title: Na(+)/Ca(2+) exchanger 1 (NCX-1) mediates the anti-apoptotic effect of Akt1 in neonatal rat cardiomyocytes during ischemia/reperfusion.
abstract: The purpose of this study was to investigate the anti-apoptotic role of Akt1 gene in neonatal rat cardiomyocytes and the relationship with Na(+)/Ca(2+) exchanger 1 (NCX1) during ischemia/reperfusion (IR). The cultured original rat cardiomyocytes were randomly divided into five groups: normal control group (C group), hypoxia/reoxygenation group (HR group), the control vector pLVX-EGFP-3FLAG group (CV group), the gene pLVX-EGFP-3FLAG-Akt1 transfection group (A group), and Akt1 inhibitor LY294002 group (LY group). Cardiomyocyte vitality was determined using MTT, and the apoptosis was determined by TUNEL to verify the anti-apoptotic role of Akt1. The mRNA levels of Akt1 and NCX1 were determined by RT-PCR, the protein expression of Akt1, p-Akt1, NCX1 and the apoptotic proteins of mitochondrial pathway cytochrome C (Cyto C) and caspase-9 were measured by Western blot. As a result, transfected Akt1 (A group) showed increased myocardial cell viability and reduced apoptosis, with increase in Akt1 expression and decrease in NCX1 expression. The levels of apoptotic proteins Cyto C and caspase-9 also declined. This study demonstrated that lentivirus-mediated transfection of Akt1 played an anti-apoptotic role during IR of rat cardiomyocytes, via inhibition of NCX1 and other mitochondrial proteins.
authors.affiliation: Institute of Cardiovascular Disease Research, Xuzhou Medical University Xuzhou 221002, Jiangsu, China.
authors.forename: Manman
authors.initials: M
authors.lastname: Huang
authors.affiliation: Affiliated Hospital of Xuzhou Medical University Xuzhou 221002, Jiangsu, China.
authors.forename: Defeng
authors.initials: D
authors.lastname: Pan
authors.affiliation: Institute of Cardiovascular Disease Research, Xuzhou Medical University Xuzhou 221002, Jiangsu, China.
authors.forename: Yinping
authors.initials: Y
authors.lastname: Du
authors.affiliation: Affiliated Hospital of Xuzhou Medical University Xuzhou 221002, Jiangsu, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Zhu
authors.affiliation: Institute of Cardiovascular Disease Research, Xuzhou Medical University Xuzhou 221002, Jiangsu, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Affiliated Hospital of Xuzhou Medical University Xuzhou 221002, Jiangsu, China.
authors.forename: Tongda
authors.initials: T
authors.lastname: Xu
authors.affiliation: Affiliated Hospital of Xuzhou Medical University Xuzhou 221002, Jiangsu, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Luo
authors.affiliation: Institute of Cardiovascular Disease Research, Xuzhou Medical UniversityXuzhou 221002, Jiangsu, China; Affiliated Hospital of Xuzhou Medical UniversityXuzhou 221002, Jiangsu, China.
authors.forename: Dongye
authors.initials: D
authors.lastname: Li
keywords: Akt1; NCX1; apoptosis; cardiomyocytes; ischemia/reperfusion