Chen Chuanjun, He Huan, Luo Yong, Zhou Min, Yin Dong, He Ming
*Jiangxi Provincial Key Laboratory of Basic Pharmacology, Nanchang University School of Pharmaceutical Science, Nanchang, China; and †Jiangxi Provincial Key Laboratory of Molecular Medicine at the Second Affiliated Hospital, Nanchang University, Nanchang, China.
J Cardiovasc Pharmacol. 2016 Feb;67(2):152-63. doi: 10.1097/FJC.0000000000000331.
Apigenin is a type of flavonoids, which has been demonstrated to protect myocardium against ischemia/reperfusion (I/R) injury. However, the mechanism is still unclear. We hypothesized that the mechanism of cardioprotective action of apigenin on the I/R-induced injury might be caused via B-cell lymphoma (Bcl) signaling pathway. In this study, an in vitro I/R model was replicated on Langendorff-perfused heart and H9c2 cardiomyocytes by anoxia/reoxygenation (A/R) treatment. The recovery of cardiac contractile function, infarct size, lactate dehydrogenase (LDH) and creatine kinase (CK) in the perfusate, the expression and activity of Bcl-2 and caspase-3, and cardiomyocyte apoptosis were measured in the Langendorff heart undergoing A/R injury. In addition, the cell viability, LDH release, intracellular reactive oxygen species (ROS), mitochondrial membrane potential (Δψm), expression of cytochrome c in the cytosol, and cell apoptosis were examined in the culture of H9c2 cardiomyocytes after the A/R. The results showed that apigenin significantly improved rat heart contractile function, reduced LDH release, infarct size and apoptotic rate, upregulated the expression of Bcl-2 and caspase-3, and downregulated the expression of cleaved caspase-3 after the A/R. Moreover, apigenin increased the cell viability and decreased the release of LDH, production of reactive oxygen species, release of mitochondrial cytochrome c into the cytosol, and cell apoptosis in the culture of H9c2 cardiomyocytes after the A/R. In addition, inhibition of Bcl-2 activity by ABT-737 markedly attenuated the protective effect of apigenin on the A/R-induced myocardium injury. Taken together, we firstly demonstrated that the effect of apigenin against A/R injury in cardiomyocytes involves Bcl-2 signal pathway and at least partly depends on its effect of upregulating the expression of Bcl-2.
芹菜素是一种黄酮类化合物，已被证明可保护心肌免受缺血/再灌注（I/R）损伤。然而，其机制仍不清楚。我们推测芹菜素对I/R诱导损伤的心脏保护作用机制可能是通过B细胞淋巴瘤（Bcl）信号通路介导的。在本研究中，通过缺氧/复氧（A/R）处理在Langendorff灌注心脏和H9c2心肌细胞上复制了体外I/R模型。测量了经历A/R损伤的Langendorff心脏的心脏收缩功能恢复、梗死面积、灌注液中乳酸脱氢酶（LDH）和肌酸激酶（CK）、Bcl-2和半胱天冬酶-3的表达及活性以及心肌细胞凋亡情况。此外，检测了A/R后H9c2心肌细胞培养物中的细胞活力、LDH释放、细胞内活性氧（ROS）、线粒体膜电位（Δψm）、细胞溶质中细胞色素c的表达以及细胞凋亡。结果显示，芹菜素显著改善了大鼠心脏收缩功能，降低了LDH释放、梗死面积和凋亡率，上调了A/R后Bcl-2和半胱天冬酶-3的表达，并下调了裂解的半胱天冬酶-3的表达。此外，芹菜素增加了细胞活力，降低了A/R后H9c2心肌细胞培养物中的LDH释放、活性氧产生、线粒体细胞色素c释放到细胞溶质中以及细胞凋亡。此外，ABT-737抑制Bcl-2活性显著减弱了芹菜素对A/R诱导的心肌损伤的保护作用。综上所述，我们首次证明芹菜素对心肌细胞A/R损伤的作用涉及Bcl-2信号通路，并且至少部分取决于其上调Bcl-2表达的作用。
