粉防己碱预处理对心肌缺血/再灌注损伤大鼠肿瘤坏死因子-α表达及核因子活化的影响

The Effect of Tetrandrine on the Expression of Tumor Necrosis Factor-α and the Activation of Nuclear Factor-κB Against Myocardial Ischemia/reperfusion Injury in Rats

目的探讨粉防己碱预处理对心肌缺血/再灌注过程中大鼠肿瘤坏死因子-α表达及与核因子活化的影响以及相关关系。方法60只SD大鼠随机分为3组:缺血/再灌注损伤组、假手术组、粉防己碱治疗组,缺血/再灌注损伤组结扎大鼠左前降支造成心肌缺血30min后再灌注24h后处死大鼠;假手术组只穿针不结扎,余步骤同缺血/再灌注损伤组;粉防己碱治疗组在缺血前30min腹腔注射粉防己碱,余步骤同缺血/再灌注损伤组。24h后取心肌标本,用酶联免疫吸附法检测心肌组织中肿瘤坏死因子-α表达水平,凝胶电泳迁移率变化分析检测核因子的活性。结果粉防己碱组与缺血再灌注组相比肿瘤坏死因子-α表达水平明显减低(208.40±25.12 VS 306.65±17.78,P<0.01)而与假手术组相比表达明显增强(208.40±25.12 VS 61.45±9.20,P<0.01)。粉防己碱组核因子的活性明显低于缺血再灌注组(29.58±1.56 VS 40.33±4.39,P<0.01)而与假手术组无显著性差异(29.58±1.56 VS 30.09±3.46,P>0.05)。结论粉防己碱预处理可以使心肌缺血/再灌注损伤过程中肿瘤坏死因子-α生成明显减少,核因子的活化受到有效抑制,从而起到减轻心肌缺血/再灌注损伤的作用。

Objective To investigate the effect of preconditioning tetrandrine on the expression of TNF-α and the activation of NF-κB and the relationship between them during rat myocardial reperfusion period. Methods 60 male Sprague-Dawley rats were randomly divided into 3 groups： Sham group, ischemia/reperfusion （I/R） group, tetrandrine group. The Sprague-Dawley rat underwent 30 min of left anterior descending （LAD） coronary occlusion and 24 - hour reperfusion to make ischemia/reperfusion （I/R） injury model in vivo. Sham group were not subjected to occlusion of artery. Tetrandrine group were injected tetrandrine to abdominal cavity before ischemia starting. The other procedures were the same as the ischemia/reperfusion group. Samples were collected 24h after reperfusion. The expression level of TNF-α protein was detected by ELISA. Electrephoretic mobility shift assay （EMSA） is used to analyze the binding activity of NF-κB. Results In tetrandrine group, the expression of TNF-α is significantly lower compared with ischemia/reperfusion group（ P 〈 0.01） and significandy higher than that of sham group（ P 〈 0.01）. In tetrandrine group, the activity of NF-κB is significandy lower than that of ischemic/reperfusion group（P 〈0.01） but there is no obvious difference between tetrandrine group and shame group（P 〉 0. 05）. Condusion Tetrandrine can extenuate myoca-rdial ischemia/reperfusion injury. This pharmacologic action is achiered through inhibiting the activation of NF-κB and decreasing the expression of TNF-α.