The nuclear factor of activated T-cell (NFAT) signaling pathway is involved in angiogenesis following initiation by vascular endothelial growth factor (VEGF). A number of angiogenic genes have been associated with calcineurin in the NFAT pathway, forming a calcineurin-NFAT pathway. This study aims to investigate the involvement of four angiogenic genes within the calcineurin-NFAT pathway in the endothelial-like differentiation of stem cells from human exfoliated deciduous teeth (SHED) cultured on a human amniotic membrane (HAM) induced by VEGF. SHED were induced with VEGF for 24 h, then cultured on the stromal side of HAM. The cells were then further induced with VEGF until days 1 and 14. To understand the role of calcineurin, its potent inhibitor, cyclosporin A (CsA), was added into the culture. Results from SEM and H&E analyses showed SHED grew on HAM surface. Gene expression study of Cox-2 showed a drastically reduced expression with CsA treatment indicating Cox-2 involvement in the calcineurin-NFAT pathway. Meanwhile, IL-8 was probably controlled by another pathway as it showed no CsA inhibition. In contrast, high expression of ICAM-1 and RCAN1.4 by VEGF and CsA implied that these genes were not controlled by the calcineurin-NFAT-dependent pathway. In conclusion, the results of this study suggest the involvement of Cox-2 in the calcineurin-NFAT-dependent pathway while RCAN1.4 was controlled by NFAT molecule in endothelial-like differentiation of SHED cultured on HAM with VEGF induction.

活化 T 细胞 (NFAT) 信号通路的核因子参与血管内皮生长因子 (VEGF) 启动后的血管生成。许多血管生成基因与 NFAT 通路中的钙调神经磷酸酶相关，形成钙调神经磷酸酶-NFAT 通路。本研究旨在研究钙调神经磷酸酶-NFAT 通路中的四种血管生成基因在 VEGF 诱导的人羊膜 (HAM) 上培养的人脱落乳牙 (SHED) 干细胞的内皮样分化中的参与。SHED 用 VEGF 诱导 24 h，然后在 HAM 的基质侧培养。然后用 VEGF 进一步诱导细胞，直到第 1 天和第 14 天。为了了解钙调神经磷酸酶的作用，将其强效抑制剂环孢菌素 A (CsA) 添加到培养物中。SEM 和 H& 的结果 E 分析表明 SHED 在 HAM 表面生长。基因表达研究Cox-2在 CsA 处理下表现出显着降低的表达，表明Cox-2参与钙调神经磷酸酶-NFAT 途径。同时，IL-8可能受另一种途径控制，因为它没有表现出 CsA 抑制。相比之下，VEGF 和 CsA对ICAM-1和RCAN1.4的高表达暗示这些基因不受钙调神经磷酸酶-NFAT依赖性途径的控制。总之，这项研究的结果表明Cox-2参与钙调神经磷酸酶-NFAT依赖性途径，而RCAN1.4受 NFAT 分子控制，在 HAM 上培养的SHED内皮样分化中，VEGF 诱导。