Purpose of Review: Vascular calcification (VC) is increasingly recognized as an actively regulated pathological process rather than passive mineral deposition, strongly associated with aging, atherosclerosis, and chronic kidney disease. Sclerostin, a Wnt signalling antagonist primarily expressed in osteocytes, has recently been implicated in VC, although its precise vascular role remains debated. This review aims to integrate mechanistic and translational evidence on how hypoxia and hypoxia-inducible factor-1α (HIF-1α) regulate sclerostin and contribute to vascular mineralisation. Recent Findings: Experimental studies demonstrate that HIF-1α activation links hypoxia to key osteogenic pathways, including BMP2, RUNX2, and Wnt/β-catenin signalling, thereby influencing phenotypic switching of vascular smooth muscle cells (VSMCs). Hypoxia exerts both stimulatory and suppressive effects on sclerostin depending on local tissue conditions, reflecting a context-dependent regulatory network. Preclinical and clinical data show that sclerostin can act as either a compensatory inhibitor or a pro-calcific mediator, depending on disease stage and metabolic environment. Emerging evidence further highlights natural products such as polyphenols, flavonoids, and marine-derived compounds that modulate sclerostin expression through oxidative, inflammatory, and Wnt-related pathways. Summary: Sclerostin sits at a critical intersection between bone and vascular systems, where hypoxia-driven HIF signalling orchestrates its dual effects on mineral metabolism. Understanding the HIF–sclerostin axis provides new insight into the bone–vascular continuum and identifies potential therapeutic targets. Natural bioactive compounds capable of restoring sclerostin–Wnt balance may represent safe, multi-targeted strategies to mitigate VC progression, warranting further mechanistic and translational evaluation. Graphical

Background. Sailuotong (SLT) is a standardized three‐herb formulation consisting of extracts of Panax ginseng, Ginkgo biloba, and Crocus sativus for the treatment of vascular dementia (VaD). Although SLT has been shown to increase cerebral blood flow, the direct effects of SLT on vascular reactivity have not been explored. This study aims to examine the vasodilatory effects of SLT and the underlying mechanisms in rat isolated tail artery. Methods. Male (250–300 g) Wistar Kyoto (WKY) rat tail artery was isolated for isometric tension measurement. The effects of SLT on the influx of calcium through the cell membrane calcium channels were determined in Ca²⁺‐free solution experiments. Results. SLT (0.1–5,000 μg/ml) caused a concentration‐dependent relaxation in rat isolated tail artery precontracted by phenylephrine. In the contraction experiments, SLT (500, 1,000, and 5,000 μg/mL) significantly inhibited phenylephrine (0.001 to 10 μM)‐ and KCl (10–80 mM)‐induced contraction, in a concentration‐dependent manner. In Ca²⁺‐free solution, SLT (500, 1,000, and 5,000 μg/mL) markedly suppressed Ca²⁺‐induced (0.001–3 mM) vasoconstriction in a concentration‐dependent manner in both phenylephrine (10 μM) or KCl (80 mM) stimulated tail arteries. L‐type calcium channel blocker nifedipine (10 μM) inhibited PE‐induced contraction. Furthermore, SLT significantly reduced phenylephrine‐induced transient vasoconstriction in the rat isolated tail artery. Conclusion. SLT induces relaxation of rat isolated tail artery through endothelium‐independent mechanisms. The SLT‐induced vasodilatation appeared to be jointly meditated by blockages of extracellular Ca²⁺ influx via receptor‐gated and voltage‐gated Ca²⁺ channels and inhibition of the release of Ca²⁺ from the sarcoplasmic reticulum.

Background: Vascular calcification (VC) is a pathological process associated with aging, atherosclerosis, metabolic syndrome, and chronic kidney disease. It involves vascular smooth muscle cells (VSMCs) transforming into osteoblast-like cells, leading to mineral deposition within the vascular wall. Transcription factors such as runt-related transcription factor 2 (RUNX2) and bone matrix-associated proteins, including sclerostin (SOST), osteoprotegerin, and osteopontin, play crucial roles in this process. Cannabidiol (CBD), a cannabis-derived compound, has demonstrated cardiovascular benefits, but its impact on VC remains unclear. Aims: This study aimed to (1) develop a human aortic smooth muscle cell (HASMC) VC model to investigate the role of bone-related proteins in calcification, (2) analyze protein expression changes in the VC model using proteomic techniques, and (3) evaluate the effects of CBD on the osteogenic transition of HASMCs under calcifying conditions. Methods: HASMCs were cultured under calcifying conditions with inorganic phosphate and calcium to induce VC. Calcification was quantified using Alizarin Red S staining, while Western blotting and enzymatic assays assessed bone-related protein expression, including alkaline phosphatase and RUNX2. Proteomic analysis identified dysregulated pathways and proteins associated with VC. CBD cytotoxicity was evaluated at concentrations up to 8 µM, and apoptosis assays were performed in both calcifying and non-calcifying conditions. Results: Calcifying conditions induced significant phenotypic changes in HASMCs, characterized by reduced smooth muscle marker expression and calcium nodule formation. Enzymatic assays revealed increased alkaline phosphatase activity, while proteomic analysis identified upregulation of cardiovascular-related proteins, including smoothelin, succinate dehydrogenase complex flavoprotein subunit A, and Rho GTPase–activating protein 17. CBD treatment unexpectedly increased calcium deposits and apoptotic activity in calcified HASMCs, suggesting a potentially adverse role in the calcification process. Conclusion: Elevated phosphate levels drive osteogenic changes in HASMCs, contributing to VC progression. While CBD has shown promise in cardiovascular therapy, its exacerbation of calcium deposition in this model highlights the need for caution. These findings underscore the importance of understanding VC mechanisms and the therapeutic implications of CBD in vascular health.

Jing Guan Fang (JGF) is a traditional multi-herb formula used for hyperinflammatory conditions associated with severe viral infections; however, its protein-level effects in macrophage-mediated inflammation remain incompletely characterised. In this study, we combined ultra-performance liquid chromatography–quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS)-based phytochemical profiling, functional anti-inflammatory assays, and discovery-level proteomic and secretome analyses to investigate the anti-inflammatory effects of JGF in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Phytochemical profiling revealed predominantly flavonoid- and phenolic-related annotated features, together with selected terpenoid- and alkaloid-related annotations. Functionally, JGF treatment was associated with reduced nitric oxide production and differential effects on pro-inflammatory cytokine release, with a clearer concentration-dependent reduction in interleukin-6 (IL-6) and a more variable tumour necrosis factor-α (TNF-α) response, without reducing cell viability. Label-free quantitative proteomics suggested that JGF treatment was associated with modulation of LPS-induced protein changes, including reduced abundance of inflammation-associated proteins such as myristoylated alanine-rich C-kinase substrate (MARCKS) and inducible nitric oxide synthase (NOS2), together with partial restoration of selected regulatory proteins such as AIMP1 and AIMP2. Secretome analysis further suggested that JGF reduced extracellular inflammatory and tissue-remodelling-related mediators, including SERPINE1/PAI-1. Exploratory pathway analysis indicated that JGF treatment was associated with changes in inflammation-related and oxidative stress-related signalling networks. Collectively, these findings provide discovery-level molecular insights into the anti-inflammatory effects of JGF in LPS-stimulated macrophages and support further targeted validation of its pharmacological activity as a candidate multi-component anti-inflammatory formula.