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.

Fisetin Attenuated Oxidative Stress-Induced Cellular Damage in ARPE-19 Human Retinal Pigment Epithelial Cells Through Nrf2-Mediated Activation of Heme Oxygenase-1

“…This is in good agreement with the results of previous studies ( Zhao et al, 2019 ; Rodius et al, 2020 ; Molagoda et al, 2021 ; Ding et al, 2022 ) and suggests that fisetin may act as an oxidative stress blocker. Several previous studies have proven that oxidative stress-induced apoptosis by H 2 O 2 treatment in ARPE-19 cells is directly correlated with mitochondrial impairment ( Zhao et al, 2020 ; Hong et al, 2021b ; Kim et al, 2021b ). Alteration of the outer mitochondrial membrane permeability by oxidative damage is linked to MMP dissipation.…”

“…For example, it has been reported that negative regulation of Nrf2 is implicated in the pathogenesis of maculopathy ( Datta et al, 2017 ; Bellezza, 2018 ; Hyttinen et al, 2019 ), and mice deficient in Nrf2 have been shown to exhibit retinal alterations similar to AMD ( Zhao et al, 2014 ; Rowan et al, 2020 ). On the other hand, Nrf2-mediated activation of HO-1 was effective in ameliorating oxidative damage in human RPE cells ( Lambros and Plafker, 2016 ; Hyttinen et al, 2019 ; Hong et al, 2021b ). In the meantime, many previous results have demonstrated that activation of HO-1 by some phytochemicals with antioxidant activity plays an important role in the resistance of ARPE-19 cells to oxidative stress ( Zhao et al, 2020 ; Chen et al, 2021 ; Zhang et al, 2022 ).…”

“…In this context, protecting RPE cells from oxidative stress could be an effective strategy to prevent or attenuate retina-mediated ophthalmic diseases. Heme oxygenase-1 (HO-1) is a responsive enzyme that catabolizes free heme into carbon monoxide, and the degradation of HO-1 into heme consumes 3 molecules of oxygen, indirectly triggering an inhibitory effect on oxidative damage ( Lenzi et al, 2005 ; Hong et al, 2021a ). Accumulated evidences suggested that HO-1 can be activated by various forms of stimuli to protect cells including RPE ( Zhou et al, 2020a ; Hong et al, 2021a ).…”

“…Heme oxygenase-1 (HO-1) is a responsive enzyme that catabolizes free heme into carbon monoxide, and the degradation of HO-1 into heme consumes 3 molecules of oxygen, indirectly triggering an inhibitory effect on oxidative damage ( Lenzi et al, 2005 ; Hong et al, 2021a ). Accumulated evidences suggested that HO-1 can be activated by various forms of stimuli to protect cells including RPE ( Zhou et al, 2020a ; Hong et al, 2021a ). There has been reported that activation of nuclear factor erythroid-2-related factor 2 (Nrf2)/HO-1 attenuated apoptosis in APRE-19 human RPE cell line ( Zhou et al, 2020a ; Hong et al, 2021a ).…”

“…Accumulated evidences suggested that HO-1 can be activated by various forms of stimuli to protect cells including RPE ( Zhou et al, 2020a ; Hong et al, 2021a ). There has been reported that activation of nuclear factor erythroid-2-related factor 2 (Nrf2)/HO-1 attenuated apoptosis in APRE-19 human RPE cell line ( Zhou et al, 2020a ; Hong et al, 2021a ).…”