黄酒中抗低密度脂蛋白受体敲除小鼠动脉粥样硬化成分研究

STUDY ON THE ANTI-ATHEROSCLEROTIC COMPONENTS IN CHINESE YELLOW WINE IN LOW DENSITY LIPOPROTEIN RECEPTOR KNOCKOUT MICE

目的探索黄酒中具有抑制高脂饮食诱导低密度脂蛋白受体敲除(LDLR-/-)小鼠动脉粥样硬化作用的活性成分。方法 48只6w龄雄性LDLR-/-小鼠,随机分为6组:高脂对照组、黄酒低聚糖干预组、黄酒多肽干预组、黄酒多酚干预组、酒精干预组、黄酒干预组,每组8只。干预16w后处死,分离血清,检测血脂含量,苏丹Ⅳ染色后观察胸腹主动脉粥样斑块大小,Western blot测定主动脉组织内基质金属蛋白酶2(matrix metalloproteinase2,MMP-2)、基质金属蛋白酶9(matrix metalloproteinase 9,MMP-9)以及组织型基质金属蛋白酶抑制剂2(tissue inhibitor of metalloproteinase2,TIMP-2)的表达,以明胶酶谱法测定主动脉弓动脉粥样硬化处MMP-2和MMP-9的活性。结果相比于对照组,黄酒多酚干预组、黄酒干预组小鼠低密度脂蛋白(low density lipoprotein cholesterol,LDL-C)、甘油三酯(triglyceride,TG)、总胆固醇(total cholesterol,TC)水平(P<0.05),黄酒多肽干预组小鼠LDL-C、TC水平降低(P<0.05)。相比于对照组,黄酒多酚、黄酒多肽、黄酒干预组小鼠主动脉斑块面积明显缩小(P<0.05)。相比于高脂组,黄酒多肽、黄酒多酚、黄酒干预组小鼠MMP-2/9表达减少、活性降低(P<0.05)。各组小鼠TIMP-2的表达没有明显影响。结论黄酒中的多肽类和多酚类成分具有抑制高脂饮食诱导的LDLR-/-小鼠动脉粥样硬化的作用,其机制可能是通过改善小鼠血脂情况,维持主动脉组织中MMPs与TIMPs之间的平衡。

Objective To explore the active ingredients in Chinese yellow wine which could inhibit the progress of atherosclerosis in low density lipoprotein receptor （LDLR） knockout mice. Methods Six weeks old LDLR-/- male mice （n=48） were randomly allocated into 6 groups （8 mice in each group）： high fat group （control group）, yellow wine oligosaccharides modulation group, yellow wine polypeptides modulation group, yellow wine polyphenols modulation group, alcohol modulation group, yellow wine modulation group. After 16 weeks intervention, mice were sacrificed. The levels of serum lipids were examined. The morphological changes of aorta atheromatous plaque were observed under microscope. The expressions of matrix metalloproteinase 2（MMP-2）, matrix metalloproteinase 9 （MMP-9） and tissue inhibitor of metalloproteinase2 （TIMP-2） were determined by Western blot. The activations of MMP-2, MMP-9 were determined by Gelatin Zymography. Results Compared with the control group, the level of LDL-C, TG and TC in the serum were decreased in yellow wine polyphenols group and yellow wine group （P〈0.05）; the level of LDL-C and TC in the serum were decreased in polypeptides group （P〈0.05）. Compared with the control group, the atherosclerosis lesion area were significantly smaller in polypeptides, polyphenols and yellow wine groups （P〈0.05）. Compared with the control group, the expression and activation of MMP-2 and MMP-9 were decreased in polypeptides, polyphenols and yellow wine groups （P〈0.05）. There was no significant difference among each group on the expression of TIMP-2. Conclusion Polypeptides and polyphenols in the Chinese yellow wine could inhibit the high fat diet induced progression of atherosclerosis in LDLR-/- mice. The mechanisms may be that these ingredients in the wine could regulate blood lipid metabolism and keep the balance between MMPs and TIMPs.