Activation of Nuclear Factor Erythroid 2-Related Factor 2 Transcriptionally Upregulates Ectonucleotide Pyrophosphatase/Phosphodiesterase 1 Expression and Inhibits Ectopic Calcification in Mice

被引:1
作者
Ida, Tomomi [1 ]
Kanzaki, Hiroyuki [1 ]
Shimoyama, Miho [1 ]
Tohyama, Syunnosuke [1 ]
Ishikawa, Misao [2 ]
Katsumata, Yuta [1 ]
Arai, Chihiro [1 ]
Wada, Satoshi [3 ]
Manase, Shugo [1 ]
Tomonari, Hiroshi [1 ]
机构
[1] Tsurumi Univ, Sch Dent Med, Dept Orthodont, Yokohama, Kanagawa 2308501, Japan
[2] Tsurumi Univ, Sch Dent Med, Dept Anat, Yokohama, Kanagawa 2308501, Japan
[3] Kanazawa Med Univ, Dept Oral & Maxillofacial Surg, Kanazawa, Ishikawa 9201192, Japan
基金
日本学术振兴会;
关键词
ectopic calcification; Nrf2; pyrophosphate (PPi); osteoblastic differentiation; ENPP1; anti-oxidation; oxidative stress; INORGANIC PYROPHOSPHATE; OXIDATIVE STRESS; MINERALIZATION; BONE; CELLS; DIFFERENTIATION; IDENTIFICATION; OSSIFICATION; OSTEOBLASTS; PHOSPHATES;
D O I
10.3390/antiox13080896
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Calcification plays a key role in biological processes, and breakdown of the regulatory mechanism results in a pathological state such as ectopic calcification. We hypothesized that ENPP1, the enzyme that produces the calcification inhibitor pyrophosphate, is transcriptionally regulated by Nrf2, and that Nrf2 activation augments ENPP1 expression to inhibit ectopic calcification. Cell culture experiments were performed using mouse osteoblastic cell line MC3T3-E1. Nrf2 was activated by 5-aminolevulinic acid and sodium ferrous citrate. Nrf2 overexpression was induced by the transient transfection of an Nrf2 expression plasmid. ENPP1 expression was monitored by real-time RT-PCR. Because the promoter region of ENPP1 contains several Nrf2-binding sites, chromatin immunoprecipitation using an anti-Nrf2 antibody followed by real-time PCR (ChIP-qPCR) was performed. The relationship between Nrf2 activation and osteoblastic differentiation was examined by alkaline phosphatase (ALP) and Alizarin red staining. We used mice with a hypomorphic mutation in ENPP1 (ttw mice) to analyze whether Nrf2 activation inhibits ectopic calcification. Nrf2 and Nrf2 overexpression augmented ENPP1 expression and inhibited osteoblastic differentiation, as indicated by ALP expression and calcium deposits. ChIP-qPCR showed that some putative Nrf2-binding sites in the ENPP1 promoter region were bound by Nrf2. Nrf2 activation inhibited ectopic calcification in mice. ENPP1 gene expression was transcriptionally regulated by Nrf2, and Nrf2 activation augmented ENPP1 expression, leading to the attenuation of osteoblastic differentiation and ectopic calcification in vitro and in vivo. Nrf2 activation has a therapeutic potential for preventing ectopic calcification.
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页数:16
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共 58 条
[1]   Pyrophosphate inhibits mineralization of osteoblast cultures by binding to mineral, up-regulating osteopontin, and inhibiting alkaline phosphatase activity [J].
Addison, William N. ;
Azari, Fereshteh ;
Sorensen, Esben S. ;
Kaartinen, Mari T. ;
McKee, Marc D. .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2007, 282 (21) :15872-15883
[2]   ENPP1-Fc prevents mortality and vascular calcifications in rodent model of generalized arterial calcification of infancy [J].
Albright, Ronald A. ;
Stabach, Paul ;
Cao, Wenxiang ;
Kavanagh, Dillon ;
Mullen, Isabelle ;
Braddock, Alexander A. ;
Covo, Mariel S. ;
Tehan, Martin ;
Yang, Guangxiao ;
Cheng, Zhiliang ;
Bouchard, Keith ;
Yu, Zhao-Xue ;
Thorn, Stephanie ;
Wang, Xiangning ;
Folta-Stogniew, Ewa J. ;
Negrete, Alejandro ;
Sinusas, Albert J. ;
Shiloach, Joseph ;
Zubal, George ;
Madri, Joseph A. ;
De La Cruz, Enrique M. ;
Braddock, Demetrios T. .
NATURE COMMUNICATIONS, 2015, 6
[3]   COMPARISON OF NORMAL AND RACHITIC RAT MATRIX VESICLES [J].
ANDERSON, HC ;
STECHSCHULTE, DJ ;
HSU, HHT ;
MORRIS, DC .
BONE AND MINERAL, 1992, 17 (02) :119-122
[4]   Role of inorganic polyphosphate in mammalian cells: from signal transduction and mitochondrial metabolism to cell death [J].
Angelova, Plamena R. ;
Baev, Artyom Y. ;
Berezhnov, Alexey V. ;
Abramov, Andrey Y. .
BIOCHEMICAL SOCIETY TRANSACTIONS, 2016, 44 :40-45
[5]   The Neuropeptide Spexin Promotes the Osteoblast Differentiation of MC3T3-E1 Cells via the MEK/ERK Pathway and Bone Regeneration in a Mouse Calvarial Defect Model [J].
Assefa, Freshet ;
Kim, Ju Ang ;
Lim, Jiwon ;
Nam, Sang-Hyeon ;
Shin, Hong-In ;
Park, Eui Kyun .
TISSUE ENGINEERING AND REGENERATIVE MEDICINE, 2022, 19 (01) :189-202
[6]   CALCIFICATION MECHANISMS - ROLES FOR CELLS AND MINERAL [J].
BECKER, GL .
JOURNAL OF ORAL PATHOLOGY & MEDICINE, 1977, 6 (05) :307-315
[7]   INORGANIC-PHOSPHATE ADDED EXOGENOUSLY OR RELEASED FROM BETA-GLYCEROPHOSPHATE INITIATES MINERALIZATION OF OSTEOID NODULES INVITRO [J].
BELLOWS, CG ;
HEERSCHE, JNM ;
AUBIN, JE .
BONE AND MINERAL, 1992, 17 (01) :15-29
[8]   The Nrf2-Inducible Antioxidant Defense in Astrocytes can be Both Up- and Down-Regulated by Activated Microglia: Involvement of p38 MAPK [J].
Correa, Fernando ;
Ljunggren, Elin ;
Mallard, Carina ;
Nilsson, Michael ;
Weber, Stephen G. ;
Sandberg, Mats .
GLIA, 2011, 59 (05) :785-799
[9]   Use of parathyroid hormone in hypoparathyroidism [J].
Cusano, N. E. ;
Rubin, M. R. ;
Irani, D. ;
Sliney, J., Jr. ;
Bilezikian, J. P. .
JOURNAL OF ENDOCRINOLOGICAL INVESTIGATION, 2013, 36 (11) :1121-1127
[10]   Calcinosis in systemic sclerosis [J].
Davuluri, Srijana ;
Lood, Christian ;
Chung, Lorinda .
CURRENT OPINION IN RHEUMATOLOGY, 2022, 34 (06) :319-327