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Role of reactive oxygen species in angiotensin II: induced receptor activator of nuclear factor-κB ligand expression in mouse osteoblastic cells

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Abstract

Angiotensin II (Ang II) has been shown to induce receptor activator of nuclear factor-κB ligand (RANKL) expression in osteoblasts associated with its effect on reactive oxygen species (ROS) production. The objective of the present study was to investigate the potential pathways by which Ang II induces RANKL expression and the role of ROS in Ang II-induced RANKL expression in mouse osteoblastic MC3T3-E1 cells. Treatment with Ang IIinduced RANKL expression in a dose- and time-dependent manner in osteoblasts, which was attenuated by pre-treatment with an AT1 receptor antagonist (olmesartan), ROS scavenger (N-acetylcysteine, NAC), or the ERK inhibitor (U0126), but not with AT2R antagonist (PD123319). Furthermore, Ang II enhanced AT1R and NAD(P)H oxidase (NOX) p22phox and p67phox expression and activity in osteoblasts. In addition, Ang II promoted ROS production, which was mitigated by pre-treatment with olmesartan or a NOX inhibitor (diphenyleneiodonium, DPI), but not with PD1123319 or U0126, in osteoblasts. Moreover, Ang II enhanced the ERK1/2 phosphorylation, which was abrogated by pre-treatment with olmesartan, NAC, DPI, or U0126 in osteoblasts. These results suggest that Ang II, through its AT1R, enhanced NOX activity and ROS production, and activated the ERK pathway to up-regulate RANKL expression in osteoblasts in vitro.

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Authors and Affiliations

  1. Department of Orthopaedics, The Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an, 710004, Shaanxi, China

    Yanqiu Zhang, Yongtao Zhang, Chunsheng Wang & Kunzheng Wang

  2. Department of Physical Education, Xi’an Shiyou University, Xi’an, 710065, Shaanxi, China

    Yanqiu Zhang

  3. Department of Orthopedics, The Affiliated Hospital of Qingdao University, Qingdao, 26000, Shandong, China

    Yongtao Zhang & Jianqiang Kou

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  1. Yanqiu Zhang
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  2. Yongtao Zhang
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  3. Jianqiang Kou
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  4. Chunsheng Wang
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  5. Kunzheng Wang
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Correspondence to Kunzheng Wang.

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Supplementary Fig. 1

Intracellular ROS levels in cells in different groups were detected by fluorescence microscopy. Supplementary material 1 (TIFF 10,879 kb).

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Zhang, Y., Zhang, Y., Kou, J. et al. Role of reactive oxygen species in angiotensin II: induced receptor activator of nuclear factor-κB ligand expression in mouse osteoblastic cells. Mol Cell Biochem 396, 249–255 (2014). https://doi.org/10.1007/s11010-014-2160-x

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  • DOI: https://doi.org/10.1007/s11010-014-2160-x

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