Inductive effect of blood coagulation factors from endothelial cells by Porphyromonas gingivalis fimbriae

为了研究外源性凝血途径，必须将因子VII激活至VIIa。已知胰蛋白酶可以激活因子VII到VIIa。本文表明，在人冠状动脉内皮细胞（HCAEC）培养中，牙龈卟啉单胞菌菌毛刺激胰蛋白酶原mRNA的表达及其蛋白的产生。将单层HCAEC与不同剂量的菌毛孵育。采用RT-PCR和western blot检测和分析胰蛋白酶原在HCAEC中的表达。菌毛刺激胰蛋白酶原mRNA的表达及其蛋白的产生呈剂量依赖性。利用放射性标记的纤维结合试验证实了纤维与HACEC的结合。此外，抗菌膜抗体（EF3-11）可抑制菌膜诱导的胰蛋白酶原在HCAEC中的表达。最后，PKC抑制剂H7抑制了菌毛对胰蛋白酶原产生的诱导作用。这些结果表明，菌毛结合HCAEC，随后导致异常凝血因子的刺激。这些结果提示牙周病细菌在人心脏血管系统血液凝固异常中的可能作用。

To Investigate extrinsic blood coagulation pathway, factor VII must be activated to VIIa. Trypsin is known to activate factor VII to VIIa. Here I show that Porphyromonas gingivalis fimbriae stimulate trypsinogen mRNA expression and its protein production in human coronary artery endothelial cells (HCAEC) culture. The monolayer of HCAEC were incubated with or with out various doses of the fimbriae. RT-PCR and western blot analysis were perfomed ti detect and analyze trypsinogen expression in HCAEC. The fimbriae stimulated the expression of trypsinogen mRNA and production of its protein in a fimbriae dose dependent manner. The fimbriae binding to HACEC was specifically evidenced by a binding assay involving radio labeled fimbriae. Additionally, anti-fimbriae antibody (EF3-11) inhibited the fombriae-induces expression of trypsinogen in HCAEC. Finally, inductive effect of trypsinogen production by the fimbriae in HCAEC were inhibited by H7 an inhibitor of PKC. These results demonstrated that fimbriae bind HCAEC, which subsequently lead to stimukate of abnormal blood coagulation factor. These results thus suggest that the possible roles of the periodontopathesic bacteria in abnormal blood coagulation in human heart vascular system.