Aguilar J S, Roy D, Ghazal P, Wagner E K
Dept. of Mol. Biol. & Biochem, U. Calif. Irvine, 19172 Jamboree Road, Irvine, CA 92697, USA.
BMC Infect Dis. 2002 May 24;2:9. doi: 10.1186/1471-2334-2-9.
Dimethyl sulfoxide (DMSO) is frequently used at a concentration of up to 95% in the formulation of antiherpetic agents because of its properties as a skin penetration enhancer. Here, we have analyzed the effect of DMSO on several parameters of Herpes Simplex Virus replication.
Productive infection levels of HSV-1 were determined by plaque assay or by reporter gene activity, and its DNA replication was estimated by PCR. Transcript levels were evaluated with HSV-specific DNA micro-arrays.
DMSO blocks productive infection in vitro in different cell types with a 50% inhibitory concentration (IC50) from 0.7 to 2% depending upon the multiplicity of infection. The concentration dependence exhibits a Hill coefficient greater than 1, indicating that DMSO blocks productive infection by acting at multiple different points (mechanisms of action) with positive cooperativity. Consistently, we identified at least three distinct temporal target mechanisms for inhibition of virus growth by DMSO. At late stages of infection, DMSO reduces virion infectivity, and markedly inhibits viral DNA replication. A third mode of action was revealed using an oligonucleotide-based DNA microarray system for HSV. These experiments showed that DMSO reduced the transcript levels of many HSV-1 genes; including several genes coding for proteins involved in forming and assembling the virion. Also, DMSO markedly inhibited some but not all early transcripts indicating a previously unknown mode for inhibiting the early phase of HSV transcription-replication cycle.
These observations suggest that DMSO itself may have a role in the anti-herpetic activity of formulations utilizing it as a dispersant.
二甲基亚砜(DMSO)因其作为皮肤渗透增强剂的特性,在抗疱疹药物制剂中常以高达95%的浓度使用。在此,我们分析了DMSO对单纯疱疹病毒复制的几个参数的影响。
通过蚀斑试验或报告基因活性测定HSV-1的生产性感染水平,并通过PCR估计其DNA复制。用HSV特异性DNA微阵列评估转录水平。
DMSO在体外阻断不同细胞类型中的生产性感染,50%抑制浓度(IC50)为0.7%至2%,具体取决于感染复数。浓度依赖性表现出大于1的希尔系数,表明DMSO通过在多个不同点(作用机制)以正协同作用发挥作用来阻断生产性感染。一致地,我们确定了DMSO抑制病毒生长的至少三种不同的时间靶点机制。在感染后期,DMSO降低病毒粒子的感染性,并显著抑制病毒DNA复制。使用基于寡核苷酸的HSV DNA微阵列系统揭示了第三种作用模式。这些实验表明,DMSO降低了许多HSV-1基因的转录水平;包括几个编码参与病毒粒子形成和组装的蛋白质的基因。此外,DMSO显著抑制了一些但不是所有的早期转录本,表明存在一种以前未知的抑制HSV转录-复制周期早期阶段的模式。
这些观察结果表明,DMSO本身可能在将其用作分散剂的制剂的抗疱疹活性中发挥作用。