Koziak K, Bojakowska M, Robson S C, Bojakowski K, Soin J, Csizmadia E, Religa P, Gaciong Z, Kaczmarek E
Department of General and Nutritional Biochemistry, The Medical University of Warsaw, Warsaw, Poland.
J Thromb Haemost. 2008 Jul;6(7):1191-7. doi: 10.1111/j.1538-7836.2008.03019.x. Epub 2008 Jul 1.
Growing evidence implicates the involvement of extracellular nucleotides in the regulation of platelet, leukocyte, endothelial cell (EC) and vascular smooth muscle cell (VSMC) phenotype and function. Within the quiescent vasculature, extracellular nucleotides are rapidly hydrolyzed by CD39, the dominant endothelial nucleoside triphosphate diphosphohydrolase (NTPDase-1). However, vascular CD39/NTPDase-1 activity is lost in EC activated by oxidative stress or proinflammatory mediators, and upon denudation of the endothelium following balloon injury. The consequent increase in extracellular nucleotide concentrations triggers signaling events leading to prothrombotic responses and increased VSMC proliferation.
To investigate the effect of overexpressed CD39/NTPDase-1 in injured aorta.
Using adenoviral-mediated gene transfer we expressed CD39/NTPDase-1 in mechanically denudated rat aortas. We measured intima formation by morphometry and VSMC proliferation by the [(3)H]-thymidine incorporation assay.
Targeted expression of CD39 in injured vessels increased NTPDase activity (from 2.91 +/- 0.31 to 22.07 +/- 6.7 nmols Pi mg(-1) protein, 4 days after exposure to the adenovirus) and prevented the formation of neointima. The thickness of the intimal layer in injured aortas exposed to Ad-CD39 was 26.2 +/- 3.9 microm vs. 51.8 +/- 6.1 microm and 64.4 +/- 22.2 microm (P < 0.001) in vessels treated with Ad-beta-gal and saline, respectively. Moreover, targeted expression of CD39/NTPDase-1 caused a 70% (P < 0.01) decrease in proliferation of VSMC isolated from transduced rat aortas as compared with VSMC derived from control vessels.
The presented data suggest that increasing CD39/NTPDase-1 activity in VSMC could represent a novel therapeutic approach for the prevention of stenosis associated with angioplasty and other occlusive vascular diseases.
越来越多的证据表明，细胞外核苷酸参与血小板、白细胞、内皮细胞（EC）和血管平滑肌细胞（VSMC）表型及功能的调节。在静止的脉管系统中，细胞外核苷酸被CD39迅速水解，CD39是主要的内皮核苷三磷酸二磷酸水解酶（NTPDase-1）。然而，在受到氧化应激或促炎介质激活的内皮细胞中，以及在球囊损伤后内皮剥脱时，血管CD39/NTPDase-1活性丧失。细胞外核苷酸浓度的相应增加引发信号转导事件，导致促血栓形成反应和VSMC增殖增加。
研究过表达CD39/NTPDase-1对损伤主动脉的影响。
使用腺病毒介导的基因转移，在机械剥脱的大鼠主动脉中表达CD39/NTPDase-1。我们通过形态计量学测量内膜形成，并通过[（3）H]-胸腺嘧啶核苷掺入试验测量VSMC增殖。
在损伤血管中靶向表达CD39可增加NTPDase活性（在暴露于腺病毒4天后，从2.91±0.31增加到22.07±6.7 nmol Pi mg（-1）蛋白），并防止新生内膜形成。暴露于Ad-CD39的损伤主动脉内膜层厚度为26.2±3.9微米，而用Ad-β-半乳糖苷酶和生理盐水处理的血管中分别为51.8±6.1微米和64.4±22.2微米（P<0.001）。此外，与来自对照血管的VSMC相比，靶向表达CD39/NTPDase-1使从转导的大鼠主动脉分离的VSMC增殖减少70%（P<0.01）。
所提供的数据表明，增加VSMC中CD39/NTPDase-1活性可能是预防与血管成形术和其他闭塞性血管疾病相关的狭窄的一种新的治疗方法。
