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，对免疫系统中的许多发育过程至关重要。钙调磷酸酶在免疫应答中的作用被钙调磷酸酶抑制剂如环孢菌素A和FK 506所强调。我们以前已经证明，促炎细胞因子，IL-16和TNF-α诱导人牙龈成纤维细胞释放PGE_2。本研究旨在探讨钙调磷酸酶抑制剂对牙龈成纤维细胞PGE_2合成的影响。结果表明，环孢素A可诱导前列腺素E_2的释放，而FK 506则可诱导COX-2的表达。当用IL-16处理人牙龈成纤维细胞时，这种诱导更加突出。环孢菌素A以剂量和时间依赖的方式诱导PGE 2的释放，在10 μM作用24 h时达到最大值。环孢菌素A和FK 506部分增加IL-18介导的PGE_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PGE_2释放。RT-PCR分析表明，环孢菌素A增加环氧合酶-2 mRNA的表达。与单独用IL-1β相比，用环孢素A和IL-16同时处理导致环氧合酶-2 mRNA表达增加。为了确认RT-PCR结果，通过定量真实的时间RT-PCR测量环加氧酶转录物水平。这些真实的实时RT-PCR数据支持RT-PCR结果，证实环孢菌素A上调环氧合酶-2 mRNA。环孢霉素A可能通过诱导人牙龈成纤维细胞环氧化酶2介导的PGE_2生成而发挥炎症介质的作用。综上所述，我们的数据表明，环孢菌素A诱导环氧合酶-2和PGE_2的合成增强，它们可能参与环孢菌素A诱导的牙龈增生。

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.