Fisetin Ameliorates Vascular Calcification by Regulating HNRNPA1-Mediated Ferroptosis.

Abstract

BACKGROUND: Fisetin has shown anti-inflammation, antioxidation, and antitumor effects, which could repress ferroptosis-related oxidative stress to attenuate the atherosclerotic progression. The current study aimed to further investigate the effect of fisetin on vascular calcification and its interaction with ferroptosis during the process. METHODS: Aortic smooth muscle cell (ASMC) was induced by high phosphate (HP) to mimic vascular calcification cellular model. Rats were treated by vitamin D3 and nicotine to mimic the vascular calcification animal model. 7.5 &#x3bc;M fisetin, 15 &#x3bc;M fisetin, ferrostatin-1 (Fer-1) (ferroptosis inhibitor), erastin (ferroptosis inducer), HNRNPA1 siRNA, were added to cellular model. Afterward, 100 mg/kg/day fisetin was administered in animal model (N = 3 in each group). RESULTS: Fisetin ameliorated calcification suggested by alizarin red relative level, BMP2 and RUNX2 expressions, and reduced ferroptosis indicated by reactive oxygen species level, mitochondrial membrane potential, cell viability, ACSL4, GPX4 and SLC7A11 expressions in a dose-dependent tendency in HP-treated ASMCs (all P < 0.05). Meanwhile, Fer-1 also reduced the calcification-related indexes in HP-treated ASMCs, but erastin promoted these indexes and compensated the effect of fisetin on these indexes (all P < 0.05). Regarding HNRNPA1, fisetin upregulated it in a dose-dependent manner and HNRNPA1 siRNA exhibited important function in attenuating the effect of fisetin on calcification-related indexes and ferroptosis-indexes in HP-treated ASMCs (all P < 0.05). In animal model, fisetin weakened ultrasound and HE staining reflected vascular calcification, lowered BMP2 and RUNX2 expressions, and upregulated GPX4, SLC7A11 and HNRNPA1 expressions (all P < 0.05). CONCLUSION: Fisetin ameliorates vascular calcification via regulating HNRNPA1-mediated ferroptosis under the cellular and rat environment in this preclinical study.