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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。
This article mainly discusses the role of PKC and PKA signaling in the regulation of VEGF expression in cultured human dental follicle cells. The fourth generation human dental follicle cells with good growth condition were selected and tested by Real-time PCR and Western blot for the detection of PKC agonist (PMA), PKC non-specific inhibitor (G 6983), PKC-α and γ specific inhibitors HBDDE, PKC-β specific inhibitor (LY333531), PKA agonist (dbcAMP) and inhibitor (KT5720) on the expression of VEGF mRNA and protein in cultured human dental follicle cells. The results showed that the expression levels of VEGF mRNA and protein in PMA group and PMA + HBDDE group were significantly higher than those in control group (P <0.05), while the expression of VEGF mRNA and protein in PMA + G 6983 group and PMA + LY333531 group There was no significant difference between the expression levels and the control group (P> 0.05). The expression level of VEGF mRNA and protein in dbcAMP group was significantly higher than that in control group (P <0.05), while there was no significant difference between dbcAMP + KT5720 group and control group (P> 0.05 ). This indicates that both PKC and PKA signaling pathways are involved in the regulation of VEGF expression in cultured human dental follicle cells, of which the PKC signaling pathway involved in the regulation of PKC-β.