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Updated: Aug 28, 2026

Isolation of Primary Patient-specific Aortic Smooth Muscle Cells and Semiquantitative Real-time Contraction Measurements In Vitro
Published on: February 15, 2022
Relaxin-3/RXFP1 Axis Regulates the Phenotypic Switch of Diabetic Vascular Smooth Muscle Cells by Inhibiting
Jiaxin Xue1, Jingzhi Wang1, Yuxin Yang1
1The Department of Cardiology, The First Affiliated Hospital of Harbin Medical University, China (J.X., J.W., Y.Y., X. Liang, X. Liu, S.H., X.Z.).
Background:
The phenotypic switching of vascular smooth muscle cells (VSMCs) is a critical pathophysiological mechanism of diabetic vascular remodeling. There is increasing evidence indicating that endogenous relaxin-3 exhibits cardiovascular effects, including vasodilation, reversal of ventricular remodeling, and attenuation of myocardial fibrosis. This study investigates the role of relaxin-3 as a key regulator in the phenotypic switching of VSMCs and elucidates its underlying mechanisms.
Methods:
We generated a diabetic rat model to investigate the biological roles of relaxin-3 in diabetic vascular remodeling. Metabolomic profiling characterized relaxin-3-induced metabolic changes in VSMCs under high glucose treatment. Combined chromatin immunoprecipitation sequencing and RNA sequencing data were used to further investigate the gene regulatory mechanisms of relaxin-3 in the phenotypic switching of VSMCs and target genes regulated by H3K18la (histone H3 lysine 18 lactylation).
Results:
Relaxin-3 and its receptor RXFP1 (relaxin family peptide receptor 1) were upregulated in diabetic human and rat aortas. Silencing of relaxin-3 or RXFP1 expression inhibited VSMC contractile phenotype protein expression and promoted VSMC proliferation. Relaxin-3 treatment mitigated the phenotypic switching of VSMCs via RXFP1, preserving VSMCs' contractile phenotype. Further, relaxin-3 treatment lowered the glycolytic rate and lactate production during the phenotypic switching of VSMCs and decreased histone lactylation. Mechanistically, relaxin-3 suppressed LDHA (lactate dehydrogenase A)-mediated lactate production and H3K18la-mediated EGFR (epidermal growth factor receptor) transcription, and blocked PKM2 (pyruvate kinase M2) nuclear translocation, thereby preventing PKM2/β-catenin complex formation.
Conclusions:
Relaxin-3 alleviated the phenotypic switching of diabetic VSMCs through a dual mechanism: inhibiting lactate-mediated H3K18la and disrupting the intranuclear PKM2/β-catenin interaction, thereby preserving the contractile VSMC phenotype.
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