High Phosphate-Induced JAK-STAT Signalling Sustains Vascular Smooth Muscle Cell Inflammation and Limits Calcification

• Published in: Biomolecules (Basel, Switzerland) Vol. 14; no. 1; p. 29
• Main Authors: Macrì, Federica, Vigorito, Ilaria, Castiglione, Stefania, Faggiano, Stefano, Casaburo, Manuel, Fanotti, Nadia, Piacentini, Luca, Vigetti, Davide, Vinci, Maria Cristina, Raucci, Angela
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01.01.2024
• Subjects: Age; Analysis; Animals; Aorta; Atherosclerosis; Calcification; Calcification (ectopic); Calcium; Calcium phosphates; Cbfa-1 protein; Colorimetry; Enzyme-linked immunosorbent assay; Genetic aspects; Genotype & phenotype; Growth factors; Humans; hyperphosphataemia; Identification and classification; Immunohistochemistry; Inflammation; JAK-STAT; Janus kinase; Janus Kinases; Kinases; Methods; Mice; Monocyte chemoattractant protein 1; Muscle, Smooth, Vascular; Phosphatase; Phosphates; Proteins; Senescence; Signal Transduction; Smooth muscle; STAT Transcription Factors; Stat1 protein; vascular calcification; Vascular Calcification - chemically induced; vascular smooth muscle cells; Vitamin D; Western immunoblotting; Italy; Basel Switzerland; United States > US; Switzerland; JAK-STAT; inflammation; vascular smooth muscle cells; hyperphosphataemia; vascular calcification
• ISSN: 2218-273X; 2218-273X
• DOI: 10.3390/biom14010029

Vascular calcification (VC) is an age-related complication characterised by calcium-phosphate deposition in the arterial wall driven by the osteogenic transformation of vascular smooth muscle cells (VSMCs). The JAK-STAT pathway is an emerging target in inflammation. Considering the relationship between VC and inflammation, we investigated the role of JAK-STAT signalling during VSMC calcification. Human aortic smooth muscle cells (HASMCs) were cultured in high-inorganic phosphate (Pi) medium for up to 7 days; calcium deposition was determined via Alizarin staining and colorimetric assay. Inflammatory factor secretion was evaluated via ELISA and JAK-STAT members' activation using Western blot or immunohistochemistry on HASMCs or calcified aortas of Vitamin D-treated C57BL6/J mice, respectively. The JAK-STAT pathway was blocked by JAK Inhibitor I and Von Kossa staining was used for calcium deposits in murine aortic rings. During Pi-induced calcification, HASMCs released IL-6, IL-8, and MCP-1 and activated JAK1-JAK3 proteins and STAT1. Phospho-STAT1 was detected in murine calcified aortas. Blocking of the JAK-STAT cascade reduced HASMC proliferation and pro-inflammatory factor expression and release while increasing calcium deposition and osteogenic transcription factor RUNX2 expression. Consistently, JAK-STAT pathway inhibition exacerbates mouse aortic ring calcification ex vivo. Intriguingly, our results suggest an alternative link between VSMC inflammation and VC.