Lambert Carmen, Cubedo Judit, Padró Teresa, Sánchez-Hernández Joan, Antonijoan Rosa M, Perez Antonio, Badimon Lina
Cardiovascular Science Institute-ICCC IIB-Sant Pau, 08025 Barcelona, Spain.
Ciber CV, 28029 Madrid, Spain.
Nutrients. 2017 Jun 13;9(6):599. doi: 10.3390/nu9060599.
Cardiovascular disease (CVD) remains one of the major causes of death and disability worldwide. In addition to drug treatment, nutritional interventions or supplementations are becoming a health strategy for CVD prevention. Phytosterols (PhyS) are natural components that have been shown to reduce cholesterol levels; while poly-unsaturated fatty acids (PUFA), mainly omega-3 (ω3) fatty acids, have shown to reduce triglyceride levels. Here we aimed to investigate whether the proteins in the main lipoproteins (low density lipoproteins (LDL) and high density lipoproteins (HDL)) as well as proteins in the lipid free plasma fraction (LPDP) were regulated by the intake of PhyS-milk or ω3-milk, in overweight healthy volunteers by a proteomic based systems biology approach. The study was a longitudinal crossover trial, including thirty-two healthy volunteers with body mass index (BMI) 25-35 kg/m² (Clinical Trial: ISRCTN78753338). Basal samples before any intervention and after 4 weeks of intake of PhyS or ω3-milk were analyzed. Proteomic profiling by two dimensional electrophoresis (2-DE) followed by mass spectrometry-(MALDI/TOF), ELISA, Western blot, conventional biochemical analysis, and in-silico bioinformatics were performed. The intake of PhyS-milk did not induce changes in the lipid associated plasma protein fraction, whereas ω3-milk significantly increased apolipoprotein (Apo)- E LDL content ( 0.043) and induced a coordinated increase in several HDL-associated proteins, Apo A-I, lecitin cholesterol acyltransferase (LCAT), paraoxonase-1 (PON-1), Apo D, and Apo L1 ( < 0.05 for all). Interestingly, PhyS-milk intake induced a reduction in inflammatory molecules not seen after ω3-milk intake. Serum amyloid P component (SAP) was reduced in the LPDP protein fraction ( = 0.001) of subjects taking PhyS-milk and C-C motif chemokine 2 (CCL2)expression detected by reverse transcription polymerase chain reaction (RT-PCR) analysis in white blood cells was significantly reduced ( = 0.013). No changes were observed in the lipid-free plasma proteome with ω3-milk. Our study provides novel results and highlights that the PhyS-milk induces attenuation of the pro-inflammatory pathways, whereas ω3-milk induces improvement in lipid metabolic pathways.
心血管疾病（CVD）仍然是全球主要的死亡和残疾原因之一。除药物治疗外，营养干预或补充正成为预防CVD的一项健康策略。植物甾醇（PhyS）是已被证明可降低胆固醇水平的天然成分；而多不饱和脂肪酸（PUFA），主要是ω-3（ω3）脂肪酸，已被证明可降低甘油三酯水平。在此，我们旨在通过基于蛋白质组学的系统生物学方法，研究超重健康志愿者摄入含植物甾醇牛奶或ω3牛奶后，主要脂蛋白（低密度脂蛋白（LDL）和高密度脂蛋白（HDL））中的蛋白质以及无脂血浆部分（LPDP）中的蛋白质是否受到调控。该研究是一项纵向交叉试验，纳入了32名体重指数（BMI）为25 - 35 kg/m²的健康志愿者（临床试验：ISRCTN78753338）。分析了任何干预前的基础样本以及摄入植物甾醇或ω3牛奶4周后的样本。通过二维电泳（2-DE）进行蛋白质组分析，随后进行质谱分析（MALDI/TOF）、酶联免疫吸附测定（ELISA）、蛋白质印迹法、传统生化分析以及计算机生物信息学分析。摄入含植物甾醇牛奶未引起与脂质相关的血浆蛋白部分发生变化，而ω3牛奶显著增加了载脂蛋白（Apo）-E LDL含量（P = 0.043），并导致几种与HDL相关的蛋白质协同增加，包括Apo A-I、卵磷脂胆固醇酰基转移酶（LCAT）、对氧磷酶-1（PON-1）、Apo D和Apo L1（所有P均<0.05）。有趣的是，摄入含植物甾醇牛奶导致炎症分子减少，而摄入ω3牛奶后未出现这种情况。摄入含植物甾醇牛奶的受试者的LPDP蛋白部分中的血清淀粉样P成分（SAP）减少（P = 0.001），通过逆转录聚合酶链反应（RT-PCR）分析检测到白细胞中C-C基序趋化因子2（CCL2）的表达显著降低（P = 0.013）。摄入ω3牛奶后无脂血浆蛋白质组未观察到变化。我们的研究提供了新的结果，并突出表明含植物甾醇牛奶可诱导促炎途径减弱，而ω3牛奶可诱导脂质代谢途径改善。
