Antithrombin reduces shedding of the endothelial glycocalyx following ischaemia/reperfusion
Antithrombin reduces shedding of the endothelial glycocalyx following ischaemia/reperfusion
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DOI：
10.1093/cvr/cvp097
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发表时间：
2009-07-15
影响因子：
10.8
通讯作者：
Becker, Bernhard F.
中科院分区：
文献类型：
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作者：
Chappell, Daniel;Jacob, Matthias;Becker, Bernhard F.
抗凝血酶是凝血系统的重要抑制剂，另外对内皮细胞发挥特异性抗炎作用。健康的血管内皮被内皮糖萼包被，其减少增加毛细血管通透性，例如在缺血后。已知抗凝血酶可渗入糖萼，与糖胺聚糖结合，并在应用肿瘤坏死因子-α后保护糖萼。用Krebs-Henseleit缓冲液（KHB）灌流离体豚鼠心脏，观察抗凝血酶对缺血/再灌注后糖萼的影响。在诱导20分钟缺血（37 ℃）之前，应用抗凝血酶以达到生理水平（1 U/mL）。用KHB或KHB加2 g%羟乙基淀粉（130 kDa）以恒定流量（基线灌注压70 cmH（2）O）再灌注心脏20 min。通过测量心外膜表面上的渗出液形成直接评估冠状动脉净液体过滤。通过ELISA定量缺血后冠状动脉释放的多配体蛋白聚糖-1和硫酸乙酰肝素。将心脏灌注固定以通过电子显微镜观察糖萼。缺血/再灌注导致糖萼降解，冠状动脉灌注压升高，血管通透性增加。与未经治疗的心脏相比，抗凝血酶显著减少缺血后糖萼脱落、冠状动脉灌注压、冠状动脉渗漏和组织水肿形成。胶体的额外应用增强了抗凝血酶的这些作用。电子显微镜显示抗凝血酶治疗后糖萼基本完整，抗凝血酶保护了内皮糖萼，维持了血管屏障功能，减少了间质水肿。胶体在这些心脏中的增强作用表明，防止脱落在体内也应该具有功能益处。
Antithrombin is an important inhibitor of the coagulation system, additionally exerting specific anti-inflammatory effects on endothelial cells. Healthy vascular endothelium is coated by the endothelial glycocalyx, diminution of which increases capillary permeability, e.g. after ischaemia. Antithrombin is known to infiltrate the glycocalyx, binding to glycosaminoglycans, and to preserve the glycocalyx after application tumour necrosis factor-alpha. We investigated the influence of antithrombin on glycocalyx subjected to ischaemia/reperfusion.Isolated guinea pig hearts were perfused with Krebs-Henseleit buffer (KHB). Antithrombin was applied to achieve physiological levels (1 U/mL) before inducing 20 min of ischaemia (37 degrees C). Hearts were reperfused for 20 min at constant flow (baseline perfusion pressure 70 cmH(2)O) with KHB or KHB plus 2 g% hydroxyethyl starch (130 kDa). Coronary net fluid filtration was assessed directly by measuring transudate formation on the epicardial surface. Post-ischaemic coronary release of syndecan-1 and heparan sulfate was quantified by ELISA. Hearts were perfusion-fixed to visualize the glycocalyx by electron microscopy. Ischaemia/reperfusion caused degradation of the glycocalyx, enhanced coronary perfusion pressure, and increased vascular permeability. Antithrombin significantly reduced post-ischaemic glycocalyx shedding, coronary perfusion pressure, coronary leak, and tissue oedema formation compared to untreated hearts. Additional application of colloid augmented these actions of antithrombin. Electron microscopy revealed a mostly intact glycocalyx after antithrombin treatment.Antithrombin preserves the endothelial glycocalyx, sustaining the vascular barrier function and reducing interstitial oedema. The potentiated effect of colloid in these hearts suggests that the prevention of shedding should be of functional benefit also in vivo.