葡萄籽原花青素对缺血性心力衰竭大鼠心肌细胞凋亡及MAPK/ERK1/2通路的影响

目的:观察葡萄籽原花青素(GSPE)对缺血性心力衰竭(IHF)大鼠心肌功能的作用,初步探讨其作用机制。方法:构建缺血性心力衰竭模型大鼠,随机分为5组:IHF组,卡普托利组,低、中、高剂量GSPE组,每组15只,另设对照组15只。低、中、高剂量GSPE组大鼠分别采用50、100、200 mg/kg GSPE灌胃给药,卡普托利组采用300 mg/kg卡普托利灌胃给药,对照组和模型组采用等体积生理盐水灌胃,各组大鼠均每天定时灌胃给药1次,连续4周。多普勒彩色超声心动图进行心脏血流动力学测定;苏木素(H)-伊红(E)染色观察心肌组织病变程度;TUNEL法检测心肌细胞凋亡情况;蛋白免疫印迹(WB)检测心肌组织半胱氨酸蛋白酶cleaved-caspase3、cleaved-caspase9及C-Jun N-末端激酶(JNK)、有丝分裂原活化蛋白激酶1/2(ERK1/2)、细胞外信号调节激酶(MAPK)蛋白表达情况。结果:与对照组比较,IHF大鼠HR、LVEDP、凋亡细胞比例均显著升高(P<0.05),MAP、LVSP显著降低(P<0.05);心肌组织可见明显病理损伤,cleaved-caspase3、cleaved-caspase9蛋白及ERK1/2、MAPK、JNK蛋白磷酸化水平明显增高(P<0.05)。与IHF组比较,低、中、高剂量GSPE组和卡普托利组大鼠HR、LVEDP、凋亡细胞比例明显降低,MAP、LVSP显著升高(P<0.05);心肌组织病理损伤改善,cleaved-caspase3、cleaved-caspase9蛋白及ERK1/2、MAPK、JNK蛋白磷酸化水平明显降低(P<0.05)。结论:葡萄籽原花青素可能通过调节MAPK通路抑制心肌细胞凋亡,改善缺血性心力衰竭大鼠心脏功能。

Effects of grape seed proanthocyanidin extract on cardiomyocyte apoptosis and MAPK/ERK1/2 pathway in rats with ischemic heart failure

Objective: To observe the effect of grape seed proanthocyanidin extract(GSPE) on myocardial function of rats with ischemic heart failure(IHF),and to explore its mechanism. Methods: The rat model of ischemic heart failure was constructed and divided into 5 groups randomly:IHF group,Captopril group,low,middle and high dose GSPE groups,with 15 rats in each group,and another 15 rats were set as control group.Rats in low,middle and high dose GSPE groups were administrated with 50 mg/kg,100 mg/kg, 200 mg/kg GSPE respectively.Captopril group was given captopril 300 mg/ml by gastric administration,while control group and model group were given equal volume normal saline by gastric administration.Rats in each group were given intragastric administration once a day for 4 weeks.Doppler echocardiography was used to determine the hemodynamics of the heart.Hematoxylin(H)-eosin(E)staining was used to observe the degree of myocardial tissue lesion.TUNEL method was used to detect cardiomyocyte apoptosis.The expressions of cleaved-caspase3,cleaved-caspase9,C-Jun N-terminal kinase(JNK),mitogen activated protein kinase 1/2(ERK1/2),extracellular signal regulated kinase(MAPK) in myocardial tissues were detected by protein immunoblotting(WB). Results: Compared with the control group,HR,LVEDP and the proportion of apoptotic cells in IHF rats increased significantly( P <0.05),and MAP,LVSP decreased significantly( P <0.05);there was obvious pathological injury in the myocardium;the expressions of cleaved-caspase3 and cleaved-caspase9 were significantly increased( P <0.05);the phosphorylation levels of ERK1/2,MAPK and JNK proteins were significantly increased( P <0.05).Compared with the IHF group,HR,LVEDP and the proportion of apoptotic cells in the low,middle and high dose GSPE groups and the Captopril group decreased significantly,and MAP,LVSP increased significantly( P <0.05);the pathological injury of myocardial tissue was improved;the expressions of cleaved-caspase3 and cleaved-caspase9 were significantly decreased( P <0.05);the phosphorylation levels of ERK1/2,MAPK and JNK proteins were significantly decreased( P <0.05). Conclusion: Grape seed proanthocyanidin extracts may inhibit cardiomyocyte apoptosis by regulating the MAPK pathway,and then improve the cardiac function in rats with ischemic heart failure.