地龙抗呼吸道合胞病毒的有效部位筛选  

Screening for Anti-Respiratory Syncytial Virus Active Parts of Geosaurus

作　　者：张加泽 任莹[1] 王清[1] 候宝山 牛文斐 姜爱雯[2] 周长征[1] 

机构地区：[1]山东中医药大学,山东济南250355 [2]河北北方学院附属第一医院,河北张家口075000

出　　处：《中国现代中药》2016年第9期1125-1128,共4页Modern Chinese Medicine

基　　金：山东省中医药抗病毒应用基础与关键技术及产业化课题(XTCX2014C01-04)

摘　　要：目的:研究地龙不同极性的粗提取物经纯化分离后对呼吸道合胞病毒(RSV)的体外抗病毒活性,从而确定抗病毒的有效部位。方法:预实验依次用乙酸乙酯、95%乙醇水溶液、水作为溶剂提取药物,采用RSV-Hep-2细胞感染模型,初步检测地龙的体外抗RSV活性。依据预实验的结果,分别采用水提醇沉法和醇提水沉法对粗提取物进一步地分离和纯化,将处理后的上清液和沉淀物分别进行体外抗病毒实验。然后根据实验结果将醇提水沉后的沉淀过大孔树脂DM130,分别用25%、50%、75%的乙醇水溶液洗脱,将洗脱液浓缩后做抗病毒实验,在显微镜下观察细胞的病变,用酶标仪测定中性红染色A540值,计算其治疗指数。结果:乙酸乙酯、乙醇、水3种粗提取物中水粗提取物对呼吸道合胞病毒的抑制效果最好,TI=74.21。将粗提取液水提醇沉或醇提水沉后发现醇提水沉的沉淀对呼吸道合胞病毒的抑制效果最好,TI=107.11。将地龙醇提水沉的沉淀过大孔树脂DM130发现75%乙醇2号水溶液冲洗的洗脱液抑制呼吸道合胞病毒的效果最好,TI=63.05.结论:地龙的醇溶性成分对RSV病毒具有明显的抑制作用。Objective： To study anti-RSV activities of the crude extracts with different polarity after purified and separated from the Geosaurus, determine the effective parts of antivirus further. Methods： Ethyl acetate, 95% ethanol and pure water were used as the solvent to extract drugs respectively. Anti-RSV activities in vitro were detected preliminarily by RSV-Hep-2 virus-cell infection model of ethanol. Crude extracts were purified and separated further by water extraction ethanol precipitation and ethanol extraction water precipitation methods respectively. Supernatants and sediments were used as the antivirus experiments in vitro. After the sediments from ethanol extraction water precipitation DM were filtered by 130 macroporous absorbent resin, the filters were eluted by 25%, 50%, 75% ethanol respectively according to the results. In the end the antivirus experiments were carried out by the concentrated eluent. The cytopathic effect （CPE） was observed under a microscope. Neutral red staining was measured in a microplate reader at 540 nm to calculate the therapeutic index （TI）. Results： Among the three total extracts from Geosaurus by ethyl acetate, 95% ethanol and pure water, the pure water total extract was the most effective for Anti RSV（TI = 125. 72）. The filters from sediments from ethanol extraction water precipitation DM were filtered by 130 macroporous absorbent resin, and it showed that eluent of 75% ethanol in number 2 was the most effective for Anti RSV（ TI = 63.05 ）. Conclusion： The ingredients of alcohol dissolved have the most significant inhibited effect on respiratory syncytial virus.

关 键 词：地龙 粗提取物 水提醇沉 醇提水沉 大孔树脂 抗呼吸道合胞病毒 

分 类 号：R285.5[医药卫生—中药学]