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Hyperoside prevents osteoporosis by activating the PI3K/AKT signaling pathway to inhibit oxidative stress and promote osteogenesis

  • Authors:
    • Lirong Li
    • Huaidong Deng
    • Shabin Zhuan
    • Boyu Wu
    • Dawei Xiao
  • View Affiliations / Copyright

    Affiliations: Department of Orthopedics, Dongguan Traditional Chinese Medicine Hospital, Dongguan, Guangdong 523000, P.R. China, Department of Orthopedics, Dongguan Traditional Chinese Medicine Hospital, Dongguan, Guangdong 523000, P.R. China
    Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 57
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    Published online on: December 3, 2025
       https://doi.org/10.3892/mmr.2025.13767
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Abstract

Hyperoside (Hyp), a naturally occurring flavonol glycoside derived from Crataegus, otherwise known as hawthorn, possesses potent antioxidant properties and has demonstrated therapeutic potential in various oxidative stress‑related diseases, including osteoporosis (OP). However, the precise molecular mechanisms underlying the anti‑osteoporotic effects of Hyp remain to be fully elucidated. The present study aimed to evaluate the therapeutic efficacy of Hyp against OP and to elucidate its underlying mechanisms. An osteoporotic mouse model was established via bilateral ovariectomy (OVX) to assess the in vivo efficacy of Hyp. Network pharmacology was employed to predict the potential therapeutic targets of Hyp in OP. In vitro experiments using bone marrow mesenchymal stem cells (BMSCs) were performed to validate the findings. Techniques including alkaline phosphatase staining, Alizarin red S staining, reverse transcription‑quantitative PCR and western blotting were used to assess osteogenic differentiation and molecular signaling pathways. Micro‑CT analysis revealed that Hyp significantly ameliorated OVX‑induced bone loss in mice. Network pharmacology identified the PI3K/AKT signaling pathway as a potential key target. In vitro, Hyp significantly reduced H2O2‑induced oxidative stress in BMSCs and promoted their osteogenic differentiation. Mechanistically, Hyp was found to activate the PI3K/AKT signaling pathway, suggesting its notable role in mediating the antioxidant and osteoinductive effects of Hyp. Summarily, Hyp may effectively alleviate OVX‑induced OP in mice, potentially by mitigating oxidative stress and promoting osteogenesis via activation of the PI3K/AKT signaling pathway. These findings provide novel insights into the therapeutic mechanism of Hyp and support its potential as a candidate agent for OP treatment.
View Figures

Figure 1

Hyp alleviates ovariectomy-induced
osteoporosis in mice. (A) Representative micro-CT three-dimensional
reconstruction images of the mouse femur. Quantitative analyses of
(B) Tb.Sp, (C) Tb.N, (D) Tb.Th and (E) BV/TV (n=5). (F)
Representative H&E staining images of femoral sections (n=5;
×20 magnification). (G) Representative TRAP-stained sections
showing osteoclasts (indicated by black arrows) (n=5; ×40
magnification). (H) Quantification of osteoblast number per bone
surface (n=5). (I) Quantification of osteoclast number per bone
surface (n=5). Data are presented as the mean ± standard deviation.
*P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001. Tb.Sp,
trabecular separation; Tb.N, trabecular number; Tb.Th, trabecular
thickness; BV/TV, bone volume fraction; TRAP, tartrate-resistant
acid phosphatase; Hyp, hyperoside.

Figure 2

Potential mechanism of Hyperoside for
the treatment of OP. (A) Volcano plots of the GSE35956 and GSE80614
gene chip datasets. (B) Venn diagram of disease and drug targets.
(C) Protein-protein interaction network constructed using the
STRING database of Hyperoside in the treatment of OP. (D)
Visualization and analysis of the PPI network performed using
Cytoscape software. (E) Bubble chart of Kyoto Encyclopedia of Genes
and Genomes enrichment analysis. (F) GO enrichment analysis. All
displayed GO terms met the significance threshold of adjusted
P<0.05. OP, osteoporosis; GO, Gene Ontology; BP, biological
process; CC, cellular component; MF, molecular function.

Figure 3

Hyp alleviates the inhibitory effect
of H2O2 on the viability of BMSCs. (A)
Chemical structure of Hyp. (B) Impact of Hyp on the viability of
BMSCs (n=3). (C) Impact of H2O2 on the
viability of BMSCs (n=3). (D) Impact of Hyp on the viability of
BMSCs under the intervention of H2O2 (n=3).
(E) Representative EDU staining images showing cell proliferation
in BMSCs treated with Hyp and H2O2 (n=3; ×5
magnification). Data are presented as the mean ± standard
deviation. *P<0.05, **P<0.01 and ****P<0.0001. BMSC, bone
marrow mesenchymal stem cells; Hyp, hyperoside.

Figure 4

Hyperoside alleviates the oxidative
stress of bone marrow mesenchymal stem cells induced by
H2O2. (A) Representative fluorescence images
of DCFH-DA staining (n=3; ×10 magnification). (B) Representative
flow cytometry images of DCFH-DA staining (n=3). (C) Quantification
of the fluorescence intensity of DCFH-DA staining and (D)
quantification of the mean fluorescence intensity in flow cytometry
(n=3). Quantification of the (E) MDA and (F) SOD levels (n=3). Data
are presented as the mean ± standard deviation. **P<0.01,
***P<0.001 and ****P<0.0001. Hyp, hyperoside; ROS, reactive
oxygen species; MDA, malondialdehyde; SOD, superoxide
dismutase.

Figure 5

Hyperoside rescues the osteogenic
differentiation function of bone marrow mesenchymal stem cells. (A)
Representative images of ALP staining and (B) quantification of ALP
activity (n=3; ×5 magnification). (C) Representative images of ARS
staining and (D) quantification of ARS staining (n=3; ×5
magnification). The mRNA expression levels of (E) Col1, (F)
Osterix, (G) Runx2 and (H) Alp (n=3). (I)
Representative western blot images, and semi-quantification of the
protein expression levels of (J) Runx2 and (K) Osterix (n=3). Data
are presented as the mean ± standard deviation. *P<0.05,
**P<0.01, ***P<0.001 and ****P<0.0001. OD, optical
density; Hyp, Hyperoside; Col1, type-I collagen; Runx2,
runt-related transcription factor 2; ARS, Alizarin red S;
ALP/Alp, alkaline phosphatase.

Figure 6

Hyp initiates activation of the
PI3K/AKT signaling pathway. (A) Representative western blotting
images, and semi-quantification of the protein expression levels of
(B) p-PI3K/PI3K and (C) p-AKT/AKT (n=3). (D) Representative
immunohistochemistry images of p-PI3K and p-AKT expression in
femoral sections. Semi-quantification of (E) p-PI3K- and (F)
p-AKT-positive cells observed in (D) (n=5; ×20 magnification). Data
are presented as the mean ± standard deviation. *P<0.05,
**P<0.01, ***P<0.001 and ****P<0.0001. Hyp, Hyperoside;
p-, phosphorylated.

Figure 7

Inhibition of the PI3K/AKT pathway
attenuates the antioxidative and osteogenic effects of Hyp. (A)
Representative fluorescence images of DCFH-DA staining and (B)
quantification (n=3; ×10 magnification). (C) Representative images
of ALP staining and (D) quantification of ALP activity (n=3; ×5
magnification). (E) Representative images of ARS staining and (F)
quantification of ARS absorbance levels (n=3; ×5 magnification).
(G) Representative western blotting images, and semi-quantification
of the protein expression levels of (H) Runx2, (I) Osterix, (J)
p-PI3K/PI3K and (K) p-AKT/AKT (n=3). Data are presented as the mean
± standard deviation. *P<0.05, **P<0.01, ***P<0.001 and
****P<0.0001. Hyp, Hyperoside; ALP, alkaline phosphatase; ARS,
Alizarin red S; Runx2, runt-related transcription factor 2; p-,
phosphorylated.
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Spandidos Publications style
Li L, Deng H, Zhuan S, Wu B and Xiao D: Hyperoside prevents osteoporosis by activating the PI3K/AKT signaling pathway to inhibit oxidative stress and promote osteogenesis. Mol Med Rep 33: 57, 2026.
APA
Li, L., Deng, H., Zhuan, S., Wu, B., & Xiao, D. (2026). Hyperoside prevents osteoporosis by activating the PI3K/AKT signaling pathway to inhibit oxidative stress and promote osteogenesis. Molecular Medicine Reports, 33, 57. https://doi.org/10.3892/mmr.2025.13767
MLA
Li, L., Deng, H., Zhuan, S., Wu, B., Xiao, D."Hyperoside prevents osteoporosis by activating the PI3K/AKT signaling pathway to inhibit oxidative stress and promote osteogenesis". Molecular Medicine Reports 33.2 (2026): 57.
Chicago
Li, L., Deng, H., Zhuan, S., Wu, B., Xiao, D."Hyperoside prevents osteoporosis by activating the PI3K/AKT signaling pathway to inhibit oxidative stress and promote osteogenesis". Molecular Medicine Reports 33, no. 2 (2026): 57. https://doi.org/10.3892/mmr.2025.13767
Copy and paste a formatted citation
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Spandidos Publications style
Li L, Deng H, Zhuan S, Wu B and Xiao D: Hyperoside prevents osteoporosis by activating the PI3K/AKT signaling pathway to inhibit oxidative stress and promote osteogenesis. Mol Med Rep 33: 57, 2026.
APA
Li, L., Deng, H., Zhuan, S., Wu, B., & Xiao, D. (2026). Hyperoside prevents osteoporosis by activating the PI3K/AKT signaling pathway to inhibit oxidative stress and promote osteogenesis. Molecular Medicine Reports, 33, 57. https://doi.org/10.3892/mmr.2025.13767
MLA
Li, L., Deng, H., Zhuan, S., Wu, B., Xiao, D."Hyperoside prevents osteoporosis by activating the PI3K/AKT signaling pathway to inhibit oxidative stress and promote osteogenesis". Molecular Medicine Reports 33.2 (2026): 57.
Chicago
Li, L., Deng, H., Zhuan, S., Wu, B., Xiao, D."Hyperoside prevents osteoporosis by activating the PI3K/AKT signaling pathway to inhibit oxidative stress and promote osteogenesis". Molecular Medicine Reports 33, no. 2 (2026): 57. https://doi.org/10.3892/mmr.2025.13767
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