二氨基庚二酸激活NOD1信号通路对心肌缺血再灌注损伤的影响

Effect of DAP activated NOD1 signaling pathway on myocardial ischemia/reperfusion injury

目的:探讨二氨基庚二酸(DAP)激活核苷酸结合寡聚化结构域蛋白1(NOD1)信号通路对心肌缺血再灌注(I/R)损伤的影响。方法:将40只C57/BL6雄性小鼠随机分为4组,即假手术组、I/R组、DAP组和I/R+DAP组,每组10只。对I/R组和I/R+DAP组小鼠进行心肌缺血再灌注模型制备,假手术组进行相同的手术操作,但只穿线不结扎,DAP组和I/R+DAP组小鼠在冠状动脉结扎前30 min腹腔注射DAP,假手术组和I/R组小鼠注射等剂量的生理盐水;观察各组小鼠心肌梗死、心肌细胞凋亡、炎症反应及NF-κB p65磷酸化情况。结果:I/R组NOD1 mRNA和NOD1蛋白表达水平均显著高于假手术组(P<0.05);I/R组梗死面积比值明显高于假手术组,且I/R+DAP组小鼠梗死面积最高(P<0.05),而DAP组与假手术组差异无统计学意义;I/R组和I/R+DAP组TUNEL阳性细胞数显着高于假手术组,I/R+DAP组阳性细胞数最多(P<0.05),而DAP组与假手术组差异无统计学意义;I/R组和I/R+DAP组炎症细胞浸润数明显多于假手术组,I/R+DAP组小鼠最高(P<0.05);I/R组和I/R+DAP组磷酸化的NF-κB p65蛋白水平显著高于假手术组,且I/R+DAP组最高(P<0.05)。结论:DAP激活NOD1可加重缺血再灌注损伤后小鼠心脏心肌细胞的梗死面积,增加心肌细胞的凋亡和炎症反应,该机制与NF-κB信号通路的激活有关。

Objective： To investigate the effect of diaminopimelate（ DAP） activated NOD1 signaling pathway on myocardial ischemia/reperfusion（ I/R） injury. Methods： 40 C57/BL6 male mice were randomly divided into four groups（ n = 10） ： sham group,I/R group,DAP group and I/R ＋ DAP group. Mice in I/R group and I/R ＋ DAP group were treated with myocardial ischemia-reperfusion model,sham group was treated with the same operation,but only threaded without deligation. DAP group and I/R ＋ DAP group were treated with intraperitoneal injection of DAP at 30 min before coronary artery ligation,sham operation group and I/R group,mice in sham group and I/R group were injected with normal saline. Myocardial infarction,myocardial apoptosis,inflammatory reaction and NF-κB p65 phosphorylation were observed in each group. Results： The expression of NOD1 mRNA and NOD1 protein in I/R group were significantly higher than those in sham operation group（ P〈0. 05）. The infarct size in I/R group was significantly higher than that in sham operation group,and the infarct size of I/R ＋ DAP group was the highest（ P〈0. 05）. The number of TUNEL positive cells in I/R group and I/R ＋ DAP group were significantly higher than that in sham group,and the number of positive cells in I/R ＋ DAP group was the highest（ P〈0. 05）.The infiltration rate of inflammatory cells in I/R group and I/R ＋ DAP group was significantly higher than that in sham group,and I/R ＋ DAP group were the highest（ P〈0. 05）. The levels of phosphorylated NF-κB p65 protein in I/R group and I/R ＋ DAP group were significantly higher than those in the sham operation group,and I/R ＋ DAP group were the highest（ P〈0. 05）. Conclusion： This study demonstrates that activation of NOD1 through DAP can aggravate the infarct size of cardiac cardiomyocytes and increase the apoptosis and inflammatory response of cardiomyocytes in mice after ischemia/reperfusion injury,and which is related to the activation of NF-κB signaling pathway.