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C-type natriuretic peptide inhibiting vascular calcification might involve decreasing bone morphogenic protein 2 and osteopontin levels

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Abstract

Vascular calcification (VC) is highly associated with increased morbidity and mortality in patients with advanced chronic kidney disease. Paracrine/autocrine factors such as vasoactive peptides are involved in VC development. Here, we investigated the expression of the novel peptide C-type natriuretic peptide (CNP) in the vasculature, tested its ability to prevent VC in vivo and in vitro, and examined the mechanism involved. Rat aortic VC was induced by vitamin D3 plus nicotine (VDN). CNP (500 ng/kg/h) was administered by mini-osmotic pump. Calcification was examined by von Kossa staining; CNP and cyclic guanosine monophosphate (cGMP) contents were detected by radioimmunoassay, and mRNA and protein levels were examined by real-time PCR and Western blot analysis in aortas and calcified vascular smooth muscle cells (VSMCs). VDN-treated rat aortas showed higher CNP content and decreased expression of its receptor natriuretic peptide receptor B, along with increased vascular calcium deposition and alkaline phosphatase (ALP) activity. Low CNP levels were accompanied by increased vascular calcium deposition and ALP activity in VDN-treated rats when compared to vehicle treatment, which was further confirmed in cultured VSMCs. Administration of CNP greatly reduced VC in VDN-treated aortas compared with controls, which was confirmed in calcified VSMCs. The decrease in alpha-actin expression was ameliorated by CNP in vitro. Moreover, protein expression levels of osteopontin (OPN) were significantly up-regulated in calcified aortas, and CNP increased OPN expression in calcified aortas. Furthermore, CNP downregulated OPN and bone morphogenic protein 2 (BMP-2) expression in calcified aortas and VSMCs. Modulation of OPN and BMP-2 expression by CNP and the beneficial effects of CNP on calcified VSMCs were blocked significantly by protein kinase G inhibitor H7. Impaired local endogenous CNP and its receptor system may be associated with increased mineralization in vivo in rat aortas with VC, and administration of CNP inhibits VC development in vivo and in vitro, at least in part, via a cGMP/PKG pathway.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China to Jiang Z (Nos. 30570766, 30971169, 81170277) and research project from the Department of Education of Hunan Province (No. 05A043) and Aid Program for Science and Technology Innovative Research Team in Higher Educational Institutions of Human Province (2008-244) to Jiang Z. We are thankful to the Ministry of Education, P. R. China, and the University of South China for financial support to Jiang Z.

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Authors and Affiliations

  1. Key Laboratory for Arteriosclerology of Hunan Province, Institute of Cardiovascular Disease, University of South China, Hengyang, 421001, Hunan, People’s Republic of China

    Jing-Jing Chen, Ge-Bo Wen & Zhi-Sheng Jiang

  2. School of P.E. and Sports Science, Beijing Normal University, Beijing, People’s Republic of China

    Jing Zhang

  3. Key Laboratory of Molecular Cardiovascular Science, Ministry of Education, Peking University Health Science Center, Beijing, People’s Republic of China

    Yan Cai, Ye-Bo Zhou, Chao-Shu Tang & Yong-Fen Qi

  4. Jinan Fifth People’s Hospital, Jinan, Shandong, People’s Republic of China

    Jing-Jing Chen

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  1. Jing-Jing Chen
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Correspondence to Zhi-Sheng Jiang.

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Jing-Jing Chen and Jing Zhang have contributed equally to this work.

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Chen, JJ., Zhang, J., Cai, Y. et al. C-type natriuretic peptide inhibiting vascular calcification might involve decreasing bone morphogenic protein 2 and osteopontin levels. Mol Cell Biochem 392, 65–76 (2014). https://doi.org/10.1007/s11010-014-2019-1

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