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目的研究真菌菌丝体提取物对人巨细胞病毒基因转录的抑制作用,探讨其抗HCMV效果及其机制。方法建立抗HCMV细胞模型,用半定量PCR检测细胞感染后不同时间UL122、UL123、UL44、UL57、UL86mRNA表达量的变化。结果 UL122、UL123mRNA的表达高峰为HCMV感染细胞后24h,UL44、UL57mRNA的表达高峰为感染后96h,UL86mRNA的表达高峰为感染后120h。真菌菌丝体提取物与HCMV作用后接种细胞及HCMV接种细胞后加入真菌菌丝体提取物24h检测UL122、UL123mRNA的表达量均显著降低。而病毒完成穿入过程后加入50μg/ml真菌菌丝体提取物与加入5μg/ml组相比UL122、UL123mRNA表达量更少。HCMV进入细胞6h后加入真菌菌丝体提取物再培养96h,UL44、UL57mRNA的表达量无显著变化。HCMV进入细胞24h后加入真菌菌丝体提取物再培养120h,UL86mRNA的表达表达量无显著变化。结论真菌菌丝体提取物可显著抑制HCMV AD169毒株IE基因(UL122、UL123)的转录,导致其mRNA表达显著降低,且抑制作用存在量效关系;但对E基因(UL44、UL57)和L基因(UL86)转录水平抑制作用不明显,表明真菌菌丝体提取物抗HCMV的重要作用机制是通过抑制HCMV IE基因的转录和翻译,进而抑制病毒的复制增殖。
Objective To study the inhibitory effect of fungal mycelium extract on human cytomegalovirus (CMV) gene transcription and to explore its anti-HCMV effect and its mechanism. Methods The anti-HCMV cell model was established. The changes of UL122, UL123, UL44, UL57 and UL86 mRNA expressions were detected by semi-quantitative PCR at different times after infection. Results The expression of UL122 and UL123 mRNA peaked at 24h after infection with HCMV. The peak expression of UL44 and UL57 mRNA was 96h after infection, and the peak expression of UL86 mRNA was 120h after infection. After fungal mycelium extract and HCMV were inoculated into cells and HCMV inoculated cells, the mycelial extracts of fungi were added for 24h to detect the expression of UL122, the expression of UL123 mRNA was significantly reduced. The addition of 50μg / ml fungal mycelium extract after the completion of the virus into the process of UL122, UL123 mRNA expression less than adding 5μg / ml group. HCMV into the cells 6h after adding fungal mycelium extract cultured 96h, UL44, UL57 mRNA expression no significant change. HCMV into the cells 24h after fungal mycelium extract was cultured 120h, UL86mRNA expression was no significant change. Conclusion The mycelial extracts of fungi can significantly inhibit the transcription of IE gene (UL122 and UL123) of HCMV AD169 strain, resulting in a significant decrease in mRNA expression and the dose-response relationship of E6 gene (UL44, UL57) and L The inhibitory effect of UL86 transcription was not obvious, indicating that the important mechanism of fungal mycelium extract anti-HCMV inhibition of viral replication and proliferation by inhibiting the transcription and translation of HCMV IE gene.