Dai Qiu-Fu, Gao Jun-Hong, Xin Juan-Juan, Liu Qun, Jing Xiang-Hong, Yu Xiao-Chun
Acupuncture and Moxibustion Department, Beijing Hospital of Traditional Chinese Medicine Affiliated to Capital Medical University, Capital Medical University, Beijing Key Laboratory of Acupuncture Neuromodulation, 23 Art Museum Back Street, Beijing 100700, China.
Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences, 16 Nanxiaojie, Dongzhimennei, Beijing 100700, China.
Evid Based Complement Alternat Med. 2019 Dec 2;2019:6721286. doi: 10.1155/2019/6721286. eCollection 2019.
To investigate the roles played by A2b receptor and the key Ca signaling components in the mediation of the cardioprotection of electroacupuncture pretreatment in the rats subjected to myocardial ischemia and reperfusion.
SD rats were randomly divided into a normal control (NC) group, ischemia/reperfusion model (M) group, electroacupuncture pretreatment (EA) group, and electroacupuncture pretreatment plus A2b antagonist (EAG) group. The ischemia/reperfusion model was made by ligation and loosening of the left descending branch of the coronary artery in all groups except the NC group. The EA group was pretreated with electroacupuncture at the (PC6) point once a day for three consecutive days before the modeling. The elevation of the ST segment, arrhythmia scores, and myocardial infarction size of each group was measured. The relative expression levels of A2b, RyR2, SERCA2a, NCX1, P-PLB(S16)/PLB, and Troponin C/Troponin I proteins in the injured myocardium were detected by multiple fluorescence western blot.
The level of ST segment, arrhythmia scores, and infarct size in the M group was significantly higher/larger than that in the NC group after ischemia and reperfusion, while all the three indices mentioned above in the EA group were significantly lower/smaller than those in the M group after reperfusion. The expression of the proteins of adenosine receptor 2b(A2b), ryanodine receptor 2(RyR2), and sarco(endo)plasmic reticulum Ca-ATPase 2a (SERCA2a) in the EA group was significantly enhanced as compared with the M group, while in the EAG group, the contents of A2b were significantly lower than those in the EA group, and RyR2 was higher in the EAG group. In comparison with the NC group, the relative expression of NCX1 protein in M, EA, and EAG groups was not changed significantly. The ratio of phosphorylated phospholamban (P-PLB) over phospholamban (PLB) in the M group was significantly lower than that in the NC group, and the ratio in the EA group was significantly increased as compared with the M group, while the ratio of Troponin C/Troponin I in the EA group was significantly decreased in comparison with that in other groups.
Electroacupuncture pretreatment could reduce ischemia and reperfusion-induced myocardial injury via possibly increasing the A2b content and regulating the key Ca signaling components, namely inhibiting RyR2 and enhancing P-PLB(S16)/PLB ratio and SERCA2a proteins, so as to diminish the intracellular Ca overload and consequently lessen the myocardial injury.
探讨A2b受体及关键钙信号成分在电针预处理介导的心肌缺血再灌注大鼠心脏保护作用中所起的作用。
将SD大鼠随机分为正常对照组(NC组)、缺血/再灌注模型组(M组)、电针预处理组(EA组)和电针预处理加A2b拮抗剂组(EAG组)。除NC组外,其余各组均通过结扎和松开冠状动脉左前降支制备缺血/再灌注模型。EA组在建模前连续3天每天于内关穴(PC6)进行一次电针预处理。测量各组ST段抬高、心律失常评分及心肌梗死面积。采用多荧光western blot法检测损伤心肌中A2b、RyR2、SERCA2a、NCX1、P-PLB(S16)/PLB和肌钙蛋白C/肌钙蛋白I蛋白的相对表达水平。
缺血再灌注后,M组的ST段水平、心律失常评分及梗死面积显著高于NC组,而EA组再灌注后的上述三项指标均显著低于M组。与M组相比,EA组腺苷受体2b(A2b)、兰尼碱受体2(RyR2)和肌浆网钙ATP酶2a(SERCA2a)蛋白的表达显著增强,而在EAG组,A2b含量显著低于EA组,RyR2则高于EA组。与NC组相比,M组、EA组和EAG组中NCX1蛋白的相对表达无明显变化。M组磷酸化受磷蛋白(P-PLB)与受磷蛋白(PLB)的比值显著低于NC组,与M组相比,EA组该比值显著升高,而与其他组相比,EA组肌钙蛋白C/肌钙蛋白I的比值显著降低。
电针预处理可能通过增加A2b含量并调节关键钙信号成分,即抑制RyR2并提高P-PLB(S16)/PLB比值和SERCA2a蛋白水平,从而减少细胞内钙超载,进而减轻心肌损伤,降低缺血再灌注诱导的心肌损伤。