大蒜新素抑制人巨细胞病毒即刻早期抗原表达的实验研究

An Experimental Study on the Effect of Allitridin on Inhibiting the Expression of HCMV Immediate-early Antigens in Vitro

目的 :通过研究大蒜新素对人巨细胞病毒即刻早期抗原 (HCMVIEA)的抑制作用 ,探讨大蒜新素体外对HCMV感染的预防、阻断和治疗效应及其机制。方法 :采用细胞形态观察和MTT比色法检测大蒜新素的细胞毒性。以流式细胞术定量检测和比较大蒜新素不同剂量 ,不同处理时间 (2 4 ,4 8,72 ,96h)以及病毒感染不同阶段 (病毒吸附前、吸附时和吸附后 )应用大蒜新素时HCMVIEA表达水平变化。结果 :人胚肺成纤维细胞对大蒜新素的最大耐受浓度为 9.6 0mg·L-1。病毒吸附后持续用药可显著抑制HCMVIEA的表达 ,用药 72hHCMVIEA抑制率达 89.3% ;大蒜新素预处理细胞后感染病毒 ,HCMVIEA抑制率为 2 8.6 % ;药物与病毒吸附同时应用 ,即刻早期抗原 (IEA)抑制率仅为 10 .3%。结论 :大蒜新素体外持续用药可显著抑制HCMVIEA的表达 ;其预防作用较弱 ;即时抑制效应不明显。大蒜新素对IEA表达的明显抑制可能是大蒜新素抗HCMV效应的主要作用机制之一。

Objective: To investigate the prophylactic,blocking and therapeutic effects of Allitridin on inhibiting HCMV proliferation by measuring the expression level of HCMV IEA in vitro and explore the mechanism of Allitridin anti-HCMVactivity. Methods: The cytotocity of Allitridin was evaluated through MTT colorimetry and cell morphology.HCMV IEA levels were quantitatively detected by Flow Cytometry respectively under the following conditions:Allitrdin was given before(pretratcd for 24 h),during,or after viral inoculation in which a serial doses(maximum tolerant concentration,MTC for human embyo lung cells,HEL) of Allitridin was used to treat HCMV infected HLE cells for different durations (24,48,72,96 h) after viral infection. Result: The MTC of Allitridin was 9.60 mg·L -1 .Allitridin remarkably inhibited the expression of HCMV IEA in vitro .Within MTC,the inhibitory rate had a significative correlation with its dosage ( r =0.96).At the time of IEA highest expression (72 h after iinfection),inhibitory effect was the greatest(inhibitory rate:89.3%).With pretreatment of Allitridin,the inhibitory rate was 28.6%. When Allitridin was used together with HCMV inoculation,IEA inhibitory rate was only 10.3%. Conclusion: Allitridin can inhibit HCMV, IEA expression in vitro remarkably which is probably one of the major mechanisms of Allitridin anti-HCMV activity because IEAs are the very important regulatory factors for the expression of all HCMV genes.Its therapeutic effect is the best at the peak stage of IE1 gene expression (72 h after infection)but it has low prophylactic and little blocking effect.