| | | 高同型半胱氨酸血症对球囊损伤大鼠颈动脉核转录因子NF
| | 论文下载 作者:徐会圃,刘长梅,冯义柏 【关键词】 ,甲硫氨酸 Effect of homocysteine on expressions of NFκB and protooncogene in ballooninjured rat carotid arteries 【Abstract】 AIM: To study the effect of homocysteine (Hcy) on the expressions of nuclear factorκB (NFκB) P65 protein and cfos, cjun mRNA in common carotid artery of rats after balloon injury and to explore the possible mechanism by which homocysteine exacerbates neointimal hyperplasia after angioplasty. METHODS: The expressions of NFκB P65 protein and cfos and cjun mRNA were detected by Western blot and semiquantitative RTPCR respectively in the common carotid artery. RESULTS: The expressions of NFκB P65 protein and cfos and cjun mRNA were significantly higher in low methionine (LM) group(1.12±0.13, 1.40±0.21, 1.43±0.25)(P<0.05) and high methionine (HM) group(1.50±0.37, 1.68±0.27, 1.71±0.30)(P<0.01)than those in control group(0.64±0.06, 1.10±0.15, 1.00±0.13). The expressions of NFκB P65 protein and cfos, cjun mRNA were also significant higher in HM group than those in LM group (P<0.05). CONCLUSION: Homocysteine may upregulate the transcription of cfos and cjun mRNA in common carotid artery after balloon angioplasty by NFκB pathway, which may be responsible for neointimal hyperplasia induced by homocysteine after angioplasty. 【Keywords】 methionine; homocysteine; balloon injury; NFkappaB; protooncogenes 【摘要】 目的论文下载: 观察高蛋氨酸饮食诱导的论文下载高同型半胱氨酸血症对大鼠颈动脉球囊损伤后血管组织NFκBP65蛋白和原癌基因cfos及cjun mRNA表达的影响,探讨同型半胱氨酸在血管球囊损伤后新内膜过度增生中的可能机制. 方法: 采用Western blot及RTPCR方法分别检测血管组织NFκBP65蛋白和cfos及cjun mRNA表达,并做半定量分析. 结果: 低、高蛋氨酸组血管组织NFκBP65蛋白、cfos及cjun mRNA的表达均高于对照组,分别为1.12±0.13 vs 0.64±0.06 (P<0.05), 1.50±0.37 vs 0.64±0.06 (P<0.01), 1.40±0.21 vs 1.10±0.15 (P<0.05), 1.43±0.25 vs 1.10±0.15 (P<0.01)及1.68±0.27 vs 1.00±0.13 (P<0.05), 1.71±0.30 vs 1.00±0.13 (P<0.01),高蛋氨酸组也明显高于低蛋氨酸组(P<0.05). 结论: 同型半胱氨酸有可能通过NFκB通路上调血管内皮损伤后动脉组织原癌基因cfos及cjun mRNA的表达,这可能是其促进血管内皮球囊损伤后新内膜过度增生的机制之一. 【关键词】 甲硫氨酸;同型半胱氨酸;球囊损伤;原癌基因;NFκB 0引言 实验研究和临床观察表明,血中同型半胱氨酸(homocysteine, Hcy)升高可引起动脉粥样硬化和血栓形成[1-2]. 然而其在血管内皮球囊损伤后新内膜增生中的作用及其机制尚不清楚. 我们用高蛋氨酸(methionine)饮食诱导高同型半胱氨酸血症,观察其对血管内皮球囊损伤后血管组织NFκB P65蛋白及原癌基因表达的影响,以探讨Hcy促进血管内皮球囊损伤后新内膜过度增生的可能机制. 1材料和方法 1.1材料 健康雄性12 wk SD大鼠30只(华中科技大学同济医学院动物中心提供),体质量280~320 g,随机分为对照组(C)(给普通饲料),低蛋氨酸组(LM)(含蛋氨酸12 g/kg饮食),高蛋氨酸组(HM)(含蛋氨酸20 g/kg饮食),每组10只,各组均自由进食. 蛋氨酸(北京中山生物技术有限公司,化学纯,批号: 1903001),总RNA提取试剂(Trizol)及逆转录试剂盒为Giblo公司产品,TaqDNA聚合酶cfos, cjun及内参βactin上、下游引物为TaKaRa大连宝生物工程有限公司产品;兔抗大鼠NFκB P65多克隆抗体、Western blot检测试剂盒均购自武汉博士德公司. 自制2F球囊导管购自天津乳胶制品厂;PCR扩增仪(美国MJPC150公司),图像分析仪为上海四星公司产品. 高效液相色谱仪(美国water公司).
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