目的探讨紫金标抗单纯疱疹Ⅰ型病毒(HSV-1)的分子机制,为进一步开发该药提供理论依据。方法采用不同剂量的紫金标作用于适量HSV-1感染的Vero细胞,以50%组织细胞感染量(TCID₅₀),细胞病变效应(CPE),蚀斑形成单位(PFU)为评价指标,采用CsCl密度梯度离心,RT-PCR,Western Blot以及扫描电镜和透射电镜为研究手段。结果紫金标无直接灭活HSV-1的作用,也不能影响病毒的释放;但可干扰HSV-1对宿主细胞的吸附,选择性地抑制HSV-1DNA及蛋白质合成,不同浓度的紫金标能明显抑制HSV-lgD基因mRNA表达。结论紫金标能抑制HSV-1对宿主细胞的吸附选择性地抑制HSV-1DNA及蛋白质合成,抑制HSV-1gD基因转录。

OBJECTIVE To study mechanisms of the extract from Ceratostigma willmattianum against herpes simplex virus type 1(HSV-1) in vitro . METHODS C. willmattianum in various concentrations was applied to Vero cells infected by HSV-1. Fifty percent tissue culture infective dose (TCID₅₀), cytopathic effect (CPE), plaque forming unit (PFU) were used as estimating index for antiviral activity. DNA synthesis was determined by isopycnic separation of cellular and viral DNA in CsCl gradients centrifugation. RT-PCR were used as estimating HSV-1 gD mRNA expression. Protein synthesis was determined by SDS-PAGE and Western Blot. RESULTS The extract from the plant neither killed HSV-1 directly nor released it from the infected cells, it showed interfering effect on the absorption of HSV-1 to Vero cells, which was counted for the observed inhibitory HSV-1 infectivity, and inhibiting HSV-1 gD mRNA expression. The above concentrations of C. willmattianum did not show any inhibitory effects on cellular protein synthesis, but selectively inhibited the viral protein synthesis. CONCLUSIONS C. willmattianum possesses strong anti-HSV-1 activity in vitro. The possible mechanisms include virus absorption,HSV-1 gD gene transcription, and selectively inhibitory effects on HSV-1 DNA and protein synthesis.