摘要

苯乙烯吡喃衍生物在HSV-1感染Vero细胞中的差异基因表达作用

作者(年代):Noor ZarinaAbd Wahab, Nazlina Ibrahim,Halimah Abdullah Sani和AzimahtolHawariah Lope Pihie

本研究旨在鉴定和表征HSV-1感染宿主和苯乙烯吡咯酮衍生物(SPD)处理宿主过程中特定细胞基因的表达。SPD对HSV-1具有不同作用方式的抗病毒活性。SPD在感染后2 ~ 4小时加入，可有效抑制细胞死亡。只有当治疗延迟到感染后5和6小时时，才观察到细胞死亡。这种作用模式的积极作用表明SPD能够在治疗模式[S+V]+SPD的两种有效浓度(1.563x10-7µM和7.813x10-8µM)下治疗HSV-1感染细胞。用差异基因表达法(DEG)测定和分离在SPD处理和未处理的HSV-1感染Vero细胞中差异表达的基因。DEG分析结果显示，共有177个基因表达差异，其中89个候选cdna被诱导表达，88个候选cdna被抑制表达。通过生物信息学分析，将所有基因的核苷酸序列与Genbank数据库进行比较。从DEG分析中筛选出8个标记，采用Real- Time PCR对其表达进行了验证。Real-Time PCR结果显示markersÂÃ、Â的表达谱呈100%的相关性。 The cDNA markers that were induced in their expression include mitogen-activated protein kinase, tapasin, carboxypeptidase M, RAB member RAS oncogen family, p53 and G protein-coupled receptor.We found that SPD induced apoptosis, as measured by oncogene family gene expression and caspase activation. The apoptosis mediated by SPD in infected cells was associated with the activation of p53 and Bcl-2 protein via intrinsic pathway. SPD also exerts its anti-HSV-1 properties by activating an extrinsic pathway via caspase-8 activation in infected cells. From this study, the understanding on how SPD acted upon HSV-1 infected host cells during the infection process was proposed. In this study it was shown that SPD has a potential in controlling HSV-1 infection selectively without disturbing non-infected cells.