,为无毒级,而果的LD50>10.00 g/kg BW,为实际无毒,果的小鼠急性毒性大于叶的毒性;在细胞毒性试验中,叶和果的IC50分别为7.4和5.6μg/ml;在线虫毒性试验中,经215.0 mg/ml的美洲商陆叶染毒24 h后,仍未出现死亡,而果的LC50=16.5 mg/ml。结论经典小鼠和线虫染毒模型均显示果的毒性大于叶的毒性;在细胞毒性试验中,虽然果的细胞毒性略大于叶的毒性,但差别比较微弱。提示线虫模型比细胞模型更具有毒性预筛的潜在应用价值。

Objective The fruit and leaf＇s toxicity of Phytolacca Americana L. were tested and evaluated by acute oral toxicity test in mouse, in vitro cytotoxicity test and C. elegans toxicity test. Methods In the experiment of acute oral toxicity test in mouse, the maximum limit method was used to test the leafs toxicity and Horn＇s method to test the fruit＇s toxicity. In vitro cytotoxieity test was carried out on CHL cells using neutral red uptake method. C. elegans toxicity test was carried out after 24 h exposure using 96-well plates. Results By acute oral toxicity test in mouse, it was proved that the leaf was notoxic with the maximum tolerated dose no less than 20. 00 g/kg BW and the fruit was actually no-toxic with LD50 greater than 10. 00 g/kg BW. The fruit＇s toxicity was greater than the leaf＇s. In the in vitro cytotoxicity test, the leaf and the fruit＇s IC50 were 7.4 and 5.6 μg/ml respectively. In C. elegans acute toxicity test, the death of the nematodes was not detected at the dose of 215.0 mg/ml after 24 h exposure to leaf, whereas the IC50 of fruit was detected as 16.5 μg/ml. Conclusion It was indicated that the Phytolacca Americana L. fruit＇s toxicity wasgreater than the leafs using classic mouse and C. elegans exposure model. In the in vitro cytotoxicity model, the fruit＇s toxicity was a little higher than the leaf＇s with minor difference. It was suggested that C. elegans model had more practical potential than in vitro cytotoxicity model on toxicity screening.