PMID: 15238173Jul 9, 2004Paper

Role of bcl-2 and bax expression in apoptosis of hypoxic pulmonary artery smooth muscle cells of rats induced by Na+/H+ exchanger isoform-1 inhibition in vitro

Zhongguo wei zhong bing ji jiu yi xue = Chinese critical care medicine = Zhongguo weizhongbing jijiuyixue
Junyu LuShu-Ping Li

Abstract

To investigate the role of bcl-2 and bax expression in apoptosis of hypoxic pulmonary artery smooth muscle cells (PASMCs) of rats induced by Na+/H+ exchanger isoform-1 (NHE-1) inhibition in vitro. Cultured rat PASMCs transfected with NHE-1 ribozyme gene (PRZ cell), cultured rat PASMCs transfected only with pLXSN vector (PX cell) and non-transfected rat PASMCs (PA cell) were obtained in our previous studies. The cells were cultured under low oxygen tension (<1% O2) for 0, 2, 6, 12, 24 and 48 hours respectively. Then cell apoptosis was examined with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). Intracellular Ca2+([Ca2+]i) of cells was measured using fluorescence dye Fura-2/AM. The changes in mRNA expression of bcl-2 and bax in cells were assessed by semi quantitative reverse transcription-polymerase chain reaction (sqRT-PCR). The expression of Bcl-2 protein and Bax protein of cells was detected immunohistochemically. In contrast to normal PASMCs and cells transfected with pLXSN, the apoptosis rates in recombinant vectors transfected cells increased significantly and continuously. [Ca2+]i values were also increased in these cells, but the increase extent was remarkably lower than in the normal PASM...Continue Reading

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