Supplementary Material for: High Levels of Soluble Endoglin Induce a Proinflammatory and Oxidative-Stress Phenotype Associated with Preserved NO-Dependent Vasodilatation in Aortas from Mice Fed a High-Fat Diet

<b><i>Aims:</i></b> A soluble form of endoglin (sEng) was proposed to participate in the induction of endothelial dysfunction in small blood vessels. Here, we tested the hypothesis that high levels of sEng combined with a high-fat diet induce endothelial dysfunction in an atherosclerosis-prone aorta. <b><i>Methods and Results:</i></b> Six-month-old female and male transgenic mice overexpressing human sEng (<i>Sol-Eng</i>^<i>+</i>) with low (<i>Sol-Eng</i>^<i>+</i><i>low</i>) or high (<i>Sol-Eng</i>^<i>+</i><i>high</i>) levels of plasma sEng were fed a high-fat rodent diet containing 1.25% cholesterol and 40% fat for 3 months. The plasma cholesterol and mouse sEng levels did not differ in the <i>Sol-Eng</i>^<i>+</i><i>high</i> and <i>Sol-Eng</i>^<i>+</i><i>low</i> mice. The expression of proinflammatory (P-selectin, ICAM-1, pNFκB and COX-2) and oxidative-stress-related markers (HO-1, NOX-1 and NOX-2) in the aortas of <i>Sol-Eng</i>^<i>+</i><i>high</i> female mice was significantly higher than in <i>Sol-Eng</i>^<i>+</i><i>low </i>female mice. Endothelium-dependent vasodilatation induced by acetylcholine was preserved better in the <i>Sol-Eng</i>^{<i>+ </i>}<i>high</i> female mice than in the <i>Sol-Eng</i>^<i>+</i><i>low </i>female mice. <b><i>Conclusion:</i></b> These results suggest that high concentrations of sEng in plasma in combination with a high-fat diet induce the simultaneous activation of proinflammatory, pro-oxidative and vasoprotective mechanisms in mice aorta and the balance of these biological processes determines whether the final endothelial phenotype is adaptive or maladaptive.

*研究目的*：有研究提出可溶性内皮糖蛋白（soluble endoglin, sEng）可参与诱导小血管内皮功能障碍。本研究旨在验证下述假说：高水平sEng联合高脂饮食，可在易患动脉粥样硬化的主动脉中诱发内皮功能障碍。
*方法与结果*：选取6月龄、过表达人源sEng的转基因雌雄小鼠，其血浆sEng水平分为低表达组（*Sol-Eng*⁺*low*）与高表达组（*Sol-Eng*⁺*high*），给予含1.25%胆固醇、40%脂肪的高脂啮齿类饲料喂养3个月。结果显示，*Sol-Eng*⁺*high* 与 *Sol-Eng*⁺*low* 小鼠的血浆胆固醇与小鼠内源sEng水平无显著差异。与*Sol-Eng*⁺*low* 雌性小鼠相比，*Sol-Eng*⁺*high* 雌性小鼠的主动脉组织中，促炎标志物（P-选择素、细胞间黏附分子1、磷酸化NFκB及环氧合酶2）与氧化应激相关标志物（血红素氧合酶1、NADPH氧化酶1及NADPH氧化酶2）的表达水平显著升高。乙酰胆碱诱导的内皮依赖性血管舒张功能在*Sol-Eng*⁺*high* 雌性小鼠中的保留程度优于*Sol-Eng*⁺*low* 雌性小鼠。
*研究结论*：上述结果表明，血浆中高浓度sEng联合高脂饮食，可同时激活小鼠主动脉内的促炎、促氧化与血管保护生物学通路；这些过程的动态平衡，决定了最终内皮表型为适应性改变还是病理性失调。