Variation of Ferroptosis-Related Markers in HaCaT Cell Photoaging Models Induced by UVB

Clin Cosmet Investig Dermatol. 2023 Nov 1:16:3147-3155. doi: 10.2147/CCID.S433071. eCollection 2023.

Abstract

Objective: To investigate the variation of ferroptosis-related markers in HaCaT cell photoaging models induced by ultraviolet-B (UVB).

Methods: UVB-treated HaCaT cells served as the model (UVB group) for cellular photoaging, whereas untreated HaCaT cells served as the control group. HaCaT cells were exposed to UVB and the ferroptosis inhibitor Ferrostatin-1 (Fer-1) as part of the UVB+Fer-1 group, and co-cultured with the ferroptosis inducer Erastin as part of the UVB+Erastin group. Reactive oxygen species (ROS) detection kit and senescence-related β galactosidase (SA-β-gal) staining were used to evaluate the senescence of HaCaT cells. Lipid reactive oxygen species were detected by C11 BODIPY581/591 probe and mitochondrial morphology was observed by transmission electron microscopy. The mRNA expressions of glutathione peroxidase 4 (GPX4) and ferroptosis-suppressor-protein 1 (FSP1) were detected by real-time reverse transcription-PCR (RT-RCP), and the level of GPX4 protein was measured by immunofluorescence assay.

Results: The UVB group had considerably greater levels of ROS, SA-β-gal, and lipid reactive oxygen species than the control group. The UVB group's mitochondrial volume was reduced, the membrane density increased, and the mitochondrial crest decreased or even disappeared. GPX4 and FSP1 expression levels were similarly found to be lower in the UVB group. Furthermore, the positive rate of SA-β-gal and lipid reactive oxygen species in the UVB+Fer-1 group was much lower than in the UVB group, but it was reverse in the UVB+Erastin group. This study showed that induced ferroptosis can aggravate aging, and vice versa.

Conclusion: According to the findings, ferroptosis may be linked to UVB-induced skin photoaging, which could be attenuated by inhibition of ferroptosis.

Keywords: ferroptosis; oxidative stress; skin aging.

Grants and funding

National Natural Science Foundation of China (81674039, 81873347); Key Projects of Scientific Research Fund of Traditional Chinese Medicine of Hubei Health Commission (ZY2021Z011); Training Project Funding Plan of Young-aged Talent of Health System in Hubei Province (2020–2023); Key Projects of Chongqing Municipal Health Commission and Science and Technology Bureau Joint Medical Research (2022ZDXM037); Key Projects of Traditional Chinese Medicine of Wuhan Municipal Health Commission (WZ22A03).