外感清热解毒方抗呼吸道病毒活性的体外实验研究

Effect of Waigan Qingre Jiedu Formula Against Respiratory Virus In Vitro

目的:研究外感清热解毒方体外抗呼吸道病毒的活性。方法:体外培养MDCK、HEL和A549细胞,分别接种流感病毒H1N1、腺病毒AV1和呼吸道合胞病毒RSV1,给予外感清热方进行干预,以病毒唑作为对照。采用MTT法检测药物对细胞的毒性和对病毒的治疗指数,采用红细胞凝集实验和观察细胞病变(CPE)进行分析,研究该方抗呼吸道病毒活性。结果:外感清热方对H1N1的IC 50为0.028 mg/m L,TI为2.07。预防给药在0.01 mg/m L时红细胞凝集实验为阴性,表明该方能够阻断H1N1病毒入侵,有预防作用。治疗给药在0.01 mg/m L时红细胞凝集实验为阴性,表明该方对病毒有治疗作用。直接杀伤实验该方能够降低病毒滴度1个梯度。在0.005 mg/m:时抗AV1红细胞凝集实验为阴性,表明该方能够抑制AV1。镜下观察CPE表明0.01 mg/m L以上浓度的中药能够抑制RSV1。结论:外感清热解毒方具有较好的抗H1N1、AV1和RSV1的活性,具有高效低毒的特点。

Objective： To study the activity of Waigan Qingre Jiedu decoction against respiratory virus in vitro. Methods： MDCK,HEL and A549 cells were cultured in vitro; influenza virus H1N1,adenovirus AV1 and respiratory synthetic virus RSV1 were inoculated respectively. Waigan Qingre Jiedu decoction was given to intervene,and Ribavirin was given as control. MTT method was used to detect the cytotoxicity of the drug and therapeutic index. Hemagglutination test and Cytopathic effect were used to study the effect against respiratory virus. Results： IC50 of Waigan Qingrejiedu decoction against H1N1 was 0. 028 mg/m L. TI was 2. 07. In the prevention administration experiment,hemagglutination test at the concentration of 0. 01 mg/m L was negative,which indicates that the decoction could block H1N1 virus invasion and has preventive effect. In the therapy administration experiment,hemagglutination test at the concentration of 0. 01 mg/m L was negative,which showed that the decoction had a therapeutic effect on the virus. In the Direct killing experiment,the decoction can reduce the virus titer by 1 gradient. Hemagglutination test against AV1 at the concentration of 0. 005 mg/m L was negative,which showed that the decoction could inhibit AV1. The observation of Cytopathic effect under the microscope showed that the decoction could inhibit RSV1 in the concentration of 0. 01 mg/m L. Conclusion：Waigan Qingre Jiedu decoction has high activity against H1N1,AV1 and RSV1,with the advantage of having high efficiency and low toxicity.