Puerarin Attenuates Anoxia/Reoxygenation Injury Through Enhancing Bcl-2 Associated Athanogene 3 Expression, a Modulator of Apoptosis and Autophagy.
Puerarin Attenuates Anoxia/Reoxygenation Injury Through Enhancing Bcl-2 Associated Athanogene 3 Expression, a Modulator of Apoptosis and Autophagy.
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DOI：
10.12659/msm.897379
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发表时间：
2016-03-24
期刊：
影响因子：
--
通讯作者：
Zhang Y
中科院分区：
文献类型：
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作者：
Ma Y;Gai Y;Yan J;Li J;Zhang Y
葛根素对缺血再灌注损伤具有保护作用，但其机制尚未完全阐明。本研究旨在探讨葛根素对体外培养乳鼠原代心肌细胞缺氧/复氧损伤（A/RI）中Bcl-2相关凋亡基因3（BAG 3）表达的影响及其在A/RI中的作用。使用qRT-PCR和蛋白质印迹分析定量有或没有葛根素预处理的心肌细胞中的BAG 3表达。通过测定乳酸脱氢酶（LDH）、磷酸肌酸激酶（CPK）活性、丙二醛（MDA）、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）含量，观察BAG 3对A/RI的影响。进一步研究BAG 3对A/RI后心肌细胞自噬和凋亡的影响。葛根素显著促进A/RI后大鼠原代心肌细胞BAG 3的表达。在CPK、LDH、MDA、SOD、GSH-Px、ROS生成和细胞活力方面，加强的BAG 3表达在减弱A/RI方面表现出与葛根素预处理相似的效果。在A/RI后，BAG 3过表达显著刺激心肌细胞的自噬，这在细胞活力和凋亡方面对A/RI具有保护作用。自噬抑制部分取消了BAG 3的保护作用。葛根素可直接增加A/RI后心肌细胞BAG 3的转录和翻译。BAG 3表达的增加至少通过增强自噬和减少凋亡对A/RI具有保护作用，这是葛根素在ARI中的新的保护机制。
Puerarin has protective effects on ischemia-reperfusion injury, but the underlying mechanisms are not fully revealed. This study explored the effect of puerarin on the expression of Bcl-2 associated athanogene 3 (BAG3) in an in vitro model of anoxia/reoxygenation injury (A/RI) in neonate rat primary cardiomyocytes and the functions of BAG3 in A/RI. BAG3 expression in cardiomyocytes with or without puerarin pre-treatment was quantified using qRT-PCR and Western blot analysis. The effects of BAG3 on A/RI were studied by measuring the activity of lactate dehydrogenase (LDH) and creatine phosphate kinase (CPK), the concentration of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px). The effects of BAG3 on autophagy and apoptosis of the cardiomyocytes after A/RI were further studied. Puerarin significantly promoted BAG3 expression in the rat primary cardiomyocytes after A/RI. Enforced BAG3 expression presented similar effects as puerarin pre-treatment in attenuating A/RI in terms of CPK, LDH, MDA, SOD, GSH-Px, ROS generation, and cell viability. BAG3 overexpression significantly stimulated autophagy in cardiomyocytes after A/RI, which presented protective effects on A/RI in terms of cell viability and apoptosis. Autophagy inhibition partly abrogated the protective effects of BAG3. Puerarin can directly increase BAG3 transcription and translation in cardiomyocytes after A/RI. The elevated BAG3 expression presents protective effects on A/RI at least through enhancing autophagy and reducing apoptosis, which is a novel protective mechanism of puerarin in ARI.