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Hops extract and xanthohumol ameliorate bone loss induced by iron overload via activating Akt/GSK3β/Nrf2 pathway

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Journal of Bone and Mineral Metabolism Aims and scope Submit manuscript

Abstract

Introduction

Osteoporosis is closely related to iron metabolism. This study aimed to investigate whether hops extract (HLE) and its active component xanthohumol (XAN) could ameliorate bone loss caused by iron overload, and explored its potential mechanism.

Materials and methods

Iron overload mice induced by iron dextran (ID) were used in vivo, and were treated with HLE and XAN for 3 months. Bone micro-structure and bone morphology parameters were determined by Micro-CT and TRAP staining. Bone metabolism markers and oxidation indexes in serum and bone tissue were evaluated. For in vitro experiment, bone formation indexes were determined. Moreover, the expression of key proteins in protein kinase B (Akt)/glycogen synthetase kinase 3β (GSK3β)/nuclear factor E2-related (Nrf2) pathway was evaluated by Western blotting.

Results

HLE and XAN effectively improved the bone micro-structure of the femur in mice, altered bone metabolism biomarkers, and regulated the expression of proteins related to bone metabolism. Additionally, they significantly promoted cell proliferation, runt-related gene 2 (Runx2) expression, and increased ALP activity in ID-induced osteoblasts. Moreover, HLE and XAN markedly inhibited the increase of oxidative stress caused by iron overload in vivo and in vitro. Further studies showed that they significantly up-regulated the expression of p-Akt, p-GSK3β, nuclear-Nrf2, NAD(P)H: quinone oxidoreductase 1 (NQO1), and heme oxygenase-1 (HO-1) in ID-induced osteoblasts.

Conclusion

These findings indicated hops and xanthohumol could ameliorate bone loss induced by iron overload via activating Akt/GSK3β/Nrf2 pathway, which brought up a novel sight for senile osteoporosis therapy.

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Data availability

All data generated or analyzed during this study are included in this published article.

Abbreviations

HLE:

Humulus lupulus Extract

XAN:

Xanthohumol

NAC:

N-Acety-l-cysteine

ELISA:

Enzyme-linked immunosorbent assay

RANKL:

Receptor activator of nuclear factor-κB ligand

OPG:

Osteoclastogenesis

SOD:

Superoxide dismutase

ALP:

Alkaline phosphatase

Akt:

Protein kinase B

GSK3β:

Glycogen synthetase kinase 3β

Nrf2:

Nuclear factor E2-related

NQO1:

NAD(P)H: quinone oxidoreductase 1

HO-1:

Heme oxygenase-1

Runx2:

Runt-related gene 2

COL-I:

Collagen I

NFATC1:

Nuclear factor of activated T cells 1

MMP9:

Matrix metalloproteinase 9

CtsK:

Cathepsin K

CTX-I:

C-terminal telopeptides of type I collagen

TRACP-5b:

Tartrate-resistant acid phosphatase 5b

ID:

Iron dextran

MDA:

Malondialdehyde

GSH-PX:

Glutathione peroxidase

ROS:

Reactive oxygen species

PMSF:

Phenylmethylsulfonyl fluoride

SDS-PAGE:

Sodium dodecylsulfate olyacrylamide gel electrophoresis

ECL:

Enhanced chemiluminescent

SEM:

Standard error of the mean

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Funding

This work was supported by the Natural Science Foundation of China (Nos. 82174079, U1603283, 82004015) and Science and Technology Commission of Shanghai Municipality (21S21902600).

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Author notes

  1. Xiaolei Sun, Tianshuang Xia and Shiyao Zhang contributed equally to the work.

Authors and Affiliations

  1. Department of Pharmacognosy, School of Pharmacy, Naval Medical University, 325 Guohe Road, Shanghai, 200433, China

    Xiaolei Sun, Tianshuang Xia, Jiabao Zhang, Ting Han & Hailiang Xin

  2. School of Pharmacy, Shandong University of Traditional Chinese Medicine, 4655 Daxue Road, Jinan, 250355, China

    Xiaolei Sun, Shiyao Zhang & Lingchuan Xu

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  1. Xiaolei Sun
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  6. Ting Han
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Contributions

HLX, XLS and TSX designed the experiments; XLS and SYZ performed the experiments; XLS and TSX analyzed the data and wrote the manuscript; TSX and JBZ helped to organize figures; LCX, TH and HLX reviewed the manuscript.

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Correspondence to Lingchuan Xu, Ting Han or Hailiang Xin.

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Sun, X., Xia, T., Zhang, S. et al. Hops extract and xanthohumol ameliorate bone loss induced by iron overload via activating Akt/GSK3β/Nrf2 pathway. J Bone Miner Metab 40, 375–388 (2022). https://doi.org/10.1007/s00774-021-01295-2

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