Isolation and characterization of an anti-HSV polysaccharide from Prunella vulgaris
Isolation and characterization of an anti-HSV polysaccharide from Prunella vulgaris
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DOI：
10.1016/s0166-3542(99)00053-4
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发表时间：
1999-11-01
影响因子：
7.6
通讯作者：
Blay, J
中科院分区：
文献类型：
--
作者：
Xu, HX;Lee, SHS;Blay, J
采用热水提取、乙醇沉淀、凝胶渗透柱层析等方法，从夏枯草中分离得到一种水溶性物质。化学测试表明，该物质为阴离子多糖。空斑减少试验表明，100mU g/ml的多糖对单纯疱疹病毒1型和2型(HSV-1和HSV-2)有活性，但对巨细胞病毒、人类流感病毒A和B型、脊髓灰质炎病毒1型和水泡性口炎病毒没有活性。该多糖对HSV-1和HSV-2的半数空斑减少量为10微克/毫升，对HSV-1和HSV-2临床分离株和已知的阿昔洛韦耐药株(TK缺陷株或聚合酶缺陷株)的抑制作用相似。将HSV-1与该多糖在4℃、25℃或37℃预先孵育，可完全消除HSV-1的感染性，但用该多糖预处理Vero细胞并不能保护细胞免受病毒感染。在HSV-1感染Vero细胞后0、2、5.5和8h加入MOI为5的多糖组，20h细胞内感染性病毒产量分别下降100、99、99和94%。相比之下，类似的肝素添加分别显示细胞内病毒产量下降85%、63%、53%和3%。这些结果表明，该多糖可能通过竞争细胞受体以及病毒穿透细胞后的一些未知机制来抑制HSV。夏枯草多糖对哺乳动物细胞无细胞毒作用，最高浓度为0.5 mg/ml，未显示任何抗凝血活性。综上所述，从普通拟青霉中分离得到的多糖具有抗单纯疱疹病毒的特异性，其作用方式与肝素等其他阴离子碳水化合物不同。(C)1999 Elsevier Science B.V.保留所有权利。
A water soluble substance was isolated from a Chinese herb, Prunella vulgaris, by hot water extraction, ethanol precipitation and gel permeation column chromatography. Chemical tests showed that the substance was an anionic polysaccharide. Using a plaque reduction assay, the polysaccharide at 100 mu g/ml was active against the herpes simplex virus types 1 and 2 (HSV-1 and HSV-2), but was inactive against cytomegalovirus, the human influenza virus types A and B, the poliovirus type 1 or the vesicular stomatitis virus. The 50% plaque reduction dose of the polysaccharide for HSV-1 and HSV-2 was 10 mu g/ml. Clinical isolates and known acyclovir-resistant (TK-deficient or polymerase-defective) strains of HSV-I and HSV-2 were similarly inhibited by the polysaccharide. Pre-incubation of HSV-1 with the polysaccharide at 4, 25 or 37 degrees C completely abrogated the infectivity of HSV-1, but pre-treatment of Vero cells with the polysaccharide did not protect cells from infection by the virus. The addition of the polysaccharide at 0, 2, 5.5 and 8 h post-infection of Vero cells with HSV-1 at a multiplicity of infection (MOI) of five reduced the 20 h-yield of intracellular infectious virus by 100, 99, 99 and 94%, respectively. In contrast, a similar addition of heparin showed 85, 63, 53 and 3% reduction of intracellular virus yield, respectively. These results suggest that the polysaccharide may inhibit HSV by competing for cell receptors as well as by some unknown mechanisms after the virus has penetrated the cells. The Prunella polysaccharide was not cytotoxic to mammalian cells up to the highest concentration tested, 0.5 mg/ml and did not show any anti-coagulant activity. In conclusion, the polysaccharide isolated from P. vulgaris has specific activity against HSV and its mode of action appears to-be different from other anionic carbohydrates, such as heparin. (C) 1999 Elsevier Science B.V. All rights reserved.