Liguo Yang1, Shigang Song2, Hang Lv1. 1. Second Ward of Cardiovascular Surgery, Second Affiliated Hospital of Harbin Medical University Harbin 157000, China. 2. Department of Radiation Oncology, Second Affiliated Hospital of Harbin Medical University Harbin 157000, China.
Abstract
BACKGROUND: Cardiomyocytes apoptosis under hypoxia condition contributes significantly to various cardiovascular diseases. In this study, we investigated the role of microRNA-322 (miR-322) in regulating hypoxia-induced apoptosis in neonatal murine cardiomyocytes in vitro. METHOD: Cardiomyocytes of C57BL/6J mice were treated with hypoxia condition in vitro. Cardiomyocyte apoptosis was measured by TUNEL assay. Gene expression pattern of miR-322 was measured by qRT-PCR. Stable downregulation of miR-322 in cardiomyocytes were achieved by lentiviral transduction, and the effect of miR-322 downregulation on hypoxia-induced cardiomyocyte apoptosis was investigated. Possible regulation of miR-322 on its downstream target gene, brain derived neurotrophic factor (BDNF) was investigated in cardiomyocytes. BDNF was then genetically silenced by siRNA to evaluate its role in miR-137 mediated cardiomyocyte apoptosis protection under hypoxia condition. RESULTS: Under hypoxia condition, significant apoptosis was induced and miR-322 was significantly upregulated in cardiomyocytes in vitro. Through lentiviral transduction, miR-322 was efficiently knocked down in cardiomyocytes. Downregulation of miR-322 protected hypoxia-induced cardiomyocyte apoptosis. Luciferase assay showed BDNF was the target gene of miR-322. QRT-PCR showed BDNF expression was associated with miR-322 regulation on hypoxia-induced cardiomyocyte apoptosis. Silencing BDNF in cardiomyocyte through siRNA transfection reversed the protective effect of miR-322 downregulation on hypoxia-induced apoptosis. CONCLUSION: Our study revealed that miR-322, in association with BDNF, played important role in regulating hypoxia-induced apoptosis in cardiomyocyte.
BACKGROUND: Cardiomyocytes apoptosis under hypoxia condition contributes significantly to various cardiovascular diseases. In this study, we investigated the role of microRNA-322 (miR-322) in regulating hypoxia-induced apoptosis in neonatal murine cardiomyocytes in vitro. METHOD: Cardiomyocytes of C57BL/6J mice were treated with hypoxia condition in vitro. Cardiomyocyte apoptosis was measured by TUNEL assay. Gene expression pattern of miR-322 was measured by qRT-PCR. Stable downregulation of miR-322 in cardiomyocytes were achieved by lentiviral transduction, and the effect of miR-322 downregulation on hypoxia-induced cardiomyocyte apoptosis was investigated. Possible regulation of miR-322 on its downstream target gene, brain derived neurotrophic factor (BDNF) was investigated in cardiomyocytes. BDNF was then genetically silenced by siRNA to evaluate its role in miR-137 mediated cardiomyocyte apoptosis protection under hypoxia condition. RESULTS: Under hypoxia condition, significant apoptosis was induced and miR-322 was significantly upregulated in cardiomyocytes in vitro. Through lentiviral transduction, miR-322 was efficiently knocked down in cardiomyocytes. Downregulation of miR-322 protected hypoxia-induced cardiomyocyte apoptosis. Luciferase assay showed BDNF was the target gene of miR-322. QRT-PCR showed BDNF expression was associated with miR-322 regulation on hypoxia-induced cardiomyocyte apoptosis. Silencing BDNF in cardiomyocyte through siRNA transfection reversed the protective effect of miR-322 downregulation on hypoxia-induced apoptosis. CONCLUSION: Our study revealed that miR-322, in association with BDNF, played important role in regulating hypoxia-induced apoptosis in cardiomyocyte.
Authors: Goutam Ghosh; Indira V Subramanian; Neeta Adhikari; Xiaoxiao Zhang; Hemant P Joshi; David Basi; Y S Chandrashekhar; Jennifer L Hall; Sabita Roy; Yan Zeng; Sundaram Ramakrishnan Journal: J Clin Invest Date: 2010-10-25 Impact factor: 14.808
Authors: Michele A Hanson; Mohammad Tariq Fareed; Sandra L Argenio; Akochi O Agunwamba; Teresa R Hanson Journal: Prim Care Date: 2013-03 Impact factor: 2.907
Authors: Peter Wohlrab; Lourdes Soto-Gonzales; Thomas Benesch; Max Paul Winter; Irene Marthe Lang; Klaus Markstaller; Verena Tretter; Klaus Ulrich Klein Journal: Front Physiol Date: 2018-12-06 Impact factor: 4.566
Authors: Martin Connolly; Benjamin E Garfield; Alexi Crosby; Nick W Morrell; Stephen J Wort; Paul R Kemp Journal: FEBS Open Bio Date: 2018-01-24 Impact factor: 2.693