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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 clarify the mechanism of anti-rotavirus in vitro treated by Wang Lian Lian Pu Yin Plus Talc, Scutellaria baicalensis and Indadin-4 Decoction, and to observe the effect of anti-rotavirus diarrhea animal model on the animal model, providing a scientific basis for clinical application. METHODS: Toxic effects of a Mongolian medicine and Chinese herbs on MA-104 cells (rhesus monkey embryonic kidney cells) were tested. MTT (4,5-dimethylthiazole-2,5-diphenyltetrazolium bromide) method was established according to Takeuchi To detect the cytotoxicity of the drug. To prevent the virus from the virus infection, monolayers were pretreated with different concentrations of drugs for 2 h and then 100 TCID50 / 0.1 mL (Tissue culture infective dose) 0.1 mL, adsorption 2 h, the virus was discarded, PBS (Phosphate Buffer Saline, phosphate buffer) was washed twice with cell maintenance fluid culture, observed under inverted microscope cell changes every day; the direct effect of the drug on the virus, Different concentrations of drugs and 100TCID50 / 0.1 mL of the virus mixture mixed for 2 h, and then infected monolayer cells, adsorption 2 h, the supernatant was removed, cell maintenance solution was added to continue the culture, the cells were observed daily under an inverted microscope The cells were infected with 100TCID50 / 0.1 mL of virus at 0.1 mL per cell for 2 h and then the virus was taken in. Different concentrations of drug-containing maintenance solution were added to the cells. Watch the cells Of. In Experiment 2, a mouse model of diarrhea caused by senna was replicated. The effects of Mongolian medicine, traditional Chinese medicine and atropine on diarrhea were compared. The wet diarrhea was observed and the degree of inhibition of the drug on diarrhea was observed. Results: The TC50 (half cytotoxic concentration) of Mongolian medicine was 2.76 mg / mL and the TC50 of Chinese medicine was 24.35 mg / mL. The toxicity of Mongolian medicine to MA-104 was obviously higher than that of traditional Chinese medicine. The main action of traditional Chinese medicine against rotavirus was to inhibit the growth of virus However, it is less effective in blocking virus adsorption and direct killing. Mongolian medicine can significantly inhibit the replication of rotavirus, which has a good effect in blocking virus adsorption, direct killing and inhibiting virus proliferation. However, cytotoxicity Large; Chinese medicine low-dose group and Mongolian medicine high-dose group anti-diarrhea better than other groups.