乙醇与急性心肌梗死后大鼠冠脉侧枝循环的关系及干预研究

发布时间：2018-01-02 15:40

  本文关键词：乙醇与急性心肌梗死后大鼠冠脉侧枝循环的关系及干预研究　出处：《山东大学》2014年硕士论文　论文类型：学位论文

  更多相关文章： 乙醇 心肌梗死 微血管密度 瑞舒伐他汀 VEGF 内皮抑素

【摘要】：背景及目的 心血管疾病是当前世界重要的死亡原因之一,家族史、年龄、高血压、高胆固醇血症、糖尿病、肥胖、饮酒等都是冠心病的危险因素。饮酒作为冠心病可以改变的危险因素之一,众多来自于大型队列研究及荟萃分析的流行病学研究表明,适量饮酒可以降低冠心病的风险,与其发病率、死亡率的关系呈“U”或“J”字形曲线,而大量饮酒会导致冠状动脉的钙化程度明显升高,使冠心病的发病率、死亡率明显增加。另外来自许多体外的实验研究显示少量饮酒可以明显提高内皮血管内皮生长因子(VEGF)的表达,从而促进血管的新生,临床研究也发现长期少量饮酒的冠心病的患者具有丰富的冠脉侧枝循环。然而长期给予不同乙醇量对急性心肌梗死后缺血心肌内血管新生的影响及其作用机制如何?目前尚缺乏这方面的研究。 羟甲基戊二酰辅酶A还原酶抑制剂(他汀)目前已是冠心病一级及二级预防的基石,临床证据已经充分表明,长期给予他汀类药物治疗能够显著降低心血管事件发生的风险,长期应用他汀的患者将明显获益,目前认为其机制主要通过降脂、抗炎、抗氧化、保护血管内皮细胞等作用发挥心血管保护功能。最新的研究还发现,他汀类药物可以抑制心室重构,并能够促进缺血心肌内血管新生等心脏保护作用。因此我们提出假设,若长期大量饮酒对心肌梗死后缺血心肌内冠脉侧枝循环有抑制作用,那么长期预防性应用他汀类药物是否能够逆转大量饮酒对血管新生的这种抑制作用?目前缺乏这方面的报道。 因此本实验首先利用急性心肌梗死动物模型观察长期不同量乙醇摄入对缺血心肌内血管新生及其血管生长调节因子(包括VEGF和内皮抑素)表达水平的影响,然而根据实验结果来验证提出的假设是否成立,若假设成立,则应用他汀类药物看是否可以逆转大量饮酒对心肌血管新生的抑制作用,旨在为新的冠心病干预方式提供更多的理论支持。 第一部分乙醇对急性心肌梗死后大鼠冠脉侧枝循环的影响 方法 选取225-275g雄性SD大鼠,适应性喂养一周后；随机分成4组：假手术组(不做任何干预处理),空白对照组(蒸馏水,5g·kg·d-1),乙醇干预组：小剂量(50%食用乙醇,0.5g·kg-1·d-1),大剂量(50%食用乙醇,5g·kg-1·d-1)。连续喂养4周后制作成心肌梗死模型,假手术组在相应部位空穿一次；于术后第3天处死所有组别内的大鼠,留取梗死边缘区心肌组织,采用免疫组织化学SP法检测梗死边缘区心肌内CD34微血管密度(MVD),应用ELISA法测定梗死边缘区心肌组织匀浆液中血管内皮生长因子(VEGF)及内皮抑素的表达水平。通过方差分析研究各组间梗死边缘区MVD及VEGF.内皮抑素表达水平有无差异。 结果显示 空白对照组与假手术组相比,MVD明显增加[(88.7±3.4)N/mm2比(70.6±6.3)N/mm2,P0.05)]；VEGF.内皮抑素水平也明显升高[(318.7±18.6)pg/ml比(217.4±26.5)pg/ml；(306.9±53.9)ng/ml比(194.1±11.3)ng/ml,P0.05)]；与空白对照组相比,小剂量乙醇组MVD明显增加[(128.6±12.9)N/mm2比(88.7±3.4)N/mm2,P0.05)],VEGF表达明显升高[(438.2±46.2)pg/ml比(318.7±18.6)pg/ml,P0.05)],而内皮抑素浓度明显降低[(230.7±48.1)ng/ml比(306.9±53.9)ng/ml,P0.05)],而大剂量乙醇组与空白对照组相比,MVD明显减少[(55.4±8.6)N/mm2比(88.7±3.4)N/mm2,P0.05)],VEGF水平虽无明显差异,但内皮抑素表达水平明显升高[(437.5±66.7)ng/ml比(306.9±53.9)ng/ml,P0.05)]。 第二部分瑞舒伐他汀改善大量乙醇对急性心肌梗死后大鼠血管新生的影响 方法 选取225-275g雄性SD大鼠,设立瑞舒伐他汀预处理组：瑞舒伐他汀[(20mg·kg-1·d-1)+大剂量乙醇组(50%食用乙醇,5g·kg-1·d-1)],与第一部分的空白对照组(蒸馏水5g·kg-1·d-1)及大剂量乙醇组(50%乙醇,5g·kg-1·d-1)相比较,所有组别内大鼠连续喂养4周后,制作心肌梗死动物模型,术后第3天处死所有组别大鼠,留取梗死边缘区存活心肌组织,采用免疫组织化学SP法检测梗死边缘区心肌内CD34微血管密度(MVD),应用ELISA法测定各组梗死边缘区心肌组织匀浆液中血管内皮生长因子(VEGF)及内皮抑素水平。 结果显示 大剂量乙醇组与空白对照组相比的结果同第一部分,与大剂量乙醇组相比,瑞舒伐他汀预处理组MVD明显增加[(73.1±3.2)N/mm2比(55.4±8.6)N/mm2,P0.05)]; VEGF表达略有升高,内皮抑素有降低趋势,但与大量乙醇组相比均无明显统计学差异。 结论 1.长期饮酒可以影响急性心肌梗死后缺血心肌内VEGF、内皮抑素的表达 2.少量饮酒通过促进VEGF,并抑制内皮抑素的表达促进冠脉侧枝循环血管的新生； 3.长期大量饮酒可抑制急性心肌梗死后缺血心肌的血管新生,作用机制可能与增加内皮抑素的表达有关； 4.预防性给予瑞舒伐他汀能够改善长期大量饮酒后对心肌血管新生的抑制作用,但与VEGF和内皮抑素表达无关；
[Abstract]:Background and purpose
Cardiovascular disease is a major cause of death in the world, family history, age, hypertension, hypercholesterolemia, diabetes, obesity, alcohol consumption is a risk factor for coronary heart disease. One of the risk factors of coronary heart disease as drinking can change, many from large cohort studies and meta-analysis of epidemiological studies showed that moderate drinking can reduce coronary heart disease the risk of mortality and morbidity, the relationship of a "U" or "J" shaped curve, a large number of drinking will lead to the calcification of coronary artery was increased, the incidence of coronary heart disease, mortality increased significantly. Experimental studies also showed in vitro from many small amounts of alcohol can significantly improve endothelial vascular endothelial growth factor (VEGF) expression, so as to promote angiogenesis, clinical studies also showed that the long-term drinking of the coronary heart disease patients with coronary collateral rich However, the effect of long-term ethanol dosage on angiogenesis in ischemic myocardium after acute myocardial infarction is still unclear.
HMG CoA reductase inhibitor A two (Ta Ting) is now a grade and two grade prevention of coronary heart disease and clinical evidence has indicated that chronic administration of Ta Ting drug treatment can significantly reduce the risk of cardiovascular events, long-term application of Ta Ting's patients will benefit significantly, it is suggested that the main mechanism by lipid-lowering, anti-inflammatory antioxidation and protection of vascular endothelial cells, play the role of the function of cardiovascular protection. Recent studies also found that Ta Ting drugs can inhibit ventricular remodeling, and can promote the myocardial protective effect of angiogenesis in the heart. So we propose the hypothesis that if a large number of long-term drinking of myocardial infarction after myocardial ischemia in coronary collateral circulation inhibition so, whether the application of long-term prevention of Ta Ting's drugs to reverse a large number of drinking this inhibitory effect on the angiogenesis of the lack of this? Report.
Therefore this experiment using an animal model of acute myocardial infarction on different long-term ethanol intake regulation factor on angiogenesis in ischemic myocardium and vascular growth (including VEGF and endostatin) expression levels, however, according to the experimental results to verify the proposed hypotheses, if the hypothesis is to see whether the use of statins can be reversed drinking a lot of inhibitory effects on myocardial angiogenesis, in order to provide more theoretical support for the new way of coronary intervention.
The effect of part one ethanol on coronary collateral circulation in rats after acute myocardial infarction
Method
Male 225-275g SD rats, feeding a week; randomly divided into 4 groups: sham operation group (without any treatment), control group (distilled water, 5g - kg - D-1), ethanol intervention group: low dose (50% ethanol, 0.5g - kg-1 - D-1), high dose (50% ethanol, 5g - kg-1 - D-1). After 4 weeks of continuous feeding into the model of myocardial infarction in a sham operation group in the corresponding position; on the third day after surgery were all in rats, the myocardial tissue edge area of infarction, using immunohistochemical SP method to detect myocardial infarction edge CD34 in the area of microvessel density (MVD), vascular endothelial growth factor in myocardial infarction tissue homogenate in marginal zone by ELISA (VEGF) and the expression of endostatin. Through the analysis of variance the difference between the expression of MVD and VEGF. in infarcted zone between each level of endostatin.
Results show
绌虹櫧瀵圭収缁勪笌鍋囨墜鏈�缁勭浉姣�,MVD鏄庢樉澧炲姞[(88.7卤3.4)N/mm2姣，

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