该文主要探讨PKC、PKA信号通路在调控体外培养人牙囊细胞VEGF表达中的作用。选取生长状态良好的第4代人牙囊细胞,采用Real-time PCR和Western blot分别检测PKC激动剂(PMA)、PKC非特异性抑制剂(G 6983)、PKC-α和γ特异性抑制剂(HBDDE)、PKC-β特异性抑制剂(LY333531)、PKA激动剂(dbcAMP)和抑制剂(KT5720)对体外培养人牙囊细胞VEGF mRNA和蛋白表达的影响。结果显示,PMA组和PMA+HBDDE组VEGF mRNA和蛋白的表达水平明显高于对照组,差异有统计学意义(P〈0.05);而PMA+G 6983组和PMA+LY333531组VEGF mRNA和蛋白的表达水平与对照组之间无明显差异(P〉0.05)。dbcAMP组VEGF mRNA和蛋白的表达水平明显高于对照组,差异有统计学意义(P〈0.05);而dbcAMP+KT5720组VEGF mRNA和蛋白的表达水平与对照组之间无明显差异(P〉0.05)。这表明,PKC、PKA信号通路均参与了体外培养人牙囊细胞VEGF表达的调控,其中PKC信号通路中参与调控的亚型是PKC-β。
The aim of this paper is to study whether PKC and PKA signaling pathways are involved in regulation of expression of VEGF in cultured human dental follicle cells (HDFC). HDFC in good status were incubated with PMA (PKC activator), PMA+G66983 (PKC non-specific inhibitor), PMA+HBDDE (PKC-α and y specific inhibitor), PMA+LY333531 (PKC-β specific inhibitor), dbcAMP (PKA activator), dbcAMP+KT5720 (PKA inhibitor) for 2 h, respectively. Real-time PCR and Western blot were used to detect the gene and protein expression of VEGF in these groups respectively. The results showed that the gene and protein expression of VEGF in the group with PMA alone or the group with PMA+HBDDE were significantly higher than that of the control (P〈0.05). There was no significant difference in the gene and protein expression of VEGF between the group with PMA+G66983 (or the group with PMA+LY333531) and the control (P〉0.05). dbeAMP can up-regulate the gene and protein expression of VEGF in cultured HDFC, while this effect can be inhibited by KT5720. In conclusion, these findings suggest that the expression of VEGF in cultured HDFC can be regulated via PKC and PKA signaling pathways. In PKC signal- ing pathway, PKC-β may be the isoform that regulates VEGF expression in cultured HDFC.