Procedures to Evaluate the Role of Heparan Sulfate on the Reactivity of Resistance and Conductance Arteries Ex Vivo.
Procedures to Evaluate the Role of Heparan Sulfate on the Reactivity of Resistance and Conductance Arteries Ex Vivo.
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DOI：
10.1007/978-1-0716-1398-6_40
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发表时间：
2022
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越来越多的证据表明，内皮糖萼的破坏可能是糖尿病患者动脉功能障碍的重要原因。在不受神经、体液和机械影响的情况下，评估内皮和血管平滑肌细胞层完整性的一种方法是测量动脉血管的离体运动。在这里，我们描述了一个程序来评估非受体介导的血管收缩，受体介导的血管收缩，内皮依赖性和非依赖性血管扩张，在阻力和传导动脉加压至60毫米汞柱。除了用等压法评估血管反应性外，同样的实验装置可以用来启动跨动脉的压力梯度，这样就可以测量腔内血流介导的血管舒张。在使用等压或流动介导的方法记录内皮依赖性血管舒张后，可以在酶的存在下完成相同的干预，这些酶将具有生物活性的硫酸肝素切割成无活性的双糖和低聚糖片段，以评估来自：(a)内皮衍生物质（例如，通过一氧化氮合酶抑制的一氧化氮）的贡献；或(b)糖萼的重要成分（例如，通过肝素酶III处理去除硫酸肝素）。在这里，我们表明，急性破坏主要的糖胺聚糖，即硫酸肝素，损害小鼠阻力动脉腔内血流介导的血管舒张。
Evidence is emerging that disruption of the endothelial glycocalyx might contribute importantly to arterial dysfunction in the context of diabetes. One approach to assess the integrity of the endothelium and the vascular smooth muscle cell layer, in the absence of neural, humoral, and mechanical influences, is by measuring arterial vasomotion ex vivo. Here we describe a procedure to assess non-receptor-mediated vasoconstriction, receptor-mediated vasoconstriction, and endothelium-dependent and -independent vasodilation, in resistance and conductance arteries pressurized to 60 mmHg. In addition to evaluating vasoreactivity using isobaric approaches, the same experimental set-up can be used to initiate a pressure gradient across the artery such that intraluminal, flow-mediated vasodilation can be measured. After recording endothelium-dependent vasodilation using isobaric or flow-mediated approaches, identical interventions can be completed in the presence of enzymes that cleave biologically active heparan sulfates into inactive disaccharide and oligosaccharide fragments to assess the contribution from: (a) endothelial-derived substances (e.g., nitric oxide via nitric oxide synthase inhibition); or (b) important components of the glycocalyx (e.g., removal of heparan sulfate via heparitinase III treatment). Here, we show that acute disruption of a predominant glycosaminoglycan i.e., heparan sulfate impairs intraluminal flow-mediated vasodilation in murine resistance arteries.