title: Small molecule antidepressant amitriptyline protects hypoxia/reoxygenation-induced cardiomyocyte apoptosis through TrkA signaling pathway.
abstract: 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 2h 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), phosphor-TrkA (p-TrkA), protein kinase B (Akt) and phosphor-Akt (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.
authors.affiliation: Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
authors.forename: Yuxiang
authors.initials: Y
authors.lastname: Dai
authors.affiliation: Department of Cardiology, The first People's Hospital of Wenling, Wenling, China.
authors.forename: Shen
authors.initials: S
authors.lastname: Wang
authors.affiliation: Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
authors.forename: Chenguang
authors.initials: C
authors.lastname: Li
authors.affiliation: Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
authors.forename: Shufu
authors.initials: S
authors.lastname: Chang
authors.affiliation: Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Lu
authors.affiliation: Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
authors.forename: Zheyong
authors.initials: Z
authors.lastname: Huang
authors.affiliation: Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
authors.forename: Hongbo
authors.initials: H
authors.lastname: Yang
authors.affiliation: Department of Gerontology, Zhongshan Hospital, Fudan University, Shanghai, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Shen
authors.affiliation: Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
authors.forename: Zhangwei
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
authors.forename: Juying
authors.initials: J
authors.lastname: Qian
authors.affiliation: Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai 200032, China. Electronic address: zhangg5@aol.com.
authors.forename: Junbo
authors.initials: J
authors.lastname: Ge
mesh_terms: D000639:Amitriptyline / Q000494:pharmacology*; D000818:Animals; D000928:Antidepressive Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D015687:Cell Hypoxia / Q000187:drug effects; D019610:Cytoprotection / Q000187:drug effects; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism*; D020917:Receptor, trkA / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*
chemical_list: D000928:Antidepressive Agents; D000639:Amitriptyline; D020917:Receptor, trkA; D010100:Oxygen
keywords: Amitriptyline; Apoptosis; Cardiomyocyte; Hypoxia; Reoxygenation; TrkA