中蒙医温病学王氏连朴饮加滑石、黄芩与茵达日-4味汤体外抗轮状病毒、抗腹泻实验  

Anti-Rotavirus and Anti-Diarrhea Trials of Mongolian Effective Prescription WANG's Lianpu Drink plus Talc and Skullcap; Yindari-4 Decoction in vitro

作　　者：闫腾飞[1] 韩雪梅[1] 金星[1] 

机构地区：[1]内蒙古医科大学,内蒙古呼和浩特010110

出　　处：《中华中医药学刊》2013年第9期1988-1991,I0008,共5页Chinese Archives of Traditional Chinese Medicine

基　　金：内蒙古自然科学基金资助项目(2010zd28)

摘　　要：目的:明确王氏连朴饮加滑石、黄芩与茵达日-4味汤体外抗轮状病毒的机制及观察其抗番泻叶所致腹泻动物模型的效果,为临床应用提供科学依据。方法:实验一蒙药和中药对MA-104细胞(恒河猴胚胎肾细胞)的毒性作用,根据Takeuchi建立MTT(4,5二甲基噻唑-2,5-二苯基四唑溴化物)法检测药物的细胞毒性作用;药物对病毒感染的预防作用,用不同浓度的药物预先处理单层细胞2 h后将100TCID50/0.1 mL(Tissue culture infective dose,半数组织培养感染剂量)的病毒每孔加入0.1 mL,吸附2 h,弃病毒液,PBS(Phosphate Buffer Saline,磷酸盐缓冲液)洗涤2次,加细胞维持液培养,每天于倒置显微镜下观察细胞变化;药物对病毒的直接杀伤作用,将不同浓度的药物与100TCID50/0.1 mL的病毒液等量混合作用2 h,再将其感染单层细胞,吸附2 h后,弃上清,加入细胞维持液继续培养,每天于倒置显微镜下观察细胞变化;药物对病毒穿入细胞后的抑制作用,先用100TCID50/0.1 mL的病毒每孔0.1 mL感染细胞,吸附2 h后,吸去病毒,加入不同浓度的含药维持液,每天于倒置显微镜下观察细胞变化。实验二复制番泻叶所致泄泻的小鼠模型,比较蒙药、中药与阿托品对腹泻的影响,通过计数湿粪,观察药物对腹泻的抑制程度。结果:蒙药的TC50(半数细胞毒性浓度)为2.76 mg/mL,中药的TC50为24.35 mg/mL,蒙药对MA-104细胞毒性明显大于中药;中药抗轮状病毒的主要作用方式为抑制病毒生物合成,但在阻断病毒吸附以及直接杀伤方面效果较差;蒙药能明显抑制轮状病毒复制,表现在阻断病毒吸附、直接杀伤以及抑制病毒增殖方面都有较好的效果,但细胞毒性较大;中药低剂量组和蒙药高剂量组抗腹泻效果优于其他组。Objective ： To understand the anti - rotavirus mechanism of WANGg Lianpu drink plus talc and skullcap as well as the Yindari - 4 Decoction and to observe the the effect of anti - diarrhea caused by senna in animal models, provi- ding a scientific basis for clinical application. Method： Experiment one：the experiment was toxic effects of Mongolian medicine and traditional Chinese medicine on the MA - 104 cells, according to Takeucbi, drug cytotoxicity was detected by MTI＇. Drugs on the prevention of viral infection ： pre - treatment with different concentrations of drugs was carried on for monolayer cells ,2 h later, 100TCIDs0/0.1 mL virus was added into holes with 0.1 mL for each, absorbing for 2 h, dis- posable virus solution was washed twice in PBS, adding cells to maintain the liquid culture, the cellular changes were ob- served under inverted microscope every day. Direct killing effect of drugs on the virus：different concentrations of drugs were mixed with 100TCID50/0. lmL virus solution with the equal quantity for 2 h. And then the supernatant was removed and added cells maintenance medium to culture and the cellular changes were observed under inverted microscope every. day. Drugs effect on inhibition of virus penetration：first 100TCIDS~/0.1 mL virus was used to infected cells with 0. l mL for each hole. 2 h adsorption later, the virus was removed, and drug - containing maintenance medium with difterent con- centrations was added, cells were observed daily under an inverted microscopic view. Experiment two：a mouse model of diarrhea induced by senna was established to compare the effect of Mongolian medicine and traditional Chinese medicine and atropine for diarrhea, by counting the wet manure to observe the degree of inhibition of drugs for diarrhea. Results： TCs0of Mongolian Medicine was 2.76 mg/mL and TCs0 of traditional Chinese medicine （TCM） was 24.35 mg/mL. Mon- golian Medicine has gTeater cell toxicity on the MA - 104 cells than traditional Chinese medicine. The main way of Chi- n

关 键 词：王氏连朴饮加滑石 黄芩 茵达日-4味汤 轮状病毒 体外 腹泻 

分 类 号：R291.2[医药卫生—民族医学]