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目的:明确苓桂养心汤对扩张型心肌病(DCM)大鼠心功能、心肌的保护作用;观察苓桂养心汤对DCM大鼠损伤心肌肌浆网(SR)钙调控相关蛋白的影响,探讨其可能的作用机制。方法:以60只SD大鼠腹腔注射阿霉素1.0 mg/kg/次,每周2次,连续6周,总量12 mg·kg-1建立DCM模型,另设正常组(10只),观察2周,8周后将存活DCM模型大鼠随机分为培哚普利组、苓桂养心汤组、模型组,连续灌胃给药8周。治疗后对上述各组大鼠行心脏超声检查;采血测定大鼠血清B型尿钠肽(BNP)水平及肌钙蛋白-Ⅰ(cTnI);取大鼠左心室心肌进行苏木素-伊红(HE)染色和透射电子显微镜检测大鼠心肌病理学形态。采用逆转录聚合酶链式反应(RT-PCR)和蛋白质免疫印迹法(Western blot)分别测定大鼠心肌肌浆网钙泵(SERCA2a),钙释放通道(RyR2),受磷蛋白(PLB),肌集钙蛋白(Calsequestrin),L-型钙离子通道(CAV1.3)的mRNA及蛋白表达水平。结果:与模型组比较,苓桂养心汤组可以改善DCM模型大鼠的心腔大小、心功能、心肌损伤及心肌病理学形态。与模型组比较,苓桂养心汤组SERCA2a,RyR2,CAV1.3,Calsequestrin mRNA表达升高;PLB mRNA表达下降。与模型组比较,苓桂养心汤组SERCA2a及Calsequestrin蛋白表达升高;PLB蛋白表达下降。结论:苓桂养心汤能缩小DCM大鼠左心室内径,提高左心室射血分数和缩短率,降低血清BNP和cTnI水平,保护DCM大鼠损伤心肌,改善心功能。苓桂养心汤可部分调控DCM大鼠心肌肌浆网钙泵,兰尼碱受体,受磷蛋白,肌集钙蛋白,L-型钙离子通道的mRNA基因和蛋白表达。苓桂养心汤治疗DCM大鼠的作用机制,可能与通过调节DCM大鼠心肌肌浆网钙调控相关蛋白的基因及蛋白表达,改善心肌细胞钙转运及心脏舒缩功能有关。
Objective: To clarify the protective effects of Linggui Yangxin Decoction on cardiac function and myocardium in DCM rats, and to observe the effect of Linggui Yangxin Decoction on the Ca2 + -regulated proteins of cardiac muscle sarcoplasmic reticulum (SR) in DCM rats Its possible mechanism of action. Methods: Sixty Sprague-Dawley rats were injected intraperitoneally with doxorubicin 1.0 mg / kg twice a week for 6 weeks for a total of 12 mg · kg-1 to establish a DCM model. A normal group (n = 10) After 2 weeks and 8 weeks, the surviving DCM rats were randomly divided into perindopril group, Linggui Yangxin Decoction group and model group, and were given continuous gavage for 8 weeks. After treatment, the rats in each group were subjected to echocardiography. Blood samples were taken to determine the level of serum BNP and cTnI. Left ventricular myocardium was taken for hematoxylin-eosin (HE) ) Staining and transmission electron microscopy. Reverse transcription polymerase chain reaction (RT-PCR) and Western blot were used to detect the expression of SERCA2a, RyR2, PLB, Calsequestrin, L-type calcium channel (CAV1.3) mRNA and protein expression levels. Results: Compared with the model group, Ling-Gui Yang-Xin Decoction group can improve the cardiac cavity size, cardiac function, myocardial injury and pathological morphology in DCM rats. Compared with the model group, the expressions of SERCA2a, RyR2, CAV1.3 and Calsequestrin mRNA in Ling-ling Yang-xin Decoction group were increased, while the expression of PLB mRNA was decreased. Compared with the model group, the expression of SERCA2a and Calsequestrin in Linggui Yangxin Decoction group increased and the expression of PLB protein decreased. Conclusion: Lingui Yangxin Decoction can reduce left ventricular diameter of DCM rats, increase left ventricular ejection fraction and shortening rate, lower serum BNP and cTnI levels, protect myocardium of DCM rats and improve cardiac function. Linggui Yangxin Decoction can partially regulate the mRNA and protein expression of sarcoplasmic reticulum calcium pump, ryanodine receptor, phospholamban, calcitonin and L-type calcium channel in DCM rats. The mechanism of Linggui Yangxin Decoction in treating DCM rats may be related to the regulation of calcium transport and cardiac systolic and diastolic function in cardiomyocytes by regulating the gene and protein expression of calcium regulation related proteins in cardiac sarcoplasmic reticulum of DCM rats.