参附注射液对大鼠急性缺血/再灌注损伤心肌核转录因子-κB的影响

Effects of shenfu injection on nuclear factor-κB during myocardial ischemia/reperfusion injury in rats

目的:探讨参附注射液保护大鼠心肌缺血 再灌注损伤作用与机理。方法:采用大鼠心脏左冠状动脉前降支缺血 再灌注模型,缺血30min ,再灌注6 0min。2 4只SD大鼠随机分为3组:假手术组;缺血 再灌注组;参附注射液组。酶联免疫吸附实验法检测血浆中肿瘤坏死因子 -α(TNF- α)、白细胞介素- 6(IL- 6 )浓度,免疫印记法测心肌核转录因子- κB(NF -κB)活性水平,电镜观察心肌超微结构。结果:缺血 再灌注组心肌NF -κB的活性,血浆TNF- a、IL -6浓度较假手术组明显上升(P <0 .0 1)。参附注射液组中上述指标与缺血 再灌注组比较均显著下降(P <0 .0 1) ,电镜下参附注射液组中心肌超微结构较缺血再灌注组明显减轻并接近正常。结论:参附注射液通过抑制缺血心肌中NF- κB的活性,降低促炎因子水平,对心肌起保护作用。

AIM: To study the protective effects of shenfu injection on myocardial ischemia/reperfusion (I/R) injury in rats and its potential mechanisms. METHODS: Myocardial ischemia/reperfusion was produced by tying and untying of left anterior descending coronary artery. Ischemia lasted for 30 min and reperfusion for 60 min. Twenty-four healthy male SD rats weighing 230-280 g were randomly divided into three groups (n=8 in each): sham-operation group, I/R group, and shenfu group which the shenfu injection (10 ml·kg -1) were injected intraperitoneally 30 min before ischemia. The plasma concentration of tumor necrosis factor-α (TNF-α), interleukin-6(IL-6) were measured by ELISA. The heart was harvested and levels of the nuclear factor kappaB (NF-κB) activity were determined by Ecl-western blot analysis and ultrastructures were observed by electron microscopy. RESULTS: NF-κB binding activity in myocardial nuclear and the plasma concentration of IL-6, TNF-α were significantly increased in I/R group than that in the sham-operation group (P< 0.01), and they were markedly reduced in shenfu group compared with I/R group (P< 0.01). In addition, electron microscopic examination showed more serious injury of the myocardium ultrastructure in I/R group, while in shenfu group the myocardial ultrastructure could improve close to normal. CONCLUSION: Shenfu injection can inhibit NF-κB activity in postischemic myocardium and lead to down-regulation of proinflammtory cytokine expression, which may be one of molecular mechanisms of shenfu injection in cardioprotection.