红树林淡紫拟青霉胞外多糖体外抗HSV-1 的实验研究

In vitro anti-HSV-1 activity of extracellular polysaccharides from Paecilomyces lilacinuson isolated from Mangrove

摘要:

目的 探讨红树林淡紫拟青霉胞外多糖体外抗单纯疱疹病毒Ⅰ型（HSV-1）感染的活性，为开 发新型抗病毒药物提供实验基础。方法 以Vero 细胞为病毒感染靶细胞，以不同浓度淡紫拟青霉胞外多糖 作用于HSV-1 感染的各个阶段，以细胞病变程度测定病毒半数感染量，MTT 法检测淡紫拟青霉胞外多糖对 Vero 细胞的毒性作用、对HSV-1 的直接灭活作用、对病毒吸附和生物合成的影响。结果 淡紫拟青霉胞 外多糖对Vero 细胞的半数有毒浓度（CC50）为1 724.2μg/ml，该多糖浓度<900μg/ml 时，细胞存活率仍> 90%，且对细胞无增殖作用；在25 ～ 1 000μg/ml 浓度范围内，该多糖可在一定程度上抑制病毒吸附，并表 现出一定的量效关系，半数抑制浓度（IC50）为650.7μg/ml ；该多糖还具有抑制HSV-1 生物合成的作用， 其IC50 为547.7μg/ml，在25 ～ 1 000μg/ml 浓度范围内，抑制率与药物浓度呈量效关系，未发现该多糖对 HSV-1 有直接灭活作用。结论 淡紫拟青霉胞外多糖对Vero 细胞毒性较小，是一种非常安全的多糖；该多 糖具有一定的抗病毒作用，在一定浓度范围内可抑制HSV-1 吸附和生物合成，且表现出一定的量效关系。

Abstract:

Objective To investigate the anti-HSV-1 activity of the extracellular polysaccharides from Paecilomyces lilacinus isolated from Mangrove , so as to provide research basis for the development of new antiviral drugs. Methods The maxmium safe concentration of the extract to Vero cells was assessed by cytopathic effect (CPE) and MTT assay. The polysaccharides in different concentrations were applied to various phases of HSV-1 replication cycles respectively. The virus infectivity (TCID50) was examined by CPE, and the direct inactivation effect of the extract was observed by MTT assay, the inhibition effect of the extract on HSV-1 absorption and biosynthesis was examined by MTT assay too. Results The extracellular polysaccharides had little cytotoxic effect on Vero cells (CC50 = 1724.2 μg/ml). They could inhibit HSV-1 absorption on Vero cells at the dose of 25-1000 μg/ml to some extent in a dose-dependent manner (IC50 = 650.7 μg/ml). The biosynthesis of HSV-1 could be significantly inhibited by the extract at the dose of 25-1000 μg/ml in a dose-dependent manner (IC50= 547.7 μg/ml), but the extract couldn’t inactivate HSV-1 directly. Conclusions The extracellular polysaccharides are little harmful to Vero cells. They have conspicuous anti-HSV-1 activity, probably acting at the stage of HSV-1 binding to Vero cells and the virus biosynthesis stage in a dose-effect relationship.