Ferroptosis and necroptosis may be involved in the formation and progression of hydrofluoric acid burn wounds: Results from an RNA-Seq analysis.

Abstract

Hydrofluoric acid (HF) burns have potentially serious consequences. The molecular mechanism of wound development is still unclear. This study aims to preliminarily explore the programmed cell death mode that may be involved in hydrofluoric acid burns by using transcriptome sequencing technology and to provide a theoretical basis for a new treatment approach for hydrofluoric acid burns. The rat model of hydrofluoric acid burn skin was constructed, and the differentially expressed genes after HF burn were screened by transcriptome sequencing technology. HE staining, TUNEL staining, immunohistochemistry, biochemical detection, and qRT-PCR were used to preliminarily verify the mode of cell death involved in hydrofluoric acid burn wounds. The sequencing results suggest that the differential genes after HF burn were enriched in ferroptosis, apoptosis, and necroptosis pathways in cell growth and death aspects. HE staining confirmed HF burn wounds were progressively aggravated. The positive cells of TUNEL staining in the wound gradually increased. Compared with the normal group, the content of MDA in serum and skin tissue increased and the content of GSH decreased at 4, 8, 12, 24, and 48 hours after HF burn (P < 0.05). The level of serum Fe^2 + in the HF burn group was higher than that in the normal group at 4 h, 8 h, and 12 h postburn (P < 0.05). The level of serum Fe²⁺ at 24 h and 48 h postburn was higher than that of the normal group, but the difference was not statistically significant. The content of Fe²⁺ in skin tissue increased and reached its peak at 12 h (P < 0.05). The serum calcium level decreased to its lowest level at 24 hours postburn (P < 0.05). Immunohistochemistry showed that the expressions of GPX4, FTH1, and Bcl-2 proteins in hydrofluoric acid burn wounds were down-regulated, while the expression of HO-1, Bax, RIPK1, and MLKL was increased (P < 0.05). RIPK3 expression was not significantly different. qRT-PCR showed that the expression of HO-1, FTH1, SLC39A14, SLC39A8, CYBB, ACSL4, Bax, RIPK1, MLKL, IL-1β, and IL-6 increased, while the expression of ACSL1, ACSL6, GPX4, and Bcl-2 decreased after hydrofluoric acid burn compared with the normal group (P < 0.05). The RIPK3 gene expression did not change significantly. Ferroptosis and necroptosis are involved in the formation and progression of HF burn wounds. Early blocking of ferroptosis may be a potential therapeutic for blocking the progress of hydrofluoric acid burn wounds. Necroptosis involvment in the occurrence and development of hydrofluoric acid burn wounds may be a non-classical pathway.