Expression and activity of thrombomodulin in human gingival epithelium: in vivo and in vitro studies.
Expression and activity of thrombomodulin in human gingival epithelium: in vivo and in vitro studies.
复制标题
人牙龈上皮中血栓调节蛋白的表达和活性：体内和体外研究。
DOI：
10.1034/j.1600-0765.2000.035003146.x
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发表时间：
2000
影响因子：
3.5
通讯作者：
T. Sueda
中科院分区：
文献类型：
--
作者：
T. Matsuyama;Y. Izumi;K. Shibatate;Y. Yotsumoto;H. Obama;M. Uemura;I. Maruyama;T. Sueda
表皮角质形成细胞血栓调节蛋白（TM）已被证明除了表皮分化的作用外，还可调节皮肤损伤部位的凝血酶。TM是内皮细胞膜的主要抗凝蛋白聚糖，是凝血酶受体，作为蛋白C活化的辅因子。凝血酶对牙周炎有促炎作用。然而，很少有人知道TM在牙周炎牙龈组织。我们用免疫组织化学方法检测TM在牙周炎患者牙龈上皮中的表达。在体外培养条件下，我们通过激光共聚焦显微镜观察不同Ca2+浓度下TM的表达，检测TM mRNA的表达，并检测TM辅因子的活性。此外，我们还用酶联免疫吸附法测定了11例成人重度牙周炎患者龈沟液（GCF）中TM浓度。免疫反应性TM存在于牙龈上皮和结合上皮中，并且与健康牙龈上皮相比，在发炎的牙龈上皮中减少。超微结构上，TM，包括微绒毛，观察细胞膜上。TM定位在0.09 mM Ca2+培养的细胞中的不同，暴露于1.2 mM Ca2+的细胞。北方分析显示TM mRNA在牙龈角质形成细胞中的表达高于人脐静脉内皮细胞（HUVEC）。牙龈角质形成细胞也促进凝血酶激活蛋白C，但不如HUVEC强烈。在探诊出血的患者部位，GCF中的TM显著升高（p < 0.001，Student's t检验）。牙龈上皮中的TM可以调节牙周病凝血和炎症部位的凝血酶活性，尽管炎症可能会损害凝血酶的这种调节。
Epidermal keratinocytes thrombomodulin (TM) has been shown to regulate thrombin at sites of cutaneous injury in addition to a role for epidermal differentiation. TM, a major anticoagulant proteoglycan of the endothelial cell membrane, is a thrombin receptor that acts as a co-factor for protein C activation. Thrombin has pro-inflammatory effects for periodontitis. However, little is known about TM in gingival tissue with periodontitis. We used immunohistochemistry to examine expression of TM in gingival epithelium from patients with periodontitis. In vitro, we observed TM expression at varying Ca2+ concentrations by confocal laser scanning microscopy, examined the expression of TM mRNA and tested TM co-factor activity. Furthermore, we measured TM concentration in gingival crevicular fluid (GCF) from 11 severe adult cases of periodontitis using enzyme-linked immunosorbent assay. Immunoreactive TM was present in gingival epithelium and junctional epithelium, and was reduced in inflamed gingival epithelium compared to healthy gingival epithelium. Ultrastructurally, TM, including microvilli, was observed on the cell membrane. TM localization in cells cultured in 0.09 mM Ca2+ differed from that in cells exposed to 1.2 mM Ca2+. Northern analysis demonstrated TM mRNA in gingival keratinocytes more than in human umbilical vein endothelial cells (HUVEC). Gingival keratinocytes also facilitated protein C activation by thrombin, although less strongly than HUVEC. TM in GCF at sites with bleeding on probing in patients was significantly elevated (p < 0.001, Student's t-test). TM in gingival epithelium may regulate thrombin activity at sites of coagulation and inflammation with periodontal disease, although inflammation may impair this regulation of thrombin.