Protein Kinase D Knockdown Alleviates Vascular Calcification Through P53‐SLC7A11 Axis‐Mediated Ferroptosis

ABSTRACT Vascular calcification results in the hardening and weakening of the vascular wall, thereby increasing the risk of cardiovascular disease. Protein kinase D (PKD) is implicated in the regulation of numerous physiological processes, including cell growth, proliferation, differentiation, apoptosis, and intracellular signaling. PKD plays a pivotal role in regulating the function and phenotypic transformation of vascular smooth muscle cells (VSMCs); however, further studies are necessary to elucidate the role of PKD in vascular calcification. VSMCs were used to establish an in vitro calcification model. Six‐week‐old male C57BL/6J mice were selected to establish a vitamin D3 (Vitd3)‐induced calcification model in vivo. Adeno‐associated virus serotype 9 (AAV 9)‐PKD or AAV 9‐negative control was injected via tail vein 2 weeks before injecting Vitd3 into the mice. Protein expression levels of the P53‐SLC7A11 pathway were determined in both ex vivo and in vivo experiments. The degree of calcification was assessed by Alizarin red S (ARS) staining and calcium content measurement. We found that PKD was upregulated in ex vivo and in vivo models of vascular calcification. The inhibition of PKD resulted in a reduction in the expression of ferroptosis‐related genes and osteogenic differentiation of VSMCs, which in turn led to a mitigation of vascular calcification. Subsequent experiments demonstrated that the P53‐SLC7A11 signaling pathway is a key mediator of PKD‐induced vascular calcification. The results of this study indicate that PKD knockdown alleviates vascular calcification through the P53‐SLC7A11 axis‐mediated ferroptosis.