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Activation of canonical Wnt/β-catenin signaling inhibits H2O2-induced decreases in proliferation and differentiation of human periodontal ligament fibroblasts

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Abstract

Human periodontal ligament fibroblasts (hPLFs) are exposed to oxidative stress during periodontal inflammation and dental treatments. It is hypothesized that hydrogen peroxide (H2O2)-mediated oxidative stress decreases survival and osteogenic differentiation of hPLFs, whereas these decreases are prevented by activation of the Wnt pathway. However, there has been a lack of reports that define the exact roles of canonical Wnt/β-catenin signaling in H2O2-exposed hPLFs. Treatment with H2O2 reduced viability and proliferation in hPLFs in a dose- and time-dependent manner and led to mitochondria-mediated apoptosis. Pretreatment with lithium chloride (LiCl) or Wnt1 inhibited the oxidative damage that occurred in H2O2-exposed hPLFs. However, knockout of β-catenin or treatment with DKK1 facilitated the H2O2-induced decreases in viability, mitochondrial membrane potential, and Bcl-2 induction. Osteoblastic differentiation of hPLFs was also inhibited by combined treatment with 100 μM H2O2, as evidenced by the decreases in alkaline phosphatase (ALP) activity and mineralization. H2O2-mediated inhibition of osteoblast differentiation in hPLFs was significantly attenuated in the presence of 500 ng/ml Wnt1 or 20 mM LiCl. In particular, H2O2 stimulated the expression of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) at protein and mRNA levels in hPLFs, whereas the induction was almost completely suppressed in the presence of Wnt1 or LiCl. Furthermore, siRNA-mediated silencing of Nrf2 blocked H2O2-induced decreases in ALP activity and mineralization of hPLFs with the concomitant restoration of runt-related transcription factor 2 and osteocalcin mRNA expression and ALP activity. Collectively, these results suggest that activation of the Wnt/β-catenin pathway improves proliferation and mineralization in H2O2-exposed hPLFs by downregulating Nrf2.

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Acknowledgments

This research was supported by Basic Science Research Program through the National Research Foundation of Korea (KRF) funded by the Ministry of Science, ICT and future Planning (NRF-2013R1A2A2A01967207).

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    Authors and Affiliations

    1. Cluster for Craniofacial Development & Regeneration Research, Chonbuk National University, Jeonju, 561-756, South Korea

      Sung-Ho Kook, Eui-Sic Cho, Sher Bahadur Poudel, Yu-Hyeon Ahn, Jae-Won Hwang & Jeong-Chae Lee

    2. Department of Bioactive Material Sciences, Institute of Oral Biosciences (BK21 program) and School of Dentistry, Chonbuk National University, Jeonju, 561-756, South Korea

      Sung-Ho Kook, Daewoo Lee, Eui-Sic Cho, Sher Bahadur Poudel, Yu-Hyeon Ahn, Jae-Won Hwang, Hyeok Ji, Jong-Ghee Kim & Jeong-Chae Lee

    3. Department of Maxillofacial Biomedical Engineering and Institute of Oral Biology, School of Dentistry, Kyung Hee University, Seoul, 130-701, South Korea

      Jung Sun Heo

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    1. Sung-Ho Kook
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    Sung-Ho Kook and Daewoo Lee have contributed equally to this work.

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    Kook, SH., Lee, D., Cho, ES. et al. Activation of canonical Wnt/β-catenin signaling inhibits H2O2-induced decreases in proliferation and differentiation of human periodontal ligament fibroblasts. Mol Cell Biochem 411, 83–94 (2016). https://doi.org/10.1007/s11010-015-2570-4

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