α-亚麻酸植物甾醇酯对动脉粥样硬化的保护作用及其机制研究

发布时间：2018-06-12 03:22

  本文选题：动脉粥样硬化 + α-亚麻酸植物甾醇酯　； 参考：《华中科技大学》2015年博士论文

【摘要】：目的：探讨α-亚麻酸植物甾醇酯(a-linolenic acid ester of plant sterol, ALA-PS)对高脂饲料喂养载脂蛋白E基因敲除(apol ipoprote i n E knockout, ApoE KO)小鼠动脉粥样硬化(atherosclerosis, AS)的保护作用及相关分子机制。 方法：将42只ApoE KO小鼠(6周龄,雄性,体重12-15g)随机分为3组,即高脂饲料组(high fat diet, HFD),亚麻油干预组(flaxseed oil, FO)和亚麻油添加a-亚麻酸植物甾醇酯干预组(FO+ALA-PS)。 HFD组给予高脂饲料(脂肪21%,胆固醇0.15%,蛋白质200%)：FO组给予相同的高脂饲料和5%亚麻油；FO+ALA-PS组给予相同的高脂饲料并添加2.6%亚麻油和3.3%ALA-PS。对照组(Control)为14只相同遗传背景下的野生型C57BL/6小鼠,给予普通饲料喂养,实验周期为18周。 结果： 1、主动脉整体、主动脉窦和主动脉弓油红O及HE染色结果显示,与对照组小鼠相比,HFD组小鼠血管内壁可见大面积AS斑块形成。ALA-PS干预后显著减少AS斑块面积(P0.05)。免疫组织化学染色结果显示,ALA-PS明显改善HFD诱导的巨噬细胞和单核细胞聚集浸润以及血管平滑肌细胞增殖与迁移。 2、HFD诱导脂代谢紊乱。ALA-PS干预显著改善血脂异常以及肝脏总胆固醇(total cholesterol, TC)和甘油三酯(triglyceride, TG)水平升高(P0.05)。并且,明显升高肝脏低密度脂蛋白受体(LDLr)、清道夫受体(SR-BI)和胆固醇外流关键因子腺苷三磷酸结合核转运受体Al (ABCA1)、肝脏X受体-α(LXRa)mRNA和蛋白的表达水平以及明显降低小鼠肝脏胆固醇合成关键因子羟甲基戊二酸单酰辅酶A还原酶(HMGCR)和胆固醇调节元件结合蛋白-2(SREBP-2) mRNA和蛋白的表达水平(P0.05)。此外,ALA-PS干预显著增加脂肪酸β-氧化关键因子过氧化物酶体增殖物激活受体-α (PPARa)、肉毒碱棕榈酰基转移酶1A (CPT1A)和乙酰辅酶A氧化酶1(ACOXI) mRNA和蛋白的表达水平,并显著降低调控脂质新生关键因子胆固醇调节元件结合蛋白-1(SREBP-1)和乙酰辅酶A羧化酶(ACC) mRNA和蛋白的表达水平(P0.05)。 3、HFD诱导炎症反应增加。ALA-PS干预能够显著降低血浆炎症因子IL-1β、IL-6. TNF-α、 MCP-1、 sVCAM-1和sIC AM-1的水平,并且明显抑制小鼠主动脉IL-1β、 IL-6、 TNF-α、 MCP-1、 VCAM-1和ICAM-1mRNA和蛋白的表达水平升高(P0.05)。同时,显著降低小鼠外周血单核细胞IL-1β、 IL-6、 TNF-a和MCP-1mRNA和蛋白的表达水平(P0.05)。 4、HFD诱导机体氧化应激。ALA-PS干预能够有效地降低血清和肝脏MDA含量并增加GSH水平,同时显著抑制主动脉ROS产生(P0.05)。并且,ALA-PS干预显著减少小鼠主动脉NADPH氧化酶亚基P22phox、 p47phox、 p67phox和gp91phox mRNA和蛋白表达(P0.05)。 结论：α-亚麻酸植物甾醇酯对动脉粥样硬化具有保护作用,其机制与改善脂代谢、调节炎症反应以及抑制氧化应激有关。
[Abstract]:Objective: to investigate the protective effect of 伪 -linolenic acid ester of plant sterol, ALA-PS) on Apolipoprotein E knockout (ApoE KOA) gene knockout (ApoEKOA) mice fed high fat diet. Methods: the protective effect of 伪 -linolenic acid ester of plant sterol, ALA-PSN on atherosclerotic atherosclerosis (ASA) in mice fed with high fat diet was studied. : 42 ApoE KO mice aged 6 weeks, Male, weight 12-15 g) were randomly divided into three groups: high fat dietate, HFDD, flax seed oil (FOO) and FO ALA-PSN treated with flax oil. The HFD group was given the same high fat diet (fat 21, cholesterol 0.15, protein 200: FO) and 5% linseed oil FO ALA-PS with the same high fat diet and 2.6% linseed oil and 3.3 ALA-PSs. The control group was 14 wild-type C57BL / 6 mice with the same genetic background, fed with common diet for 18 weeks. Results: 1. The results showed that: 1, the whole aorta, aortic sinus and aortic arch were stained with oil red O and HE. Compared with the control group, large area of as plaque formation was observed in the intravascular wall of HFD group. ALA-PS significantly decreased as plaque area (P0.05) after intervention. The results of immunohistochemical staining showed that ALA-PS significantly improved the aggregation and infiltration of macrophages and monocytes induced by HFD and the proliferation and migration of vascular smooth muscle cells. Total cholesterol (TCc) and triglyceride (TGG) levels increased P0.05. And, The expression of LDLrN, SR-BI), the key factor of cholesterol efflux, adenosine triphosphate binding nuclear transport receptor, the expression of liver X receptor-伪 -LXRaN mRNA and protein, and the decrease of liver bile duct in mice were significantly increased, and the key factors of cholesterol efflux, adenosine triphosphate binding nuclear transport receptor, liver X receptor-伪 -LXRaN mRNA and protein expression were significantly increased. The expression levels of HMGCR-HMGCRand cholesterol regulatory element binding protein-SREBP-2 mRNA and protein in steroid synthesis key factor hydroxymethyl glutaric acid monoacyl coA reductase A reductase (HMGCR-HMGCRA) and cholesterol regulatory element binding protein-SREBP-2 were found to be P0.05a. In addition, ALA-PS significantly increased the expression levels of PPARaA, carnitine palmitoyltransferase 1A (CPT1A) and acetyl-coA oxidase 1 (ACOXI) mRNA and protein in peroxisome proliferator activator receptor (PPARaA) and acetyl coA oxidase (ACOA oxidase 1). The levels of mRNA and protein expression of cholesterol regulatory element binding protein-1 (SREBP-1) and acetyl-coA carboxylase (ACC-1) were significantly decreased. The levels of TNF- 伪, MCP-1, sVCAM-1 and sIC AM-1 were significantly inhibited, and the expression levels of IL-1 尾, IL-6, TNF- 伪, MCP-1, VCAM-1 and ICAM-1 mRNA and protein in aorta of mice were significantly increased (P 0.05). At the same time, the expression of IL-1 尾, IL-6, TNF-a and MCP-1 mRNA and protein in peripheral blood monocytes of mice was significantly decreased. 4. The intervention of HFD induced by oxidative stress could effectively reduce the MDA content and increase the level of GSH in serum and liver, and significantly inhibit the production of P0.05 by aortic Ros. ALA-PS significantly reduced the expression of NADPH oxidase subunits P22phox, p47phox, p67phox and gp91phox mRNA and protein in mouse aorta. Conclusion: phytosterol 伪 -linolenic acid has protective effect on atherosclerosis, and its mechanism and lipid metabolism are improved. Regulation of inflammation and inhibition of oxidative stress.
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R151
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本文编号：2008074