Gordon Scott M, McKenzie Benjamin, Kemeh Georgina, Sampson Maureen, Perl Shira, Young Neal S, Fessler Michael B, Remaley Alan T
From the ‡Lipoprotein Metabolism Section, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland;
§Cell Biology Section, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland;
Mol Cell Proteomics. 2015 Dec;14(12):3247-57. doi: 10.1074/mcp.M115.054031. Epub 2015 Oct 19.
Statins lower plasma cholesterol by as much as 50%, thus reducing future cardiovascular events. However, the physiological effects of statins are diverse and not all are related to low density lipoprotein cholesterol (LDL-C) lowering. We performed a small clinical pilot study to assess the impact of statins on lipoprotein-associated proteins in healthy individuals (n = 10) with normal LDL-C (<130 mg/dL), who were treated with rosuvastatin (20 mg/day) for 28 days. Proteomic analysis of size-exclusion chromatography isolated LDL, large high density lipoprotein (HDL-L), and small HDL (HDL-S) fractions and spectral counting was used to compare relative protein detection before and after statin therapy. Significant protein changes were found in each lipoprotein pool and included both increases and decreases in several proteins involved in lipoprotein metabolism, complement regulation and acute phase response. The most dramatic effect of the rosuvastatin treatment was an increase in α-1-antirypsin (A1AT) spectral counts associated with HDL-L particles. Quantitative measurement by ELISA confirmed an average 5.7-fold increase in HDL-L associated A1AT. Molecular modeling predictions indicated that the hydrophobic reactive center loop of A1AT, the functional domain responsible for its protease inhibitor activity, is likely involved in lipid binding and association with HDL was found to protect A1AT against oxidative inactivation. Cell culture experiments, using J774 macrophages, demonstrated that the association of A1AT with HDL enhances its antiprotease activity, preventing elastase induced production of tumor necrosis factor α. In conclusion, we show that statins can significantly alter the protein composition of both LDL and HDL and our studies reveal a novel functional relationship between A1AT and HDL. The up-regulation of A1AT on HDL enhances its anti-inflammatory functionality, which may contribute to the non-lipid lowering beneficial effects of statins.
他汀类药物可使血浆胆固醇降低多达50%，从而减少未来心血管事件的发生。然而，他汀类药物的生理作用是多样的，并非所有作用都与降低低密度脂蛋白胆固醇（LDL-C）有关。我们进行了一项小型临床试点研究，以评估他汀类药物对LDL-C正常（<130 mg/dL）的健康个体（n = 10）脂蛋白相关蛋白的影响，这些个体接受瑞舒伐他汀（20 mg/天）治疗28天。采用排阻色谱法分离LDL、大高密度脂蛋白（HDL-L）和小HDL（HDL-S）组分，并通过蛋白质组学分析和光谱计数比较他汀类药物治疗前后的相对蛋白质检测情况。在每个脂蛋白池中均发现了显著的蛋白质变化，包括参与脂蛋白代谢、补体调节和急性期反应的几种蛋白质的增加和减少。瑞舒伐他汀治疗最显著的效果是与HDL-L颗粒相关的α-1-抗胰蛋白酶（A1AT）光谱计数增加。ELISA定量测量证实，与HDL-L相关的A1AT平均增加了5.7倍。分子模型预测表明，A1AT的疏水反应中心环（负责其蛋白酶抑制剂活性的功能域）可能参与脂质结合，并且发现与HDL的结合可保护A1AT免受氧化失活。使用J774巨噬细胞进行的细胞培养实验表明，A1AT与HDL的结合增强了其抗蛋白酶活性，可防止弹性蛋白酶诱导肿瘤坏死因子α的产生。总之，我们表明他汀类药物可显著改变LDL和HDL的蛋白质组成，我们的研究揭示了A1AT与HDL之间一种新的功能关系。HDL上A1AT的上调增强了其抗炎功能，这可能有助于他汀类药物的非降脂有益作用。
