真菌菌丝体提取物抑制人巨细胞病毒基因转录水平的研究  

Inhibition of human cytomegalovirus transcription by an extract of fungal mycelium

作　　者：张文利[1] 边贻海 马瑜[2] 李万美[1] 张华宁[3] 

机构地区：[1]山东省济宁卫生学校,山东济宁272100 [2]济南铁路疾病预防控制中心,山东济南250000 [3]山东省疾病预防控制中心,山东济南250062

出　　处：《中国病原生物学杂志》2016年第4期342-345,共4页Journal of Pathogen Biology

摘　　要：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。Objectives To evaluate the effect of an extract of fungal mycelium（EFM）on human cytomegalovirus（HCMV）transcription and to ascertain the mechanisms by which this EFM inhibits HCMV. Methods An in vitro model was created to indicate inhibition of the immediate early（IE）,early（E）,and late（L）genes of HCMV.Semi-quantitative PCR was used to compare the levels of UL122,UL123,UL44,UL57,and UL86 mRNA. Results The level of UL122 and UL123mRNA peaked 24 hafter infection,the level of UL44 and UL57mRNA peaked 96 hafter infection,and the level of UL86 mRNA peaked 120 hafter infection.The level of UL122 and UL123 mRNA decreased substantially 24 hours after cells were inoculated with the EFM and infected with HCMV or after the EFM was added to cells infected with HCMV.After the virus invaded cells,addition of 50μg/ml of the EFM resulted in a lower level of expression of UL122 and UL123mRNA than did addition of 5μg/ml of the EFM.Six h after HMCV had invaded cells,the EFM was added and cells were cultured again for 96h;the level of expression of UL44 and UL57mRNA changed little.Twentyfour h after HMCV had invaded cells,the EFM was added and cells were cultured again for 120h;the level of expression of UL86 mRNA changed little. Conclusion The EFM substantially inhibited the transcription of the IE gene of HMCV but not the E or the L gene.This is the mechanism by which the EFM inhibits HCMV.

关 键 词：真菌菌丝体提取物 巨细胞病毒 立即早期基因 早期基因 晚期基因 

分 类 号：R373[医药卫生—病原生物学] R379[医药卫生—基础医学]