Protective effects of upregulated HO-1 gene against the apoptosis of human retinal pigment epithelial cells in vitro

Abstract

Aim: To investigate the protective effect of heme oxygenase-1 (HO-1) against H₂O₂-induced apoptosis in human ARPE-19 cells.

Methods: The lentiviral vector expressing HO-1 was prepared and transfected into apoptotic ARPE-19 cells induced by H₂O₂. Functional experiments including cell counting kit-8 (CCK-8) assay, flow cytometry (FCM) and mitochondrial membrane potential assay were conducted.

Results: The ultrastructure of ARPE-19 cells was observed using transmission electron microscope (TEM). It was found that exogenous HO-1 significantly ameliorated H₂O₂-induced loss of cell viability, apoptosis and intracellular levels of reactive oxygen species (ROS) in ARPE-19 cells. The overexpression of HO-1 facilitated the transfer of nuclear factor erythroid-2-related factor 2 (Nrf2) from cytoplasm to nucleus, which in turn upregualted expressions HO-1 and B-cell lymphoma-2 (Bcl-2). Furthermore, HO-1 upregulation further inhibited H₂O₂-induced release of cysteinyl aspartate specific proteinase-3 (caspase-3).

Conclusion: Exogenous HO-1 protect ARPE-19 cells against H₂O₂-induced oxidative stress by regulating the expressions of Nrf2, HO-1, Bcl-2, and caspase-3.

Keywords: ARPE-19 cell; heme oxygenase-1; mechanism; retinitis pigmentosa.