Abstract
Abstract and Results Achieved Hypertension, diabetes, atherosclerosis, and hyperhomocysteinemia are among the most prevalent cardiovascular diseases (CVDs), severely affecting quality of life and healthcare costs. The identification of innovative pharmacological and nutraceutical strategies is therefore a major priority. In recent years, hydrogen sulfide (H₂S) has emerged as a key endogenous mediator in the cardiovascular system, and impaired H₂S biosynthesis has been associated with the development of several CVDs. Accordingly, natural sulfur-containing compounds, including polysulfides and isothiocyanates (ITCs), capable of releasing H₂S in an endogenous-like manner, represent promising candidates for cardiovascular prevention and therapy. Ferroptosis, a recently characterized iron-dependent form of regulated cell death driven by lipid peroxidation, has been increasingly implicated in the pathogenesis of CVDs. Although synthetic ferroptosis inhibitors such as ferrostatin-1 and deferoxamine have shown beneficial effects in preclinical studies, their clinical applicability is limited, particularly for chronic treatments. Since impaired endogenous H₂S production promotes ferroptosis, H₂S-releasing compounds may represent an innovative and safer strategy to counteract ferroptosis-mediated cardiovascular damage. This project investigated the H₂S-donor properties and anti-ferroptotic activity of selected natural sulfur-containing compounds, with particular focus on H₂S-releasing isothiocyanates (ITCs-H₂S donors). Cell-free assays and human aortic endothelial (HAECs) and smooth muscle cells (HASMCs) were used to characterize H₂S release kinetics and to evaluate their protective effects under ferroptotic and cardiovascular stress conditions induced by high glucose, oxidized LDL and homocysteine. The project achieved its main objectives. Cell-free and intracellular studies identified erucin and novel octyl-itaconate-based hybrid isothiocyanates as slow, thiol-dependent H₂S donors capable of efficiently releasing H₂S within vascular cells. These compounds displayed sustained intracellular H₂S generation in both endothelial and smooth muscle cells, supporting their further investigation in cardiovascular models. Experimental models demonstrated that erastin reliably induced ferroptosis in vascular cells, characterized by glutathione depletion, increased lipid peroxidation, reduced cell viability and modulation of canonical ferroptosis-related genes. Under high-glucose conditions, vascular cells developed oxidative stress and ferroptosis-like features, including glutathione depletion, enhanced lipid peroxidation and reduced viability. In contrast, oxidized LDL and homocysteine did not induce a consistent ferroptotic phenotype under the experimental conditions tested. Erucin significantly attenuated lipid peroxidation, restored intracellular glutathione levels, improved cell viability and modulated key ferroptosis-related targets, supporting its role as an effective H₂S donor able to counteract ferroptosis-like mechanisms associated with diabetic vascular injury. Based on these findings, erucin was selected for translational evaluation in a murine model of type 2 diabetes. Chronic oral administration improved glucose tolerance, preserved insulin secretion, and ameliorated motor performance. Molecular analyses demonstrated downregulation of pro-ferroptotic mediators (including ACSL4 and ALOX12) together with restoration of GPX4 and SLC7A11 expression in cardiovascular tissues. Histological analyses further revealed reduced inflammatory infiltration and improved tissue architecture in adipose and perivascular tissues of treated diabetic mice. Overall, the project provides preclinical evidence that slow H₂S-releasing isothiocyanates, particularly erucin, can modulate ferroptosis-related pathways and mitigate diabetes-associated vascular damage by restoring redox homeostasis and limiting lipid peroxidation. These findings support the hypothesis that H₂S-releasing compounds may represent promising candidates for the prevention and treatment of cardiovascular diseases, although further studies will be required to confirm their translational potential. Progetto finanziato nell’ambito del Piano Nazionale di Ripresa e Resilienza PNRR - Missione 4 – Componente 2 – Investimento 1.1 “Fondo per il Programma Nazionale di Ricerca e Progetti di Rilevante Interesse Nazionale (PRIN)” (Bando indetto con D.D. del MUR n. 104 del 02/02/2022).
Dettagli del progetto
Responsabile scientifico: Eleonora Turrini
Strutture Unibo coinvolte:
Dipartimento di Scienze per la Qualità della Vita
Coordinatore:
Università di PISA(Italy)
Contributo totale Unibo: Euro (EUR) 93.947,00
Durata del progetto in mesi: 24
Data di inizio
16/10/2023
Data di fine:
28/02/2026
