电针预处理对急性心肌缺血小鼠心功能及免疫炎性反应的影响

Effects of electroacupuncture pretreatment on cardiac function and immune inflammatory response in mice with acute myocardial ischemia

目的:观察电针预处理对急性心肌缺血模型小鼠心脏射血分数(EF),脾脏、心脏巨噬细胞数量及心肌组织核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)、白细胞介素-1β(IL-1β)表达的影响,探讨电针预处理促心肌保护的可能机制。方法:30只雄性C57BL/6J小鼠随机分为对照组、模型组和电针预处理组,每组10只。模型组和电针预处理组通过结扎小鼠心脏冠状动脉左前降支建立急性心肌缺血模型,对照组仅在小鼠心脏冠状动脉左前降支穿线不做结扎。电针预处理组电针小鼠双侧"内关",疏密波,频率2 Hz/15 Hz,强度2 m A,每次电针20min,每天1次,共干预3d后造模。采用超声心动图M图EF值评价心功能;流式细胞计数法检测脾脏、心脏巨噬细胞数量;免疫印迹法检测小鼠心肌组织NLRP3、IL-1β蛋白表达水平。结果:与对照组比较,模型组小鼠EF值下降(P<0.001),心脏、脾脏巨噬细胞数量增加(P<0.001),心肌组织NLRP3、IL-1β蛋白表达水平上升(P<0.001,P<0.01);与模型组比较,电针预处理组小鼠EF值上升(P<0.01),心脏、脾脏巨噬细胞数量下降(P<0.01),心肌组织NLRP3、IL-1β蛋白表达水平下降(P<0.01,P<0.05)。结论:电针预处理可减少急性心肌缺血小鼠脾脏、心脏巨噬细胞数量,下调心肌组织NLRP3、IL-1β蛋白表达,减轻心肌局部炎性反应,从而实现心肌保护效应。

Objective To observe the effects of electroacupuncture(EA) pretreatment on the cardiac ejection fraction(EF), the number of macrophages in spleen and heart, and the expression of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3) and interleukin-1β(IL-1β) in myocardium in mice with acute myocardial ischemia, and to explore the possible mechanism of EA pretreatment on promoting myocardial protection. Methods A total of 30 male C57 BL/6 J mice were randomly divided into a control group, a model group and an EA pretreatment group, 10 rats in each group. The acute myocardial ischemia model was established by ligating the left anterior descending branch of the coronary artery in the model group and EA pretreatment group, while threading but no ligating at left anterior descending branch of the coronary artery was applied in the control group. In the EA pretreatment group, mice were intervented with EA at bilateral "Neiguan"(PC 6), disperse-dense wave, frequency of 2 Hz/15 Hz, intensity of 2 m A;each EA treatment last for 20 min, once a day, and 3-day treatment was given before model establishment. The EF value was evaluated by ultrasonic cardiogram;the number of macrophages in spleen and heart was measured by flow cytometry;the expression level of NLRP3 and IL-1β in myocardium was measured by Western blot. Results Compared with the control group, the EF value was decreased in the model group(P<0.001), the number of macrophages in the heart and spleen was increased(P<0.001), and the expression level of NLRP3 and IL-1β in the myocardium was increased(P<0.001, P<0.01). Compared with the model group, the EF value was increased in the EA pretreatment group(P<0.01), the number of macrophages in the heart and spleen was decreased(P<0.01), and the expression level of NLRP3 and IL-1β in the myocardium was decreased(P<0.01, P<0.05).Conclusion EA pretreatment could reduce the number of macrophages in spleen and heart, down-regulate the expression of NLRP3 and IL-1β in myocardial tissue in mice with acute myocardial ischemia, which could relieve the local inflammatory response and achieve the myocardial protective effect.