Jiedu Quyu Ziyin Decoction attenuates high-phosphate-induced calcification-associated phenotypes in cultured human aortic smooth muscle cells.

Abstract

Vascular calcification is one of the complications related to chronic kidney disease, and there are correlations between vascular calcification and oxidative stress, inflammation, and ferroptosis. This study examined whether Jiedu Quyu Ziyin Decoction (JQZYD) attenuates high-phosphate-induced calcification in cultured human aortic smooth muscle cells and evaluated the contribution of NRF2-associated antioxidant signalling. Human aortic vascular smooth muscle cells were exposed to high-phosphate conditions and treated with JQZYD. Calcification was assessed by Alizarin Red S staining, calcium content, alkaline phosphatase activity, and osteogenic markers. Ferroptosis-associated changes were evaluated by BODIPY 581/591 C11 oxidation, intracellular Fe2⁺, glutathione, glutathione peroxidase activity, and GPX4/SLC7A11 expression. NRF2 activation was assessed by immunofluorescence and protein analysis. Ferrostatin-1, RSL3, and NFE2L2 silencing were used for mechanistic evaluation. Inflammatory mediators and conditioned-medium effects were also examined. JQZYD reduced high-phosphate-induced mineral deposition, osteogenic marker expression, lipid peroxidation, Fe2⁺ accumulation, and inflammatory responses. These changes were accompanied by increased glutathione peroxidase activity, greater NRF2 nuclear accumulation, higher GPX4 and SLC7A11 transcript abundance, and increased NRF2 and GPX4 protein abundance. Ferrostatin-1 reproduced several protective effects of JQZYD. RSL3 and NFE2L2 silencing partially attenuated JQZYD-associated protection, supporting contributions of GPX4-related lipid peroxide regulation and NRF2-associated signalling without establishing direct NRF2 regulation of SLC7A11/GPX4 or pathway order. Conditioned medium from JQZYD-treated donor cells induced weaker inflammatory and calcific responses in recipient HASMCs, although the responsible mediators and potential contribution of residual treatment components remained unresolved. JQZYD attenuated high-phosphate-induced calcification-associated phenotypes and accompanying oxidative and inflammatory responses in cultured HASMCs. NFE2L2 silencing supports NRF2 involvement without establishing direct regulation of SLC7A11/GPX4 or pathway order. These findings are restricted to a cellular model and do not establish prevention of vascular calcification or therapeutic efficacy in CKD. Validation in CKD-relevant animal models and more physiologically complex vascular systems is required.

Key Findings

• Jiedu Quyu Ziyin Decoction (JQZYD) reduced high-phosphate-induced mineral deposition and osteogenic marker expression in human aortic smooth muscle cells.
• JQZYD decreased lipid peroxidation, Fe2⁺ accumulation, and inflammatory responses associated with vascular calcification.
• JQZYD treatment increased NRF2 nuclear accumulation, glutathione peroxidase activity, and expression of GPX4 and SLC7A11, key regulators of ferroptosis.
• Ferrostatin-1 mimicked protective effects of JQZYD, while RSL3 and NFE2L2 silencing partially reduced JQZYD's protective effects, indicating involvement of NRF2 and GPX4 in ferroptosis regulation.
• Conditioned medium from JQZYD-treated cells induced weaker inflammatory and calcific responses, suggesting paracrine effects.

Clinical Significance

Citation

Xiang Xiadan, Ran Ting, Guo Jiaoweiet al.. Jiedu Quyu Ziyin Decoction attenuates high-phosphate-induced calcification-associated phenotypes in cultured human aortic smooth muscle cells. Naunyn-Schmiedeberg's archives of pharmacology. 2026-Oct-24.