Abstract：
OBJECTIVE: Inflammatory cells within atherosclerotic lesions secrete proteolytic enzymes that contribute to lesion progression and destabilization, increasing the risk for an acute cardiovascular event. Elastase is a serine protease, secreted by macrophages and neutrophils, that may contribute to the development of unstable plaque. We previously reported interaction of endogenous protease-inhibitor proteins with high-density lipoprotein (HDL), including alpha-1-antitrypsin, an inhibitor of elastase. These findings support a potential role for HDL as a modulator of protease activity. In this study, we test the hypothesis that enhancement of HDL-associated elastase inhibitor activity is protective against atherosclerotic lesion progression.
METHODS: We designed an HDL-targeting protease inhibitor (HTPI) that binds to HDL and confers elastase inhibitor activity. Lipoprotein binding and the impact of HTPI on atherosclerosis were examined using mouse models. Histology and immunofluorescence staining of aortic root sections were used to examine the impact of HTPI on lesion morphology and inflammatory features.
RESULTS: HTPI is a small (1.6 kDa) peptide with an elastase inhibitor domain, a soluble linker, and an HDL-targeting domain. When incubated with human plasma ex vivo, HTPI predominantly binds to HDL. Intravenous administration of HTPI to mice resulted in its binding to plasma HDL and increased elastase inhibitor activity on isolated HDL. Accumulation of HTPI within plaque was observed after administration to Apoe-/- mice. To examine the effect of HTPI treatment on atherosclerosis, prevention and progression studies were performed using Ldlr-/- mice fed Western diet. In both study designs, HTPI-treated mice had reduced lipid deposition in plaque.
CONCLUSIONS: These data support the hypothesis that HDL-associated anti-elastase activity can improve the atheroprotective potential of HDL and highlight the potential utility of HDL enrichment with anti-protease activity as an approach for stabilization of atherosclerotic lesions.
METHODS: We designed an HDL-targeting protease inhibitor (HTPI) that binds to HDL and confers elastase inhibitor activity. Lipoprotein binding and the impact of HTPI on atherosclerosis were examined using mouse models. Histology and immunofluorescence staining of aortic root sections were used to examine the impact of HTPI on lesion morphology and inflammatory features.
RESULTS: HTPI is a small (1.6 kDa) peptide with an elastase inhibitor domain, a soluble linker, and an HDL-targeting domain. When incubated with human plasma ex vivo, HTPI predominantly binds to HDL. Intravenous administration of HTPI to mice resulted in its binding to plasma HDL and increased elastase inhibitor activity on isolated HDL. Accumulation of HTPI within plaque was observed after administration to Apoe-/- mice. To examine the effect of HTPI treatment on atherosclerosis, prevention and progression studies were performed using Ldlr-/- mice fed Western diet. In both study designs, HTPI-treated mice had reduced lipid deposition in plaque.
CONCLUSIONS: These data support the hypothesis that HDL-associated anti-elastase activity can improve the atheroprotective potential of HDL and highlight the potential utility of HDL enrichment with anti-protease activity as an approach for stabilization of atherosclerotic lesions.
摘要：
目的：动脉粥样硬化病变内的炎症细胞分泌蛋白水解酶，有助于病变进展和不稳定，增加急性心血管事件的风险.弹性蛋白酶是一种丝氨酸蛋白酶,由巨噬细胞和嗜中性粒细胞分泌，这可能有助于不稳定斑块的发展。我们以前报道了内源性蛋白酶抑制剂蛋白与高密度脂蛋白（HDL）的相互作用，包括α-1-抗胰蛋白酶,弹性蛋白酶抑制剂.这些发现支持HDL作为蛋白酶活性调节剂的潜在作用。在这项研究中,我们检验了HDL相关弹性蛋白酶抑制剂活性增强对动脉粥样硬化病变进展具有保护作用的假设.
方法：我们设计了一种与HDL结合并赋予弹性蛋白酶抑制剂活性的HDL靶向蛋白酶抑制剂（HTPI）。使用小鼠模型检查脂蛋白结合和HTPI对动脉粥样硬化的影响。主动脉根部切片的组织学和免疫荧光染色用于检查HTPI对病变形态和炎症特征的影响。
结果：HTPI是一种具有弹性蛋白酶抑制剂结构域的小（1.6kDa）肽，可溶性接头，和HDL靶向域。当与人血浆离体孵育时，HTPI主要与HDL结合。向小鼠静脉内施用HTPI导致其与血浆HDL结合并增加了对分离的HDL的弹性蛋白酶抑制剂活性。在给予Apoe-/-小鼠后观察到HTPI在斑块内的积累。为了检查HTPI治疗对动脉粥样硬化的影响，使用喂食西方饮食的Ldlr-/-小鼠进行预防和进展研究。在两项研究设计中，HTPI处理的小鼠在斑块中具有减少的脂质沉积。
结论：这些数据支持以下假设：HDL相关的抗弹性蛋白酶活性可以改善HDL的动脉粥样硬化保护潜力，并强调了HDL富集与抗蛋白酶活性作为稳定动脉粥样硬化病变的方法的潜在效用。
方法：我们设计了一种与HDL结合并赋予弹性蛋白酶抑制剂活性的HDL靶向蛋白酶抑制剂（HTPI）。使用小鼠模型检查脂蛋白结合和HTPI对动脉粥样硬化的影响。主动脉根部切片的组织学和免疫荧光染色用于检查HTPI对病变形态和炎症特征的影响。
结果：HTPI是一种具有弹性蛋白酶抑制剂结构域的小（1.6kDa）肽，可溶性接头，和HDL靶向域。当与人血浆离体孵育时，HTPI主要与HDL结合。向小鼠静脉内施用HTPI导致其与血浆HDL结合并增加了对分离的HDL的弹性蛋白酶抑制剂活性。在给予Apoe-/-小鼠后观察到HTPI在斑块内的积累。为了检查HTPI治疗对动脉粥样硬化的影响，使用喂食西方饮食的Ldlr-/-小鼠进行预防和进展研究。在两项研究设计中，HTPI处理的小鼠在斑块中具有减少的脂质沉积。
结论：这些数据支持以下假设：HDL相关的抗弹性蛋白酶活性可以改善HDL的动脉粥样硬化保护潜力，并强调了HDL富集与抗蛋白酶活性作为稳定动脉粥样硬化病变的方法的潜在效用。
