Small molecule antidepressant amitriptyline protects hypoxia/ reoxygenation-induced cardiomyocyte apoptosis through TrkA signaling pathway
Small molecule antidepressant amitriptyline protects hypoxia/ reoxygenation-induced cardiomyocyte apoptosis through TrkA signaling pathway
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DOI：
10.1016/j.ejphar.2017.01.029
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发表时间：
2017-03-05
影响因子：
5
通讯作者：
Ge, Junbo
中科院分区：
文献类型：
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作者：
Dai, Yuxiang;Wang, Shen;Ge, Junbo
缺氧/复氧（Hypoxia/reoxygenation，H/R）诱导的心肌细胞凋亡是导致心血管疾病的重要因素。在这项研究中，我们研究了小分子抗抑郁药阿米替林（AMP）对H/R诱导的新生小鼠心肌细胞凋亡的保护作用。用H/R处理C57 BL/6 J小鼠心肌细胞。在H/R条件化前2 h向培养物中加入不同浓度的AMP。TUNEL法检测心肌细胞凋亡。蛋白质印迹法检测AMP诱导的下游信号通路蛋白，包括原肌球蛋白受体激酶A受体（TrkA）、磷酸化TrkA（p-TrkA）、蛋白激酶B（Akt）和磷酸化Alit（p-Akt）。然后用K252 a阻断TrkA磷酸化，以研究TrkA是否在功能上参与AMP对H/R损伤的心肌细胞的保护。我们发现，H/R条件下诱导显著的心肌细胞死亡和凋亡，而AMP预处理相当大地挽救心肌细胞死亡和凋亡。Western blot分析表明AMP通过磷酸化TrkA/Akt蛋白激活TrkA信号通路。K252 a抑制TrkA/Akt信号通路的磷酸化，从而消除AMP对H/R诱导的心肌细胞凋亡的保护作用。因此，我们的研究表明，AMP通过激活TrkA/Akt信号通路，对H/R诱导的心肌细胞凋亡起保护作用。
Hypoxia/reoxygenation (H/R) induced cardiomyocytes apoptosis is a major factor leading to cardiovascular diseases. In this study, we investigated the protective effect of small molecule antidepressant amitriptyline (AMP) in regulating H/R-induced apoptosis in neonatal mouse cardiomyocyte in culture. Cardiomyocytes of C57BL/6J mice were treated with H/R condition in vitro. Various concentration of AMP was added into culture 2 h prior to H/R conditioning. Cardiomyocyte apoptosis was evaluated by TUNEL assay. AMP induced downstream signaling pathway proteins, including tropomyosin receptor kinase A receptor (TrkA), phosphorTrkA (p-TrkA), protein kinase B (Akt) and phosphor -Alit (p-Akt) were probed -by western blot. TrkA phosphorylation was then blocked by K252a to investigate whether TrkA was functionally involved in the protection of AMP in H/R-injured cardiomyocyte. We found that H/R condition induced significant cardiomyocyte death and apoptosis, whereas AMP pretreatment considerably rescued cardiomyocyte death and apoptosis. Western blot analysis showed AMP activated TrkA signaling pathway through the phosphorylation of TrkA/Akt proteins. We also found that application of K252a inhibited the phosphorylation of TrkA/Akt signaling pathway, and subsequently abolished the protective effect of AMP in H/R-induced apoptosis in cardiomyocyte. Thus, our study revealed that AMP, through the activation of TrkA/Akt signaling pathway, plays a protective role in regulating H/R-induced apoptosis in cardiomyocyte.