Degradation of vascular endothelial thrombomodulin by arginine- and lysine-specific cysteine proteases from Porphyromonas gingivalis.
Degradation of vascular endothelial thrombomodulin by arginine- and lysine-specific cysteine proteases from Porphyromonas gingivalis.
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DOI：
10.1902/jop.2009.090114
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发表时间：
2009-09
影响因子：
4.3
通讯作者：
M. Inomata;Y. Ishihara;T. Matsuyama;T. Imamura;I. Maruyama;T. Noguchi;K. Matsushita
中科院分区：
文献类型：
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作者：
M. Inomata;Y. Ishihara;T. Matsuyama;T. Imamura;I. Maruyama;T. Noguchi;K. Matsushita
背景：血管内皮细胞表面糖蛋白血栓调节蛋白(TM)通过调节凝血酶介导的蛋白C活化来抑制血管凝血和炎症反应。牙龈卟啉单胞菌是牙周病的主要致病菌，已在人类的血管壁和动脉粥样硬化病变中发现。已知牙龈假单胞菌来源的半胱氨酸蛋白酶(牙龈痛)可增强血管内皮细胞的炎症和凝血反应。然而，目前尚不清楚牙龈痛是否会影响血管内皮细胞TM。方法用纯化的精氨酸特异性牙龈痛(RGPS)和赖氨酸特异性牙周炎(KGP)，用免疫印迹法检测牙龈痛对重组人TM的影响。用流式细胞仪和活化蛋白C法检测牙周疼痛对血管内皮细胞表面TM的影响。采用免疫组织化学方法检测TM在牙周炎患者牙周组织微血管内皮细胞中的表达。结果RGPS和KGP对TM有体外切割作用。RGPS对内皮细胞表面TM也有降解作用。RGPS通过TM失活减少凝血酶介导的蛋白C激活。牙周炎患者牙周微血管内皮细胞TM降低。结论牙龈假单胞菌引起内皮细胞TM降解失活，可能促进血管的凝血和炎症反应。此外，TM在牙周炎患者牙周微血管内皮细胞中的表达减少，可能与牙周炎的发病机制有关。
BACKGROUND The endothelial cell surface glycoprotein thrombomodulin (TM) inhibits vascular coagulation and inflammation via regulation of thrombin-mediated activation of protein C. Porphyromonas gingivalis is the major periodontopathic bacterium and has been found in vessel walls and atherosclerotic lesions in humans. P. gingivalis-derived cysteine proteases (gingipains) are known to enhance inflammatory and coagulant responses of vascular endothelial cells. However, it has not been elucidated whether gingipains affect vascular endothelial TM. METHODS Purified arginine-specific gingipains (Rgps) and lysine-specific gingipain (Kgp) from P. gingivalis were used to investigate the effects of gingipains on recombinant human TM by immunoblot analyses. Flow cytometry and activated protein C assay were carried out to examine the effects of gingipains on vascular endothelial cell surface TM. Immunohistochemistry was performed to investigate TM expression in microvascular endothelia in gingival tissues taken from patients with periodontitis. RESULTS Rgps and Kgp cleaved TM in vitro. Endothelial cell surface TM was also degraded by Rgps. Thrombin-mediated activation of protein C was reduced by Rgps through TM inactivation. Gingival microvascular endothelial TM was reduced in patients with periodontitis. CONCLUSIONS P. gingivalis gingipains induced the degradation and inactivation of endothelial TM, which may promote vascular coagulation and inflammation. In addition, in vivo relevance was demonstrated by reduced expression of TM in gingival microvascular endothelia in patients with periodontitis, which may be involved in the pathogenesis of periodontitis.