Melatonin Inhibits NF-κB/CREB/Runx2 Signaling and Alleviates Aortic Valve Calcification

Abstract

Calcific aortic valve disease (CAVD) is linked to high mortality. Melatonin inhibits nuclear factor-kappa B (NF-κB)/cyclic AMP response element-binding protein (CREB), contributing to CAVD progression. This study determined the role of melatonin/MT1/MT2 signaling in valvular interstitial cell (VIC) calcification. Western blotting and Alizarin red staining were used to analyze NF-κB/CREB/runt-related transcription factor 2 (Runx2) signaling in porcine VICs treated with an osteogenic (OST) medium without (control) or with melatonin for 5 days. Chromatin immunoprecipitation (ChIP) assay was used to analyze NF-κB's transcription regulation of NF-κB on the Runx2 promoter. OST medium-treated VICs exhibited a greater expression of NF-κB, CREB, and Runx2 than control VICs. Melatonin treatment downregulated the effects of the OST medium and reduced VIC calcification. The MT1/MT2 antagonist (Luzindole) and MT1 receptor neutralized antibody blocked the anticalcification effect of melatonin, but an MT2-specific inhibitor (4-P-PDOT) did not. Besides, the NF-κB inhibitor (SC75741) reduced OST medium-induced VIC calcification to a similar extent to melatonin at 10 nmol/L. The ChIP assay demonstrated that melatonin attenuated OST media increased NF-κB binding activity to the promoter region of Runx2. Activation of the melatonin/MT1-axis significantly reduced VIC calcification by targeting the NF-κB/CREB/Runx2 pathway. Targeting melatonin/MT1 signaling may be a potential therapeutic strategy for CAVD.

Targeted Radiation Exposure Induces Accelerated Aortic Valve Remodeling in ApoE−/− Mice

“…Recently, Candellier et al demonstrated that indoxyl-sulfate promotes osteogenic differentiation of hVIC via an activation of the AhR-NF-κB pathway [ 29 ]. Our preclinical model could be used to evaluate the impact of therapies inhibiting NF-κB signaling, including melatonin [ 31 ], aspirin [ 32 ], or metformin [ 33 ], on radiation-induced aortic remodeling.…”

The Role and Therapeutic Potential of Melatonin in Degenerative Fundus Diseases: Diabetes Retinopathy and Age-Related Macular Degeneration

“…121 As well as blocking NF-κB activation signaling, melatonin inhibits the activity of IKK receptors. 122 Furthermore, melatonin regulates cytokine signaling pathways, including the Janus kinase/signal transducer and activator of transcription (JAK/STAT) family pathway, impacting immune cell function and the development of inflammatory responses. 123 These signaling pathways play an important role in the inflammatory response, and melatonin inhibits the onset and progression of the inflammatory response by regulating their activity.…”

“… 121 As well as blocking NF-κB activation signaling, melatonin inhibits the activity of IKK receptors. 122 Furthermore, melatonin regulates cytokine signaling pathways, including the Janus kinase/signal transducer and activator of transcription (JAK/STAT) family pathway, impacting immune cell function and the development of inflammatory responses. 123 …”

The Complex Relationship between Hypoxia Signaling, Mitochondrial Dysfunction and Inflammation in Calcific Aortic Valve Disease: Insights from the Molecular Mechanisms to Therapeutic Approaches

“…Hence, one of the causes of ferroptosis is an iron overload that is involved in CAVS by enhancing calcium deposition and calcification in endothelial cells (HUVEC) [ 90 ]. It is identified that the endothelial cells play crucial impacts in the calcification process through the EndMT [ 91 ]. Multiple pathways are involved in this process including the signaling pathways involved in hypoxia and inflammation, such as the transforming growth factor beta (TGF-β) signaling pathway [ 92 ], and the Wnt signaling pathway [ 93 , 94 ].…”