Saito Hiroki, Kitamoto Masayuki, Kato Kozue, Liu Ning, Kitamura Hisayo, Uemura Kazuhide, Nogaki Fumiaki, Takeda Toshiya, Mori Noriko, Ono Takahiko
Division of Molecular Medicine, University of Shizuoka School of Pharmaceutical Sciences, Shizuoka, Japan.
Perit Dial Int. 2009 May-Jun;29(3):340-51.
Fibrin deposition on the peritoneum has been frequently observed in peritoneal fibrosis induced by long-term peritoneal dialysis. The present study was conducted to clarify the contribution of factor Xa through tissue factor and factor V expression in peritoneal fibrosis.
Wistar rats were intraperitoneally injected with chlorhexidine gluconate (CG) every day. For the interventional study, the factor Xa inhibitor fondaparinux was subcutaneously administered. After 28 days of CG injection, peritoneal specimens were examined by immunohistochemical analyses and in situ hybridization.
The peritoneal submesothelial compact zone was observed to be markedly thicker in the CG-injected groups than in the normal group, and that thickness was dose dependent. Immunohistochemical study revealed massive fibrin, fibronectin, and type IV collagen depositions in the CG-injected groups, which was markedly higher than that in the normal group. Macrophage infiltration and staining for tissue factor, factor V, factor X, and protease-activated receptor-2 were intense in the CG-injected groups and negative/trace in the normal group. Tissue factor and factor V mRNAs were abundant in cells in the thickened peritoneum. A double-labeling experiment revealed that tissue factor was observed mainly in macrophages, and factor V was abundantly distributed in the fibrotic tissue together with macrophages. Fondaparinux treatment decreased the thickness of submesothelial fibrotic tissue, and size and number of CD31-positive vessels.
These results suggest that expression of tissue factor and factor V in infiltrated macrophages, together with factor X deposition, may progress angiogenesis and accumulation of extracellular matrix components, partly via profibrotic and procoagulant mechanisms in the peritoneum after inflammatory stimulation.
在长期腹膜透析所致的腹膜纤维化中，经常观察到纤维蛋白在腹膜上的沉积。本研究旨在阐明凝血因子Xa通过组织因子和凝血因子V的表达在腹膜纤维化中的作用。
每天给Wistar大鼠腹腔注射葡萄糖酸氯己定（CG）。在干预研究中，皮下注射凝血因子Xa抑制剂磺达肝癸钠。CG注射28天后，通过免疫组织化学分析和原位杂交检查腹膜标本。
观察到CG注射组的腹膜间皮下致密带明显比正常组厚，且该厚度呈剂量依赖性。免疫组织化学研究显示，CG注射组有大量纤维蛋白、纤连蛋白和IV型胶原沉积，明显高于正常组。CG注射组巨噬细胞浸润以及组织因子、凝血因子V、凝血因子X和蛋白酶激活受体-2的染色强烈，而正常组为阴性/微量。增厚腹膜中的细胞富含组织因子和凝血因子V的mRNA。双重标记实验显示，组织因子主要在巨噬细胞中观察到，凝血因子V与巨噬细胞一起大量分布在纤维化组织中。磺达肝癸钠治疗可降低间皮下纤维化组织的厚度以及CD31阳性血管的大小和数量。
这些结果表明，浸润巨噬细胞中组织因子和凝血因子V的表达，连同凝血因子X的沉积，可能通过炎症刺激后腹膜中的促纤维化和促凝血机制，促进血管生成和细胞外基质成分的积累。
