【作者】 赵胜男；

【导师】 胡治平；

【作者基本信息】 中南大学 ， 临床医学， 2012， 博士

【摘要】 目的观察磷酸吡哆醛丁咯地尔(buflomedil pyridoxal phosphate, BPP)对大鼠高同型半肤氨酸血症生物学效应的影响,探讨磷酸吡哆醛丁咯地尔能否拮抗高同型半肤氨酸血症的生物学效应及其作用机制。方法健康雄性SD大鼠40只,随机分为5组,每组8只：1组为BPP组,给予2%蛋氨酸饲料(20g蛋氨酸/1kg正常饲料)+BPP灌胃；2组为丁咯地尔+维生素B6组,给予2%蛋氨酸饲料+丁咯地尔、维生素B6灌胃；3组为丁咯地尔组,给予2%蛋氨酸饲料+丁咯地尔灌胃；4组为蛋氨酸模型组,给予2%蛋氨酸饲料+生理盐水灌胃；5组为正常对照组,给予正常饲料+生理盐水灌胃。药物灌胃剂量根据等效剂量系数折算法换算,每只大鼠每天灌胃两次,连续喂养8周。第8周末,用10%水合氯醛注射液麻醉大鼠,腹主动脉取血离心制备血浆,放入-20℃冻存,用ELISA检测同型半胱氨酸(homocysteine,Hcy)、超氧化物歧化酶(superoxide dismutase, SOD)的含量。摘取一侧颈动脉,用western blot检测劲动脉内皮型一氧化氮合成酶(endothelial nitric oxide synthase, eNOS)的蛋白表达。结果1.蛋氨酸模型组大鼠血浆Hcy浓度比正常对照组升高(P<0.01),BPP、丁咯地尔+维生素B6、丁咯地尔组血浆Hcy浓度均比蛋氨酸模型组低(P<0.05),BPP组血浆Hcy浓度低于丁咯地尔+维生素B6组、丁咯地尔组(P<0.01),接近正常对照组(P>0.05)2.蛋氨酸模型组大鼠血浆SOD浓度比正常对照组降低(P<0.01),BPP、丁咯地尔+维生素B6、丁咯地尔组血浆SOD浓度均比蛋氨酸模型组高(P<0.05),BPP组血浆SOD浓度高于丁咯地尔组(P<0.01),但与丁咯地尔+维生素B6组无明显差别(P>0.05),均低于正常对照组(P<0.05)。3.蛋氨酸模型组大鼠颈动脉eNOS蛋白表达比正常对照组降低(P<0.01),BPP、丁咯地尔+维生素B6、丁咯地尔组eNOS蛋白表达均比蛋氨酸模型组高(P<0.05),BPP组eONS蛋白表达高于丁咯地尔+维生素B6组、丁咯地尔组(P<0.01),接近正常对照组(P>0.05)4.所有大鼠颈动脉eONS蛋白表达与血浆Hcy浓度负相关,相关系数为-0.9,P<0.05。结论1.磷酸吡哆醛丁咯地尔、丁咯地尔+维生素B6、丁咯地尔均可以降低蛋氨酸负荷所致的高Hcy血症,并可能通过抑制Hcy诱导的SOD、eNOS表达下调来拮抗高Hcy血症的生物学效应。2.高浓度同型半胱氨酸可以通过抑制eNOS表达,减少NO生成。3.磷酸吡哆醛丁咯地尔拮抗高Hcy血症生物学效应的作用显著,优于丁咯地尔+维生素B6,可能成为防治高Hcy血症的新思路。更多还原

【Abstract】 ObjectiveTo evaluate the effect of buflomedil pyridoxal phosphate on hyperhomocysteinemia(HHcy).Methods40male Sprague-Dawley rats were randomly divided into five groups:buflomedil pyridoxal phosphate (BPP) group; buflomedil+VitB6group; buflomedil group; Met group; the controlled group. At the end of the8th weeks, get blood plasma and unilateral carotid artery to measure the concentration of homocysteine and superoxide dismutase, the protein expression of endothelial nitric oxide synthase.Results1.The Hcy concentration of Met group was significantly higher than controlled group(P<0.01), the Hcy concentration of the BPP, buflomedil+vitamin B6, buflomedil group was lower than Met group(P<0.01), the Hcy concentration of BPP group was significantly lower than buflomedil+vitamin B6group(P<0.01), close to the controlled group(P>0.05).2.The SOD concentration of Met group was significantly lower than controlled group(P<0.01), the SOD concentration of the BPP, buflomedil+vitamin B6, buflomedil group was higher than Met group(P <0.05); the SOD concentration of BPP group was close to buflomedil+vitamin B6group.3. The eNOS expression of Met group was significantly lower than controlled group(P<0.01), the eNOS expression of the BPP, buflomedil+vitamin B6, buflomedil group was higher than Met group(P<0.01), the eNOS expression of BPP group was significantly higher than buflomedil+vitamin B6group(P<0.01), close to the controlled group(P>0.05).4. The eNOS expression was negatively correlated with Hcy concentration.Conclusions1. Buflomedil pyridoxal phosphate, buflomedil+vitamin B6, buflomedil can all decrease plasma Hcy level, and may prevent the negative effect of Hcy to SOD, eNOS.2. Homocysteine can reduce NO production through suppressing the expression of eNOS.3. The influence of buflomedil pyridoxal phosphate on the biological effect of hyperhomocysteinemia was significant, and better than buflomedil+vitamin B6.更多还原