川芎对心肌缺血再灌注损伤大鼠的影响(1)
[摘要]目的 探讨川芎对心肌缺血再灌注损伤大鼠心肌梗死及心肌NF-κB的影响。方法 将雄性SD大鼠30只随机分为假手术组、对照组、川芎组，采用结扎左冠状动脉前降支法制备心肌缺血再灌注损伤模型。记录对照组、川芎组心肌缺血、梗死面积，采用免疫组化方法观察各组心肌组织细胞中NF-κB蛋白表达，并比较血清心肌酶谱的变化。结果 ①川芎组大鼠心肌缺血、心肌梗死面积均明显小于对照组，差异有统计学意义(P<0.05)。②与假手术组比较，对照组血CK-MB、LDH含量明显升高，差异有统计学意义(P<0.05)；川芎组血CK-MB、LDH含量明显低于对照组，差异有统计学意义(P<0.05)；对照组NF-κB光密度值明显高于假手术组和川芎组，川芎组略高于假手术组，差异有统计学意义(P<0.05)。结论 川芎有利于减小心肌缺血再灌注损伤大鼠心肌缺血、梗死的面积，抑制心肌NF-κB的活性。
[关键词]川芎；缺血再灌注损伤；核因子-κB；心肌梗死
[中图分类号] R363 [文献标识码] A [文章编号] 1674-4721(2016)06(c)-0109-03
[Abstract]Objective To investigate the effect of Chuanxiong on yocardial infarction and myocardial NF-κB of rats with myocardial ischemia-reperfusion injury.Methods 30 male SD rats were randomly divided into sham group，the control group，Chuanxiong group.The myocardial ischemia-reperfusion injury model was made by ligation of the left anterior descending coronary artery.The myocardial ischemia and infarction area in the control group and Chuanxiong group were recorded，the NF-κB protein expression of myocardial cells was observed by immunohistochemical method，and serum myocardial enzyme change was compared.Results ①The myocardial ischemia and infarction area in the Chuanxiong group was higher than that of the control group，with statistical difference(P<0.05).②The level of blood CK-MB，LDH in the control group was significantly increased respectively compared with the sham group，with statistical difference(P<0.05).The level of blood CK-MB， LDH in Chuanxiong group was significantly lower than that of the control group respectively，with statistical difference(P<0.05).The NF-κB in the control group was significantly higher than that of the sham group and Chuanxiong group，Chuanxiong group slightly higher than that of the sham group，with statistical difference(P<0.05).Conclusion Chuanxiong can reduce myocardial ischemia and infarct size and inhibit myocardial NF-κB activity in rats with myocardial ischemia-reperfusion injury.
[Key words]Chuanxiong；Ischemia-reperfusion injury；Nuclear factor-κB；Miocardial infarction
冠心病是一种严重危害人类健康及生命的心血管疾病，其发病率和病死率有逐年升高的趋势。虽然近年来随着经皮冠状动脉介入和冠状动脉旁路移植术等治疗方法的广泛开展，冠心病的治愈率和生存率较以往有了较大提高，但是心肌缺血再灌注损伤的发生率仍然居高不下[1-2]。心肌缺血再灌注损伤发生后可诱导核因子-κB(NF-κB)的激活，后者可通过调控多种基因的表达参与缺血后损伤反应，因而NF-κB在心肌缺血再灌注中具有重要作用[3]。近年来，不断有研究指出，药物可以缓解缺血再灌注动物模型的心肌损伤[4]。临床中应用川芎治疗心肌缺血及再灌注损伤取得了较好效果，本研究旨在探讨川芎对心肌缺血再灌注损伤大鼠的影响。
1资料与方法
1.1 药物制备
取川芎30 g加10倍量水置容器中并浸泡20 min，加热煮沸后改用文火煎煮30 min，滤过后的滤渣再加8倍量水煎煮30 min后滤过，合并两次滤液于烧杯中备用。
1.2 动物模型制备及分组
取健康雄性SD大鼠(广州医学院实验动物中心)30只，将其随机分为三组，每组10只，其中假手术组(仅穿线，不结扎左冠状动脉前降支，生理盐水20 ml/kg)、对照组(生理盐水20 ml/kg)、川芎组(川芎50 mg/kg，川芎由惠州市中医医院中药房提供)每天灌胃给相应药物1次，持续2周。制备方法：3%戊巴比妥钠腹腔注射麻醉后，大鼠四肢固定于手术台上，将注射器针头置于大鼠四肢皮下接心电图机，记录Ⅱ导联心电图。切口选择颈前正中位置，将皮肤、筋膜切开，并将其逐层分离直至气管，将气管切开并插入自制气管插管，连接小型动物呼吸机，待大鼠呼吸抵抗消失、呼吸平稳后开胸。将左侧皮肤剪开并逐层分离，在4～5助骨间进行钝性分离，将肋骨撑开后打开心包，暴露心脏，沿室间沟仔细找寻左冠状动脉前降支，在左心耳与肺动脉圆锥处将管结扎，随着缺血时间延长心尖处可见心肌发白、水肿明显，心尖搏动减弱表现；观察心电图，可出现ST段抬高表现；缺血30 min后，将结扎线放开，恢复血流灌注，如缺血心肌恢复为正常心肌色，Ⅱ导联心电图出现ST段逐渐回落，则证明心肌缺血再灌注模型制备成功，再灌注120 min后，摘除大鼠心肌及并取血待测。 (朱俊新 饶李亮)
[关键词]川芎；缺血再灌注损伤；核因子-κB；心肌梗死
[中图分类号] R363 [文献标识码] A [文章编号] 1674-4721(2016)06(c)-0109-03
[Abstract]Objective To investigate the effect of Chuanxiong on yocardial infarction and myocardial NF-κB of rats with myocardial ischemia-reperfusion injury.Methods 30 male SD rats were randomly divided into sham group，the control group，Chuanxiong group.The myocardial ischemia-reperfusion injury model was made by ligation of the left anterior descending coronary artery.The myocardial ischemia and infarction area in the control group and Chuanxiong group were recorded，the NF-κB protein expression of myocardial cells was observed by immunohistochemical method，and serum myocardial enzyme change was compared.Results ①The myocardial ischemia and infarction area in the Chuanxiong group was higher than that of the control group，with statistical difference(P<0.05).②The level of blood CK-MB，LDH in the control group was significantly increased respectively compared with the sham group，with statistical difference(P<0.05).The level of blood CK-MB， LDH in Chuanxiong group was significantly lower than that of the control group respectively，with statistical difference(P<0.05).The NF-κB in the control group was significantly higher than that of the sham group and Chuanxiong group，Chuanxiong group slightly higher than that of the sham group，with statistical difference(P<0.05).Conclusion Chuanxiong can reduce myocardial ischemia and infarct size and inhibit myocardial NF-κB activity in rats with myocardial ischemia-reperfusion injury.
[Key words]Chuanxiong；Ischemia-reperfusion injury；Nuclear factor-κB；Miocardial infarction
冠心病是一种严重危害人类健康及生命的心血管疾病，其发病率和病死率有逐年升高的趋势。虽然近年来随着经皮冠状动脉介入和冠状动脉旁路移植术等治疗方法的广泛开展，冠心病的治愈率和生存率较以往有了较大提高，但是心肌缺血再灌注损伤的发生率仍然居高不下[1-2]。心肌缺血再灌注损伤发生后可诱导核因子-κB(NF-κB)的激活，后者可通过调控多种基因的表达参与缺血后损伤反应，因而NF-κB在心肌缺血再灌注中具有重要作用[3]。近年来，不断有研究指出，药物可以缓解缺血再灌注动物模型的心肌损伤[4]。临床中应用川芎治疗心肌缺血及再灌注损伤取得了较好效果，本研究旨在探讨川芎对心肌缺血再灌注损伤大鼠的影响。
1资料与方法
1.1 药物制备
取川芎30 g加10倍量水置容器中并浸泡20 min，加热煮沸后改用文火煎煮30 min，滤过后的滤渣再加8倍量水煎煮30 min后滤过，合并两次滤液于烧杯中备用。
1.2 动物模型制备及分组
取健康雄性SD大鼠(广州医学院实验动物中心)30只，将其随机分为三组，每组10只，其中假手术组(仅穿线，不结扎左冠状动脉前降支，生理盐水20 ml/kg)、对照组(生理盐水20 ml/kg)、川芎组(川芎50 mg/kg，川芎由惠州市中医医院中药房提供)每天灌胃给相应药物1次，持续2周。制备方法：3%戊巴比妥钠腹腔注射麻醉后，大鼠四肢固定于手术台上，将注射器针头置于大鼠四肢皮下接心电图机，记录Ⅱ导联心电图。切口选择颈前正中位置，将皮肤、筋膜切开，并将其逐层分离直至气管，将气管切开并插入自制气管插管，连接小型动物呼吸机，待大鼠呼吸抵抗消失、呼吸平稳后开胸。将左侧皮肤剪开并逐层分离，在4～5助骨间进行钝性分离，将肋骨撑开后打开心包，暴露心脏，沿室间沟仔细找寻左冠状动脉前降支，在左心耳与肺动脉圆锥处将管结扎，随着缺血时间延长心尖处可见心肌发白、水肿明显，心尖搏动减弱表现；观察心电图，可出现ST段抬高表现；缺血30 min后，将结扎线放开，恢复血流灌注，如缺血心肌恢复为正常心肌色，Ⅱ导联心电图出现ST段逐渐回落，则证明心肌缺血再灌注模型制备成功，再灌注120 min后，摘除大鼠心肌及并取血待测。 (朱俊新 饶李亮)