Herpes simplex virus (HSV) infection induces a rapid and transient increase in intracellular calcium concentration ([Ca²⁺]_i), which plays a critical role in facilitating viral entry. T-type calcium channel blockers and EGTA, a chelate of extracellular Ca²⁺, suppress HSV-2 infection. But the cellular mechanisms mediating HSV infection-activated Ca²⁺ signaling have not been completely defined. In this study we investigated whether the TRPV4 channel was involved in HSV-2 infection in human vaginal epithelial cells. We showed that the TRPV4 channel was expressed in human vaginal epithelial cells (VK2/E6E7). Using distinct pharmacological tools, we demonstrated that activation of the TRPV4 channel induced Ca²⁺ influx, and the TRPV4 channel worked as a Ca²⁺-permeable channel in VK2/E6E7 cells. We detected a direct interaction between the TRPV4 channel protein and HSV-2 glycoprotein D in the plasma membrane of VK2/E6E7 cells and the vaginal tissues of HSV-2–infected mice as well as in phallic biopsies from genital herpes patients. Pretreatment with specific TRPV4 channel inhibitors, GSK2193874 (1−4 μM) and HC067047 (100 nM), or gene silence of the TRPV4 channel not only suppressed HSV-2 infectivity but also reduced HSV-2-induced cytokine and chemokine generation in VK2/E6E7 cells by blocking Ca²⁺ influx through TRPV4 channel. These results reveal that the TRPV4 channel works as a Ca²⁺-permeable channel to facilitate HSV-2 infection in host epithelial cells and suggest that the design and development of novel TRPV4 channel inhibitors may help to treat HSV-2 infections.

单纯疱疹病毒 (HSV) 感染会诱导细胞内钙浓度 ([Ca ²⁺ ] _i ) 快速短暂增加，这在促进病毒进入方面发挥着关键作用。 T 型钙通道阻滞剂和 EGTA（细胞外 Ca ²⁺螯合物）可抑制 HSV-2 感染。但介导 HSV 感染激活的 Ca ²⁺信号传导的细胞机制尚未完全明确。在本研究中，我们研究了 TRPV4 通道是否参与人阴道上皮细胞的 HSV-2 感染。我们发现 TRPV4 通道在人阴道上皮细胞 (VK2/E6E7) 中表达。使用不同的药理学工具，我们证明TRPV4通道的激活诱导Ca ²⁺流入，并且TRPV4通道在VK2/E6E7细胞中充当Ca ²⁺通透通道。我们在 VK2/E6E7 细胞质膜、HSV-2 感染小鼠的阴道组织以及生殖器疱疹患者的阴茎活检中检测到 TRPV4 通道蛋白和 HSV-2 糖蛋白 D 之间的直接相互作用。用特定的 TRPV4 通道抑制剂 GSK2193874 (1−4 μM) 和 HC067047 (100 nM) 进行预处理，或 TRPV4 通道的基因沉默不仅抑制了 HSV-2 的感染性，而且还减少了 VK2/ 中 HSV-2 诱导的细胞因子和趋化因子的产生。 E6E7 细胞通过 TRPV4 通道阻断 Ca ²⁺流入。这些结果表明，TRPV4通道作为Ca ²⁺渗透通道，促进宿主上皮细胞中的HSV-2感染，并表明新型TRPV4通道抑制剂的设计和开发可能有助于治疗HSV-2感染。