普罗布考对高同型半胱氨酸血症致兔动脉粥样硬化的作用_图文

1、论著普罗布考对高同型半胱氨酸血症致兔动脉粥样硬化的作用吴乐a #,黎红华a ,武强a ,王晓昆b广州军区武汉总医院a.神经内科,b.医学实验中心,武汉430070【收稿日期】2010207207#【摘要】目的:观察普罗布考对高同型半胱氨酸(Hcy 血症致兔动脉粥样硬化(AS 的作用。方法:将24只大白兔分为对照组、模型组和治疗组各8只,对照组给予基础饲料喂养,模型组和治疗组均在基础饲料中加入1%的蛋氨酸,此外治疗组同时用普罗布考灌胃(1g/d ;饮食量200g/d ,均喂养20周。分别于第20周末检测血总同型半胱氨酸(t Hcy 、总胆固醇(TC 、甘油三酯(T G 、高密度脂蛋白胆固醇(HD

2、L 2C 和低密度脂蛋白胆固醇(LDL 2C 。第20周末处死动物,剥离腹主动脉,用Western blot 检测血管中核转录因子2B (N F 2B 蛋白表达,光镜下观察主动脉病理形态学变化。结果:治疗组血中TC 、T G 、HDL 2C 和LDL 2C 水平低于模型组(P 0.05,t Hcy水平无统计学差异;治疗组主动脉N F 2B 蛋白表达水平明显低于模型组(P 0.05;主动脉病理形态学检查显示治疗组病变轻于模型组。结论:普罗布考能降低血脂,下调N F 2B ,减轻内膜增生程度,具有抗AS 的作用。【关键词】普罗布考;同型半胱氨酸;动脉粥样硬化;核转录因子2BHong 2H ua ,

3、W U Qi ang ,W A N G X i ao 2k un.Dep artment of N eurolog y ,W uhan General Hos 2pit al of Guan gz hou M ilit ary Region ,Wuhan 430070,China【Abstract 】Objective :To observe t he effect of Probucol on at herosclerosis in rabbit s in 2duced by hyperhomocysteinemia.Met hods :Twenty 2four male rabbit s

4、were randomly divided into t hree group s.The cont rol group was fed on common food ;t he homocysterine (Hcy model group was fed on high 2met hionine food wit h 1%met hionine ;t he t reat ment group was fed wit h high 2me 2t hionine food and Probucol (1g/d perf used into stomach .Blood samples have

5、been collected on week 20after t he diet s and t he serum lipids and plasma total Hcy (t Hcy were measured.West 2ern 2blotting was used to investigate t he exp ression of nuclear factor 2B (N F 2B in aorta.The pat hological changes of t he aorta were examined under light microscopy.Result s :The lip

6、ids int he serum and t he expression of N F 2B were significantly reduced in t he t reat ment group relative to t ho se in t he Hcy model group.Aortic changes were also attenuated in treat ment group when compared to t hose in t he Hcy model group.Conclusion :Probucol could decrease serum lipids lev

7、 2el and reverse t he aortic at hero sclerotic changes induced by Hcy ,p robably by way of down 2regu 2lating N F 2B expression.【K ey w ords 】probucol ;homocysterine ;at herosclero sis ;nuclear factor 2B动脉粥样硬化(at hero sclero sis ,AS 是一种慢性炎症反应过程,主要累及中等以上的肌性动脉。核转录因子2B (nuclear fac 2tor 2B ,N F 2B 的激活是

8、AS 形成的重要环节。同型半胱氨酸(homocysterine ,Hcy 是导致AS的独立危险因素1,2。普罗布考(p robuco1已作为降脂药应用于临床,同时具有抗AS作用,推测可能与改善内皮功能有关3。本研究以高蛋氨酸饮食诱导兔高Hcy血症及AS模型,探讨普罗布考在高Hcy血症致AS过程中的作用。1材料与方法1.1材料动物:雄性纯种新西兰大白兔24只,体质量2.02.5kg,购自湖北省动物实验中心。主要试剂与材料:兔抗N F2B p65(购于美国Santa Cruz公司,辣根过氧化物酶标记的二抗(羊抗兔和羊抗鼠(购于武汉Bo ster公司,普罗布考粉剂(购于山东齐鲁制药有限公司。1.2方

9、法2结果治疗组较模型组血清血脂水平明显下降,但仍高于对照组,有统计学差异(P 0.05,模型组和治疗组血浆Hcy水平差异无统计学意义,见表1。Western blot显示,模型组N F2B p65蛋白表达较对照组显著增高(P=0.009。治疗组N F2B p65蛋白表达降低,与模型组相比差异有统计学意义(P=0.046,仍高于对照组(P=0.027,见图1A-B。H E染色显示对照组血管壁内膜光滑完整,中膜平滑肌细胞排列规则;模型组血管内膜明显增厚,可见一处内膜下出血(箭头处,中膜平滑肌细胞排列紊乱;治疗组血管内膜增厚明显减轻,中膜平滑肌细胞排列规则,见图2A-C。3讨论Schwartz等4研

10、究发现,普罗布考治疗后HDL颗粒的直径变小,数目不变,普罗布考治疗使HDL2C失去胆固醇部分而使单位体积血液中HDL2C的总重量降低。由于普罗布考在降胆固醇的同时降低HDL2C,而淡出市场。然而,临床研究发现,普罗布考虽然降低HDL2C,但显示出强大的抗AS、抑制再狭窄和改善血管舒张功能的作用5,6。表13组血脂及Hcy 水平比较(x -s 组别nTC/(mmol/L T G /(mmol/L HDL 2C/(mmol/L LDL 2C/(mmol/L Hcy/(86.631.781.190.211.090.185.271.1243.423.53与对照组比较,P 0.05,P 0.01;与模型

11、组比较,P 0.05 血浆Hcy 水平严重升高的患者常伴早发性AS 。血浆高Hcy 引起心血管疾病的机制未完全明了,目前认为主要与以下几个方面有关:导致内皮细胞损伤及功能障碍7,8;促进脂蛋白过氧化9;引起许多炎症介质释放10;增加肝细胞TC 合成等11;其他机制,如刺激血管平滑肌细胞增殖和血栓形成。国内有研究报道,仅给予新西兰大白兔高蛋氨酸饮食8周,即可形成兔AS 模型12。国外的研究则认为,单独给予新西兰大白兔高蛋氨酸饮食22周,能形成AS 模型13,14。本研究观察到给予20周高蛋氨酸饮食后,兔血浆Hcy 水平明显升高,同时伴有血脂明显升高,模型组可见腹主动脉狭窄及粥样硬化。Ste 2v

12、en 等15认为高蛋氨酸饮食能导致猴高Hcy血症,但血脂变化不明显。也有研究表明高蛋氨酸饮食不仅能导致兔高Hcy 血症,而且能导致高脂血症16。同时,高Hcy 多影响LDL 的代谢,但本研究中兔血清HDL 水平也增高,分析原因可能与实验动物及喂养时间不同有关。长期高水平的Hcy 刺激导致肝脏脂代谢紊乱,可能导致高脂血症,具体机制有待进一步探讨。给予普罗布考治疗后血浆Hcy 水平无下降,但血脂明显下降,同时主动脉组织形态明显改善,可见普罗布考改善血管病变与降低血脂有关,与血浆Hcy 水平无关。N F 2B 是Sen 等17从B 淋巴细胞核提取物中检测到的一种核蛋白因子,与多种心血管疾病及AS 有

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