Objective: To investigate the mechanism of electroacupuncture (EA) with the involvement of sarcoplasmic reticulum Ca2+-ATPase2a (SERCA2a)/phospholamban (PLB) on the synergistic and attenuated effect of aconitine for heart failure.
Methods: Thirty SPF-ranked SD rats were randomly divided into a control group, a model group, an EA group, an aconitine group and an EA plus aconitine group, with 6 rats in each group. The rat model of acute heart failure was established by infusion of high-dose propranolol hydrochloride solution into the right femoral vein. After stabilized for 10 min in the modeled rats, EA was exerted at "Neiguan" (PC 6), with disperse-dense wave, 2 Hz/15 Hz in frequency, 3 mA in intensity, for 30 min in the EA group and the EA plus aconitine group; aconitine solution (10 μg/kg) was injected from the left femoral veins in the rats in the aconitine group and the EA plus aconitine group. Hemodynamic indexes such as the left ventricular systolic pressure (LVSP) and the maximum rate of increase/decrease of left ventricular pressure (±dp/dtmax) were detected and arrhythmia types were observed and scored. SERCA2a protein and PLB protein expressions in left ventricular myocardial tissue of rats were detected by multiplex fluorescence Western blot.
Results: Compared with the control group, LVSP and ±dp/dtmax all were decreased after modeling and at each time point after intervention in the model group (P<0.01). Compared with the model group, ±dp/dtmax was increased in the aconitine group and the EA group at 1 min after intervention (P<0.01, P<0.05), +dp/dtmax was increased at 10 to 60 min after intervention in the aconitine group and at 20 to 60 min after intervention in the EA group (P<0.01, P<0.05), LVSP was increased at 1 min after intervention in the EA group (P<0.01), while LVSP and ±dp/dtmax were all increased at 1 to 60 min after intervention in the EA plus aconitine group (P<0.01, P<0.05). Compared with the aconitine group, LVSP and +dp/dtmax were increased at 1 min after intervention in the EA group (P<0.01, P<0.05), LVSP and ±dp/dtmax at 1 min after intervention while +dp/dtmax at 20 to 60 min after intervention were all increased in the EA plus aconitine group (P<0.01, P<0.05). Compared with the EA group, +dp/dtmax was higher at 10 to 60 min after intervention in the EA plus aconitine group (P<0.01). Compared with the model group, arrhythmia score was higher in the aconitine group (P<0.01). Compared with the aconitine group, arrhythmia score was lower in the EA group and the EA plus aconitine group (P<0.01). As compared with the control group, the expression of SERCA2a protein in the left ventricular cardiomyocytes was decreased (P<0.01), while the expression of PLB protein was increased in the model group (P<0.01). Compared with the model group, the expression of SERCA2a protein was increased in both the EA group and the EA plus aconitine group (P<0.05, P<0.01), and PLB protein expression was decreased in each intervention group respectively (P<0.01, P<0.05). As compared with the EA group and the aconitine group, the expression of SERCA2a protein was increased and the expression of PLB protein was decreased in the EA plus aconitine group separately (P<0.05, P<0.01).
Conclusion: The intervention with electroacupuncture achieves the synergism/ attenuation effect of aconitine for the improvements in heart failure probably by up-regulating the expression of SERCA2a and down-regulating the expression of PLB in myocardial tissue.
目的：探讨肌浆网钙泵（SERCA2a）/磷酸受纳蛋白（PLB）在电针干预对乌头碱改善心力衰竭增效/减毒作用中的机制。方法：将30只SPF级SD大鼠随机分为对照组、模型组、电针组、乌头碱组和电针+乌头碱组，每组6只。采用右股静脉输注大剂量盐酸普萘洛尔溶液的方法制备大鼠急性心力衰竭模型。造模成功10 min后，电针组和电针+乌头碱组大鼠于“内关”行电针干预，予疏密波，频率2 Hz /15 Hz，电流强度3 mA，时间30 min；乌头碱组和电针+乌头碱组大鼠经左股静脉注射乌头碱溶液（10 μg/kg）。检测各组大鼠血流动力学指标左心室收缩压（LVSP）、左心室内压最大上升/下降变化速率（±dp/dtmax）；观察大鼠心律失常类型并进行评分；多重荧光Western blot法检测大鼠左心室心肌组织SERCA2a和PLB蛋白表达。结果：与对照组比较，模型组造模后、干预后各时点LVSP、±dp/dtmax 均降低（P<0.01）；与模型组比较，乌头碱组、电针组干预后1 min ±dp/dtmax升高（P<0.01，P<0.05），乌头碱组干预后10~60 min与电针组干预后20~60 min +dp/dtmax升高（P<0.01，P<0.05），电针组干预后1 min LVSP亦升高（P<0.01），电针+乌头碱组干预后1~60 min LVSP、±dp/dtmax 均升高（P<0.01，P<0.05）。与乌头碱组比较，电针组干预后1 min LVSP和+dp/dtmax升高（P<0.01，P<0.05），电针+乌头碱组干预后1 min LVSP、±dp/dtmax与干预后20~60 min +dp/dtmax均升高（P<0.01，P<0.05）。与电针组比较，电针+乌头碱组干预后10~60 min +dp/dtmax升高（P<0.01）。与模型组比较，乌头碱组干预后心律失常评分升高（P<0.01）；与乌头碱组比较，电针+乌头碱组与电针组心律失常评分降低（P<0.01）。与对照组比较，模型组左心室心肌组织SERCA2a蛋白表达降低（P<0.01），PLB蛋白表达增加（P<0.01）；与模型组比较，电针组和电针+乌头碱组SERCA2a蛋白表达增加（P<0.05，P<0.01），各干预组PLB蛋白表达降低（P<0.01，P<0.05）；与电针组、乌头碱组比较，电针+乌头碱组SERCA2a蛋白表达增加、PLB蛋白表达降低（P<0.05，P<0.01）。结论：电针干预可能是通过上调心肌组织SERCA2a蛋白表达、下调PLB蛋白表达实现对乌头碱改善心力衰竭的增效/减毒作用。.
Keywords: aconitine; calcium pump; combination of acupuncture and medication; electroacupuncture; heart failure; synergistic and attenuated effect.