Objective To investigate the effect of human herpes simplex virus type Ⅱ (HSV-2) on intercellular tight junction of genital epithelial cells and the chemotaxis of monocytes.Methods The in vitro genital epithelial tight junction monolayer model based on HEC-1-A Millicell culture was developed.After infected with low dose HSV-2 (MOI=0.1),the cell viability was tested and the transepithelial electrical resistance was measured at different time points to evaluate intercellular tight junction of HEC-1-A cells.The expression of ZO-1 protein was detected with Western blot and RT-PCR.The in vitro monocyte chemotaxis model was used through inducing U937 cell to express oligopeptide chemotaxis receptor,and monocyte chemotaxis was detected after HSV-2 infected HEC-1-A culture supernatant was added.Results The transepithelial electrical resistance of HEC-1-A cells was reduced by HSV-2 infection (MOI=0.1) gradually,especially in the middle and late period (P＜0.01),and the cell viability had no significant changes (above 80％).HSV-2 infection down-regulated the protein expression of ZO-1 in HEC-1-A cells,but had no effect on E-cadherin and Occuldin expression (P ＞0.05).The supernatant from HSV-2 infected HEC-1-A cells culture strongly induced monocyte chemotaxis (P＜0.01).Conclusion Low dose HSV-2 infection can reduce transepithelial electrical resistance of HEC-1-A cells and damage cell tight junction,which may be related to down-regulation of ZO-1 protein;and also induce the chemotaxis of monocytes by release of chemotactic factors.%目的 探讨人类单纯疱疹病毒Ⅱ型(HSV-2)感染对生殖道上皮细胞间紧密连接的影响及对单核细胞的趋化作用.方法 通过以人子宫内膜腺癌细胞(HEC-1-A)小室培养模型为基础建立生殖道体外单层细胞致密层模型,低剂量HSV-2感染后(MOI=0.1),检测体外细胞活力;通过检测跨膜电阻判断细胞间紧密连接程度的变化;采用Western blot、荧光定量PCR法观察细胞紧密连接蛋白ZO-1、E-cadherin和Occuldin的表达情况;通过单核细胞趋化反应实验来检测感染HSV-2的HEC-1-A细胞培养上清液的趋化效应.结果 与阴性对照组(Mock组)相比,HSV-2(MOI=0.1)感染使HEC-1-A单层细胞致密层跨膜电阻逐渐降低,且在感染中后期下降明显(P＜0.01),而细胞活力无显著变化(均＞80％);随着HSV-2病毒感染,ZO-1蛋白表达水平逐渐降低,在感染60h,该蛋白表达水平最低,而其mRNA表达水平无显著变化(P＞0.05),E-cadherin和Occuldin均无显著变化(均P＞0.05);HSV-2感染HEC-1-A后的细胞上清液可诱导单核细胞产生明显的趋化反应(P＜0.01).结论 低剂量HSV-2持续感染可降低HEC-1-A单层细胞致密层跨膜电阻,破坏细胞间紧密连接,很可能与下调ZO-1表达水平相关;感染后的细胞上清液可产生某种特殊的趋化分子介导免疫细胞向感染区域迁移浸润.
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