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目的:研究大蒜新素对人巨细胞病毒(HCMV)即刻早期(ie)、早期(e)和晚期(l)基因在转录水平的影响,探讨大蒜新素抗HCMV效应的作用机制。方法:建立HCMV AD169株(MOI=2.5)感染细胞和大蒜新素(9.6 mg.L-1)处理感染细胞模型,并用相应剂量(2.3 mg.L-1)的更昔洛韦(GCV)作比较,用实时荧光定量PCR方法检测各组细胞感染后0.5,2,4,6,12,24 h病毒ul122,ul123,ul54,ul83 mRNA水平的动态变化。结果:大蒜新素处理组ul122,ul123 mRNA的表达量始终明显低于感染对照组(P<0.05),而更昔洛韦处理组ul122 mRNA在0.5～6 h与病毒对照组无明显差异。大蒜新素对AD169ul122,ul123 mRNA的抑制率在感染后24 h分别为75.2%,70.4%。2药物处理组ul54 mRNA表达量始终低于病毒对照组(P<0.05),大蒜新素和更昔洛韦对ul54 mRNA的抑制率在感染后24 h分别为45.4%,27.2%。在感染后6 h各组ul83 mR-NA表达明显增多,以病毒感染对照组变化最为明显。大蒜新素和更昔洛韦对ul83 mRNA的抑制率在感染后24 h分别为45.9%,26.2%。结论:大蒜新素可显著抑制HCMVAD169毒株ie基因(ul122和ul123)的转录,导致其mRNA表达明显降低,对e基因(ul54)和l基因(ul83)转录水平亦有所抑制,表明病毒ie基因可能是大蒜新素抗HCMV作用的主要环节。
OBJECTIVE: To investigate the effect of allitridin on the transcriptional level of early, early and late genes of human cytomegalovirus (HCMV), and to explore the mechanism of allitridin against HCMV. Methods: The transfected cells were infected with HCMV AD169 (MOI = 2.5) and allitridin (9.6 mg.L-1), and treated with ganciclovir (GCV) at the corresponding dose (2.3 mg.L- The dynamic changes of the mRNA levels of ul122, ul123, ul54 and ul83 at 0.5, 2, 4, 6, 12 and 24 h after infection were detected by real-time fluorescence quantitative PCR. Results: The expression of ul122 and ul123 mRNA in allitridin treatment group was always lower than that in infected control group (P <0.05). However, ul122 mRNA in ganatoxin treatment group had no significant difference with virus control group at 0.5-6 h. Allitridin AD169ul122, ul123 mRNA inhibition rate of 24 h after infection were 75.2%, 70.4%. The expression of ul54 mRNA in the drug-treated group was always lower than that in the virus control group (P <0.05). The inhibitory rates of allitridin and ganciclovir on ul54 mRNA were 45.4% and 27.2% at 24 h after infection respectively. At 6 h after infection, the expression of ul83 mR-NA increased significantly in each group, with the most obvious change being in the control group infected with virus. The inhibitory rates of allitridin and ganciclovir on ul83 mRNA were 45.9% and 26.2% at 24 h after infection, respectively. CONCLUSION: Allitridin can significantly inhibit the transcription of ie gene (ul122 and ul123) of HCMVAD169 strain, resulting in a significant decrease of its mRNA expression, as well as inhibition of transcription of ul54 gene and ul83 gene, indicating that virus ie Gene may be allicin anti-HCMV the main link.