人干扰素结合蛋白35在人巨细胞病毒感染过程中的表达及意义

摘要

目的 探讨人干扰素结合蛋白35(interferon-induced protein 35,IFP35)在人巨细胞病毒感染过程中的表达及意义.方法 依据是否感染人巨细胞病毒将血细胞分为感染组和未感染组;根据人包皮成纤维细胞转染siRNA和阴性siRNA分为基因敲减组和阴性对照组;采用SYBRGreen荧光定量聚合酶链式反应(polymerase chain reaction,PCR)和蛋白质印迹法检测不同分组中IFP35的表达量;用人巨细胞病毒感染人包皮成纤维细胞,采用SYBRGreen荧光定量PCR和蛋白质印迹法检测不同感染复数的人巨细胞病毒感染人包皮成纤维细胞表达IFP35的差异以及相同感染复数的人包皮成纤维细胞感染人巨细胞病毒0、12、24、48、96小时后IFP35表达的差异;敲减IFP35基因后,检测人巨细胞病毒量的变化情况.结果 感染组血细胞中IFP35表达量显著高于未感染组(P<0.05).感染后细胞中IFP35的表达量随人巨细胞病毒浓度的增加而增加,与经感染复数为0的人巨细胞病毒感染后的细胞比较,经感染复数为0.01、0.1及1.0的人巨细胞病毒感染后细胞中IFP35的表达量均显著升高(P<0.05).IFP35的表达量随人巨细胞病毒感染时间的延长而增加,且与0小时相比,其余时间其表达量均显著增加(P<0.05).基因敲减组人包皮成纤维细胞中IFP35的表达水平显著低于阴性对照组(P<0.05).敲减IFP35基因的人巨细胞病毒DNA基因拷贝数较未敲减IFP35基因的人巨细胞病毒显著增加(P<0.05).结论 IFP35的表达量与人巨细胞病毒感染的浓度和时间均呈正相关.IFP35基因的表达可能在一定程度上抑制人巨细胞病毒的感染.%Objective To investigate the expression and significance of interferon-induced protein 35 (IFP35) in human cytomegalovirus infection. Method Cells infected with human cytomegalovirus would be divided into infected group and uninfected group, and human foreskin fibroblast cells were divided into gene knockout group and negative control group according to transfection of siRNA and negative siRNA. The difference of IFP35 expression in human foreskin fibroblast cells infected with different multiplicity of infection values and human foreskin fibroblast cells infected with human cytomegalovirus 0, 12, 24, 48 and 96 hours after the same multiplicity of infection value were detected by qPCR and Western blotting. Through the knockdown experiment. After knocking down the IFP35 gene, detected the amount of human cytomegalovirus virus. Result The expression of IFP35 in infected group was significantly higher than that in uninfected group (P < 0.05). With the increase of human cytomegalovirus concentration, the expression level of IFP35 cells infected increased. Compared with multiplicity of infection at 0, the expression levels of IFP35 in cells with multiplicity of infection at 0.01, 0.1 and 1.0 significantly increased (P < 0.05). The expression level of IFP35 increased with the time of human cytomegalovirus infection. Compared with 0 hour, the expression levels of IFP35 after the rest of the time significantly increased (P < 0.05). After IFP35 gene knockdown, the expression level of IFP35 of human foreskin fibroblasts in gene knockout group was significantly lower than that of negative control group (P < 0.05) The number of human cytomegalovirus DNA gene copies with IFP35 gene knockdown was significantly higher than that without IFP35 gene knockdown (P < 0.05). Conclusion The expression level of IFP35 is positively correlated with human cytomegalovirus infection concentration and time. Maybe the expression of IFP35 gene inhibits human cytomegalovirus infection to a certain extent.