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首页> 外文期刊>Journal of Cellular Physiology >Protease nexin-1: A cellular serpin down-regulated by thrombin in rat aortic smooth muscle cells.
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Protease nexin-1: A cellular serpin down-regulated by thrombin in rat aortic smooth muscle cells.

机译：蛋白酶nexin-1:细胞serpin衰减凝血酶在大鼠主动脉平滑肌细胞。

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摘要

Protease nexin-1 (PN-1), a potent inhibitor of serine proteases, is present in vascular cells and forms complexes with thrombin, plasminogen activators, and plasmin. We examined the effect of thrombin on PN-1 expression by rat aortic smooth muscle cells (RASMCs). PN-1 expression was determined by measuring protein and mRNA levels, using respectively immunoblotting and semi-quantitative reverse transcriptase polymerase chain reaction (PCR). Thrombin down-regulated PN-1 expression in a dose- and time-dependent manner. This effect was mediated via the interaction of thrombin with its receptor protease activated receptor (PAR-1) since the peptide thrombin receptor activating peptide (TRAP) reduced PN-1 expression. PN-1 secreted by smooth muscle cells remained essentially associated to cell-surface glycosaminoglycans and was released from the cell surface by heparin. A lower amount of PN-1 was released by heparin from TRAP-stimulated versus unstimulated cells and correlated with a decreased capacity to inhibit thrombin. In addition, the ability to generate peri-cellular plasmin was increased in cells with a low PN-1 expression. Pre-treatment of smooth muscle cells with cycloheximide abolished the reduction of PN-1 expression by thrombin. Furthermore, conditioned media from thrombin-treated cells reproduced the effect of thrombin, suggesting that thrombin acted via the induction of auto/paracrine mediator(s). We observed that fibroblast growth factor-2 (FGF-2)-neutralizing antibodies abolished thrombin effect whereas FGF-2 reproduced it, indicating that FGF-2 is one of the involved mediator. Together, these results indicate that (i) PN-1 modulates the activity of endogenous and exogenous serine proteases in RASMCs, (ii) thrombin down-regulates PN-1 expression and thus may increase its own activity on cells. J. Cell. Physiol. 201: 138-145, 2004. Copyright 2004 Wiley-Liss, Inc.
机译：蛋白酶nexin-1 (PN-1)的有效抑制剂丝氨酸蛋白酶,存在于血管细胞并与凝血酶形成复合物,纤溶酶原。物活化剂,胞浆素。大鼠主动脉PN-1凝血酶的表达平滑肌细胞(RASMCs)。由测量蛋白质和mRNA水平决定的,分别用免疫印迹和半定量逆转录酶聚合酶链反应(PCR)。理气PN-1剂量和表达式时间的方式。通过凝血酶和受体之间的相互作用蛋白酶激活受体(PAR-1)自肽凝血酶受体激活肽(陷阱)减少PN-1表达式。平滑肌细胞仍基本上相关的细胞表面粘多糖和从细胞表面肝素被释放。低数量的PN-1通过肝素被释放TRAP-stimulated而如果细胞与抑制能力下降有关凝血酶。peri-cellular血纤维蛋白溶酶在细胞增加一个低PN-1表达式。环己酰亚胺废除了肌肉细胞由凝血酶减少PN-1表达式。此外,条件媒体thrombin-treated细胞复制的影响凝血酶,这表明凝血酶通过行动感应自动/旁分泌的中介(年代)。观察到纤维母细胞生长因子2(FGF-2)中和抗体废除凝血酶的作用而FGF-2复制它,表明FGF-2之一调停者。(我)PN-1调节内源性的活动在RASMCs外生丝氨酸蛋白酶,(2)凝血酶下调PN-1表达式,因此可能会增加自己的细胞活动。杂志。201:138 - 145年,2004年。Wiley-Liss公司。

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