Calcineurin is involved in inflammatory responses in human gingival fibroblasts

钙调神经磷酸酶是一种钙激活的丝氨酸/苏氨酸磷酸酶，在免疫系统的许多发育过程中起关键作用。钙调神经磷酸酶抑制剂，如环孢素A和FK506，强调了钙调神经磷酸酶在免疫反应中的作用。我们先前已经证明，促炎细胞因子、IL-16和TNF-α可诱导人牙龈成纤维细胞释放前列腺素E_2。在这项研究中，我们研究了钙调神经磷酸酶抑制剂是否影响人牙龈成纤维细胞前列腺素E_2的合成。我们分析了环孢素A和FK506对前列腺素E_2释放和环氧合酶-2 mRNA表达的影响。环孢素A诱导环氧合酶-2表达以及前列腺素E_2释放。当人牙龈成纤维细胞被IL-16处理时，这种诱导作用更加明显。环孢菌素A以剂量和时间依赖的方式诱导前列腺素E_2释放，并在10μM作用24小时达到最大。环孢素A和FK506可部分增加IL-18介导的前列腺素E_2释放。此外，在IL-1β预处理的牙龈成纤维细胞中，环孢菌素A可促进缓激肽刺激的PGE2释放。RT-PCR分析显示，环孢素A可上调环氧合酶-2mRNA的表达。与单独使用IL-1β相比，联合应用环孢素A和IL-16可增加环氧合酶-2mRNA的表达。为了验证RT-PCR结果，用实时定量RT-PCR方法检测环氧合酶的转录水平。这些实时RT-PCR数据支持RT-PCR结果，证实环氧合酶-2mRNA被环孢素A上调，环孢素A可能作为炎症介质，可能通过诱导环氧合酶-2介导的PGE2的形成而在人牙龈成纤维细胞中发挥作用。综上所述，我们的数据提示环氧合酶-2和前列腺素E_2合成的诱导被环孢素A增强，它们可能参与环孢素A诱导的牙龈过度生长。

Calcineurin is a calcium-activated serine/threonine phosphatase critical to a number of developmental processes in the immune systems. The role of Calcineurin in the immune response is underscored by the calcineurin inhibitors, such as cyclosporin A and FK506. We have previously demonstrated that pro-inflammatory cytokines, IL-16 and TNF-a induced PGE_2 release in human gingival fibroblasts. In this research project, we investigated whether Calcineurin inhibitors affect PGE_2 synthesis in human gingival fibroblasts. We analyzed the effects of cyclosporin A and FK506 on the PGE_2 release and the expression of cyclooxygenase-2 mRNA.Cyclosporin A induced cyclooxygenase-2 expression as well as prostaglandin E_2 release. This induction was more prominent when human gingival fibroblasts were treated with IL-16. Cyclosprin A induced PGE2 release in dose-and time-dependent manner, and reached a maximum at 10 μM for 24h. Cyclosporin A and FK506 partially increased IL-18-mediated PGE_2 release. Moreover, in the gingival fibroblasts pretreatment with IL-1β, cyclosporin A augmented bradykinin-stimulated PGE_2 release. RT-PCR analysis revealed that cyclosporin A increased the expression of cyclooxygenase-2 mRNA. The simultaneous treatment with cyclosporin A and IL-16 resulted in increased the expression of cyclooxygenase-2 mRNA compared to cells with IL-1β alone. To confirm the RT-PCR results, cyclooxygenase transcript levels were measured by quantitative real time RT-PCR. These real time RT-PCR data supported the RT-PCR results, confirming that cyclooxygenase-2 mRNA was up-regulated by cyclosporin A. Cyclosporin A could acts as an inflammatory mediator, likely by the induction of cyclooxygenase-2-mediated PGE_2 formation in human gingival fibroblasts. Taken together, our data suggest that induction of cyclooxygenase-2 and PGE_2 synthesis are enhanced by cyclosporine A where they may participate in cyclosporin A-induced gingival overgrowth.