18味中药提取物体外抗鸭瘟病毒活性的研究

Study on antivirus activity of crude extract from eighteen kinds of traditional Chinese medicine against duck plague virus in vitro

我国传统中药在临床中广泛用于抗病毒疾病的治疗,该研究以18味传统中药为研究对象,希望找到对鸭瘟病毒具有良好抑制作用的中药原料。首先,通过细胞病变观察法(cytopathic effect,CPE)初步筛选了18味中药水提取物和醇提物进行体外抗鸭瘟病毒活性研究,发现当虎杖醇提物质量浓度为0.062 5mg/mL和0.031 3mg/mL,茜草醇提物质量浓度为0.007 8mg/mL时,可抑制鸭瘟病毒引起的鸭胚成纤维细胞病变。进而通过四甲基偶氮唑盐法[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide,MTT]进一步研究了虎杖和茜草醇提物的体外抗鸭瘟病毒活性。结果显示:当虎杖质量浓度为0.062 5mg/mL、茜草质量浓度为0.007 8mg/mL时,对鸭瘟病毒的体外抑制率分别为76.50%、48.91%,虎杖醇提物的半数有效质量浓度(50%inhibitory concentration,IC50)为0.022 7mg/mL,茜草醇提物的IC50>0.007 8mg/mL。提示虎杖和茜草醇提物均有不同程度的体外抗鸭瘟病毒活性,其中虎杖效果优于茜草,具有进一步研究的价值。

Summary Traditional Chinese medicine is rich in resources and has the characteristics of low prices and low-toxic side effects, which is widely used in the treatment of animal diseases, including viral diseases. Duck plague virus （DPV） is an acute, contagious herpesvirus infection of duck, geese, and swans of all ages and species. This disease has been responsible for significant economic losses in domestic and wild waterfowl as results of mortality, and decreased egg production. There is no effective drug on the therapy of this virus disease. We attempted to screen out a few of effective traditional Chinese medicines on DPV from eighteen kinds of traditional Chinese medicines. The methods were as follows. At first, the cytotoxic effect of the water extracts and alcohol extracts of the traditional Chinese medicines on DEFs monolayer were determined by the method of cytopathic effect （CPE）. All the concentrations of extracts used in antiviral activity assays were below the maximal non-cytotoxic concentration （MNCC）. In the antiviral activity assay, the method of CPE was used in preliminary screening for the effective extracts. The virus inhibitory ratios of the effective extracts against DPV were tested by the method of [3-（4,5-dimethylthiazol 2 yl）-2,5- diphenyltetrazolium bromide, MTT] colorimetric analysis. The results showed that the virus inhibitory ratios against DPV were 76.50% and 48.91% with the concentrations 0. 062 5 mg/mL and 0. 007 8 mg/mL of Rhizorna polygoni cuspidati and Radix rubiae. Additionally, the 50% inhibitory concentration of Rhizoma polygoni cuspidati and Radix rubiae were 0. 022 7 mg/mL and greater than 0. 007 8 mg/mL respectively. The result above indicated that the alcohol extracts of Rhizoma polygoni cuspidati and Radix rubiae showed the antiviral activities at varying degrees in vitro and the Rhizoma polygoni cuspidati, whose antiviral activity is better than Radix rubiae, might be a potential agent in the prevention of DPV infections.