普羅布考對高同型半胱氨酸血癥致兔動脈粥樣硬化的作用_圖文

1、論著普羅布考對高同型半胱氨酸血癥致兔動脈粥樣硬化的作用吳樂a #,黎紅華a ,武強a ,王曉昆b廣州軍區(qū)武漢總醫(yī)院a.神經內科,b.醫(yī)學實驗中心,武漢430070【收稿日期】2010207207#【摘要】目的:觀察普羅布考對高同型半胱氨酸(Hcy 血癥致兔動脈粥樣硬化(AS 的作用。方法:將24只大白兔分為對照組、模型組和治療組各8只,對照組給予基礎飼料喂養(yǎng),模型組和治療組均在基礎飼料中加入1%的蛋氨酸,此外治療組同時用普羅布考灌胃(1g/d ;飲食量200g/d ,均喂養(yǎng)20周。分別于第20周末檢測血總同型半胱氨酸(t Hcy 、總膽固醇(TC 、甘油三酯(T G 、高密度脂蛋白膽固醇(HD

2、L 2C 和低密度脂蛋白膽固醇(LDL 2C 。第20周末處死動物,剝離腹主動脈,用Western blot 檢測血管中核轉錄因子2B (N F 2B 蛋白表達,光鏡下觀察主動脈病理形態(tài)學變化。結果:治療組血中TC 、T G 、HDL 2C 和LDL 2C 水平低于模型組(P 0.05,t Hcy水平無統(tǒng)計學差異;治療組主動脈N F 2B 蛋白表達水平明顯低于模型組(P 0.05;主動脈病理形態(tài)學檢查顯示治療組病變輕于模型組。結論:普羅布考能降低血脂,下調N F 2B ,減輕內膜增生程度,具有抗AS 的作用?！娟P鍵詞】普羅布考;同型半胱氨酸;動脈粥樣硬化;核轉錄因子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動脈粥樣硬化(at hero sclero sis ,AS 是一種慢性炎癥反應過程,主要累及中等以上的肌性動脈。核轉錄因子2B (nuclear fac 2tor 2B ,N F 2B 的激活是

8、AS 形成的重要環(huán)節(jié)。同型半胱氨酸(homocysterine ,Hcy 是導致AS的獨立危險因素1,2。普羅布考(p robuco1已作為降脂藥應用于臨床,同時具有抗AS作用,推測可能與改善內皮功能有關3。本研究以高蛋氨酸飲食誘導兔高Hcy血癥及AS模型,探討普羅布考在高Hcy血癥致AS過程中的作用。1材料與方法1.1材料動物:雄性純種新西蘭大白兔24只,體質量2.02.5kg,購自湖北省動物實驗中心。主要試劑與材料:兔抗N F2B p65(購于美國Santa Cruz公司,辣根過氧化物酶標記的二抗(羊抗兔和羊抗鼠(購于武漢Bo ster公司,普羅布考粉劑(購于山東齊魯制藥有限公司。1.2方

9、法2結果治療組較模型組血清血脂水平明顯下降,但仍高于對照組,有統(tǒng)計學差異(P 0.05,模型組和治療組血漿Hcy水平差異無統(tǒng)計學意義,見表1。Western blot顯示,模型組N F2B p65蛋白表達較對照組顯著增高(P=0.009。治療組N F2B p65蛋白表達降低,與模型組相比差異有統(tǒng)計學意義(P=0.046,仍高于對照組(P=0.027,見圖1A-B。H E染色顯示對照組血管壁內膜光滑完整,中膜平滑肌細胞排列規(guī)則;模型組血管內膜明顯增厚,可見一處內膜下出血(箭頭處,中膜平滑肌細胞排列紊亂;治療組血管內膜增厚明顯減輕,中膜平滑肌細胞排列規(guī)則,見圖2A-C。3討論Schwartz等4研

10、究發(fā)現,普羅布考治療后HDL顆粒的直徑變小,數目不變,普羅布考治療使HDL2C失去膽固醇部分而使單位體積血液中HDL2C的總重量降低。由于普羅布考在降膽固醇的同時降低HDL2C,而淡出市場。然而,臨床研究發(fā)現,普羅布考雖然降低HDL2C,但顯示出強大的抗AS、抑制再狹窄和改善血管舒張功能的作用5,6。表13組血脂及Hcy 水平比較(x -s 組別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與對照組比較,P 0.05,P 0.01;與模型

11、組比較,P 0.05 血漿Hcy 水平嚴重升高的患者常伴早發(fā)性AS 。血漿高Hcy 引起心血管疾病的機制未完全明了,目前認為主要與以下幾個方面有關:導致內皮細胞損傷及功能障礙7,8;促進脂蛋白過氧化9;引起許多炎癥介質釋放10;增加肝細胞TC 合成等11;其他機制,如刺激血管平滑肌細胞增殖和血栓形成。國內有研究報道,僅給予新西蘭大白兔高蛋氨酸飲食8周,即可形成兔AS 模型12。國外的研究則認為,單獨給予新西蘭大白兔高蛋氨酸飲食22周,能形成AS 模型13,14。本研究觀察到給予20周高蛋氨酸飲食后,兔血漿Hcy 水平明顯升高,同時伴有血脂明顯升高,模型組可見腹主動脈狹窄及粥樣硬化。Ste 2v

12、en 等15認為高蛋氨酸飲食能導致猴高Hcy血癥,但血脂變化不明顯。也有研究表明高蛋氨酸飲食不僅能導致兔高Hcy 血癥,而且能導致高脂血癥16。同時,高Hcy 多影響LDL 的代謝,但本研究中兔血清HDL 水平也增高,分析原因可能與實驗動物及喂養(yǎng)時間不同有關。長期高水平的Hcy 刺激導致肝臟脂代謝紊亂,可能導致高脂血癥,具體機制有待進一步探討。給予普羅布考治療后血漿Hcy 水平無下降,但血脂明顯下降,同時主動脈組織形態(tài)明顯改善,可見普羅布考改善血管病變與降低血脂有關,與血漿Hcy 水平無關。N F 2B 是Sen 等17從B 淋巴細胞核提取物中檢測到的一種核蛋白因子,與多種心血管疾病及AS 有

13、關18,19。本研究觀察到給予高蛋氨酸飲食后兔主動脈N F 2B 表達水平明顯升高,給予普羅布考治療后N F 2B 表達水平明顯下降,同時主動脈組織形態(tài)明顯改善,可見普羅布考可能通過調節(jié)N F 2B 表達水平而改善血管病變?！緟⒖嘉墨I】1Huang W ,Glass C K.Nuclear receptors andinflammation cont rol :molecular mechanisms and pat hop hysiological relevance J .Arte 2rioscler Thromb Vasc Biol ,2010,30(8:1542-1549.2Lauer

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