Biochemistry and Biotechnology Department, Universitat Rovira i Virgili, 43007 Tarragona, Spain.
Mol Cell Proteomics. 2010 Jul;9(7):1499-513. doi: 10.1074/mcp.M000055-MCP201. Epub 2010 Mar 23.
Bioactive proanthocyanidins have been reported to have several beneficial effects on health in relation to metabolic syndrome, type 2 diabetes, and cardiovascular disease. We studied the effect of grape seed proanthocyanidin extract (GSPE) in rats fed a high fat diet (HFD). This is the first study of the effects of flavonoids on the liver proteome of rats suffering from metabolic syndrome. Three groups of rats were fed over a period of 13 weeks either a chow diet (control), an HFD, or a high fat diet supplemented for the last 10 days with GSPE (HFD + GSPE). The liver proteome was fractionated, using a Triton X-114-based two-phase separation, into soluble and membrane protein fractions so that total proteome coverage was considerably improved. The data from isobaric tag for relative and absolute quantitation (iTRAQ)-based nano-LC-MS/MS analysis revealed 90 proteins with a significant (p < 0.05) minimal expression difference of 20% due to metabolic syndrome (HFD versus control) and 75 proteins due to GSPE treatment (HFD + GSPE versus HFD). The same animals have previously been studied (Quesada, H., del Bas, J. M., Pajuelo, D., Díaz, S., Fernandez-Larrea, J., Pinent, M., Arola, L., Salvadó, M. J., and Bladé, C. (2009) Grape seed proanthocyanidins correct dyslipidemia associated with a high-fat diet in rats and repress genes controlling lipogenesis and VLDL assembling in liver. Int. J. Obes. 33, 1007-1012), and GSPE was shown to correct dyslipidemia observed in HFD-fed rats probably through the repression of hepatic lipogenesis. Our data corroborate those findings with an extensive list of proteins describing the induction of hepatic glycogenesis, glycolysis, and fatty acid and triglyceride synthesis in HFD, whereas the opposite pattern was observed to a large extent in GSPE-treated animals. GSPE was shown to have a wider effect than previously thought, and putative targets of GSPE involved in the reversal of the symptoms of metabolic syndrome were revealed. Some of these novel candidate proteins such as GFPT1, CD36, PLAA (phospholipase A(2)-activating protein), METTL7B, SLC30A1, several G signaling proteins, and the sulfide-metabolizing ETHE1 and SQRDL (sulfide-quinone reductase-like) might be considered as drug targets for the treatment of metabolic syndrome.
生物活性原花青素已被报道具有多种有益的健康效应，与代谢综合征、2 型糖尿病和心血管疾病有关。我们研究了葡萄籽原花青素提取物（GSPE）对高脂肪饮食（HFD）喂养大鼠的影响。这是第一个研究黄酮类化合物对患有代谢综合征大鼠肝蛋白质组影响的研究。三组大鼠在 13 周内分别用标准饲料（对照）、高脂肪饮食或高脂肪饮食加最后 10 天补充 GSPE（HFD+GSPE）喂养。使用基于 Triton X-114 的两相分离法将肝蛋白质组分离为可溶性和膜蛋白部分，从而大大提高了总蛋白质组的覆盖率。基于等压标记相对和绝对定量（iTRAQ）的纳升 LC-MS/MS 分析的数据显示，由于代谢综合征（HFD 与对照），有 90 种蛋白质的最小表达差异具有显著意义（p < 0.05），差异为 20%，由于 GSPE 处理（HFD+GSPE 与 HFD），有 75 种蛋白质的最小表达差异具有显著意义。同一批动物以前曾被研究过（Quesada，H.，del Bas，J. M.，Pajuelo，D.，Díaz，S.，Fernández-Larrea，J.，Pinent，M.，Arola，L.，Salvadó，M. J.，和 Bladé，C.（2009）葡萄籽原花青素可纠正高脂肪饮食大鼠的血脂异常，并抑制控制肝内脂肪生成和 VLDL 组装的基因。国际肥胖杂志 33，1007-1012），并表明 GSPE 可纠正 HFD 喂养大鼠的血脂异常，可能是通过抑制肝内脂肪生成。我们的数据通过广泛的蛋白质组学描述，与这些发现相吻合，这些蛋白质描述了 HFD 诱导的肝糖生成、糖酵解、脂肪酸和甘油三酯合成，而在 GSPE 处理的动物中，观察到相反的模式。GSPE 的作用比以前认为的更广泛，并且揭示了与代谢综合征症状逆转有关的 GSPE 潜在靶点。其中一些新的候选蛋白质，如 GFPT1、CD36、PLAA（磷脂酶 A(2)-激活蛋白）、METTL7B、SLC30A1、几种 G 信号蛋白以及硫化物代谢 ETHE1 和 SQRDL（硫化物-醌还原酶样），可被视为代谢综合征治疗的药物靶点。
