复方中药制剂体外杀纤毛虫及毒性试验研究

In Vitro Killing of Ciliates and Toxicity of Compound Chinese Herbal Medicines to Turbot Scophthalmus maximus

研究3个复方中草药水煎浓缩液(1 g/mL)(复方Ⅰ:十大功劳、黄柏、半枝莲、苦参、百部、槟榔;复方Ⅱ:大黄、黄芩、野菊花、蛇床子、石榴皮、乌梅;复方Ⅲ:地丁、蒲公英、野菊花、地锦草、败酱草、龙胆草、鱼腥草、青蒿)体外杀水滴伪康纤虫和海洋尾丝虫的效果及对体质量18~20 g大菱鲆的毒性,旨在筛选出高效、低毒、少残留的治疗盾纤毛虫病的中药组合。体外杀虫试验在96孔细胞培养板的每孔中加入90μL药液和10μL离心后的虫液,药物剂量分别为0、5、10、15、20、25、30 mg/kg,每组3个平行,显微镜下观察盾纤毛虫的状态。体外杀虫试验结果显示,复方Ⅰ的剂量在0~30 mg/kg时对海洋尾丝虫无杀害作用;复方Ⅲ仅在药物剂量为30 mg/kg时对海洋尾丝虫有杀害作用,其他剂量时无效;复方Ⅱ20、25、30 mg/kg组1 h水滴伪康纤虫全部死亡,30 mg/kg组3 h海洋尾丝虫全部死亡。随即在循环水箱中进行为期7 d的大菱鲆毒性试验,随机分为4组,即空白对照组(0 mg/kg)、推荐剂量组(20 mg/kg)、3倍剂量组(60 mg/kg)、5倍剂量组(100 mg/kg),每组3个平行。试验结束时,鱼尾静脉采集全血和血清,使用血液细胞分析仪和全自动生化分析仪检测血常规和血液生化指标。大菱鲆毒性试验结果显示,复方Ⅱ药浴后,大菱鲆血液中白细胞、嗜酸性细胞等血常规指标以及葡萄糖、球蛋白含量等血液生化指标均无显著差异;各试验组大菱鲆的鳃、肠道、胃、肝脏和脾脏均无明显的病理变化。

Ciliates Pseudocohnilemus persalinus and Uronema marinum were held in a 96-well cell culture plate containing 90μL of solution of 3 compound Chinese herbal medicines including CompoundⅠ:Mahonia,Phellodendron,Scutellaria barbata,Sophora flavescens,Stemona,and Areca;CompoundⅡ:Rheum,Scutellaria,Chrysanthemum indicum,Fructus cnidii,pomegranate peel,and mume;and CompoundⅢ:dice,dandelion,chrysanthemum,broccoli,Patrinia,Gentian,Houttuynia,and Artemisia annua,at a dose of 0,5,10,15,20,25,and 30 mg/kg,and 10μL of centrifuged ciliates solution containing the ciliates at a density of 10~3 individuals/mL to screen the Chinese herbal medicine formulas in vitro insecticidal test by observation of the ciliates under a light microscope.Blood biochemical and histopathological indices were determined and observed in juvenile turbot Scophthalmus maximus exposed to the 3 compound Chinese herbal medicines above at a dose of 0,20,60,and 100 mg/kg for 7 days to evaluate toxicity of the 3 compound Chinese herbal medicines to turbot.Killing effect was observed in U.marinum exposed to CompoundⅢat a dose of 30 mg/kg,without mortality in Compound I solution at a dose of 0—30 mg/kg.All ciliate P.persalinus was found to be dead in the compoundⅡgroup for 1 hour at a dose of 20,25,and 30 mg/kg,and U.marinum was all dead in the compoundⅡgroup for 3 hour at a dose of 30 mg/kg.There were no significant differences in blood routine indicators including white blood cells and eosinophils,as well as blood biochemical indicators such as glucose and globulin,without obvious pathological and pathological changes in the intestine,stomach,liver and spleen in each test group.