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Calycosin prevents IL-1β-induced articular chondrocyte damage in osteoarthritis through regulating the PI3K/AKT/FoxO1 pathway

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Abstract

Osteoarthritis (OA) is a joint disorder that is associated with chondrocyte damage under inflammatory environment. Calycosin is an astragalus extract with potential anti-inflammatory and anti-tumor activities. The purpose of this research is to explore the activity and mechanism of calycosin in interleukin-1beta (IL-1β)-induced chondrocyte injury. In the present study, the targets of calycosin and OA were analyzed according to HERB, DisGeNet, String, GO terms, and KEGG pathway enrichment assays. Human primary chondrocytes were treated with calycosin, and stimulated with IL-1β. Cell viability was detected by CCK-8 assay. Cell apoptosis was investigated by flow cytometry, and caspase-3 activity analyses. Inflammation was analyzed according to inflammatory cytokines levels by enzyme-linked immunosorbent assay (ELISA). The proteins associated with extracellular matrix (ECM) degradation and phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/forkhead box O1 (FoxO1) signaling pathways were measured using Western blotting. The results showed that total of 25 overlapping targets of calycosin against OA were predicted. These targets might drive the FoxO pathway. Calycosin alone induced little cytotoxicity to chondrocytes, and it alleviated IL-1β-induced viability inhibition, cell apoptosis, inflammatory cytokine secretion, and ECM degradation in chondrocytes. Calycosin repressed IL-1β-induced activation of the PI3K/AKT/FoxO1 signaling. Activation of the PI3K/AKT/FoxO1 signaling mitigated the suppressive effect of calycosin on chondrocyte apoptosis, inflammation, and ECM degradation induced by IL-1β. As a conclusion, calycosin prevents IL-1β-induced chondrocyte apoptosis, inflammation, and ECM degradation through inactivating the PI3K/AKT/FoxO1 pathway.

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Data availability

The data and material presented in this manuscript are available from the corresponding author on reasonable request.

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Acknowledgements

I would like to thank Ms. Xiaoyu Pan, Jianhong Wu, Yuanzhou Li, and Na Nie, who has helped me during the writing, design, and experiment performance of this thesis.

Funding

This research was supported by Research Initiation Project of Shaoxing University of Arts and Sciences (No. 20210004) and the National College Students Innovation and Entrepreneurship Training Program in 2020 (No. 202010349051X)

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Authors and Affiliations

  1. School of medicine, Shaoxing University, Zhejiang, 312000, Shaoxing, China

    Xiang Guo & Jianhong Wu

  2. Department of Clinical Medicine, The Medical College of Shaoxing University, Shaoxing, 312000, Zhejiang, China

    Xiaoyu Pan

  3. Shaoxing Geke Biological Technology Co. Ltd, Shaoxing, 312000, Zhejiang, China

    Yuanzhou Li

  4. Trauma Joint Surgery, the Third Affiliated Hospital of Chongqing Medical University, Chongqing, 404100, China

    Na Nie

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  1. Xiang Guo
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  5. Na Nie
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Contributions

Xiang Guo designed and performed most of the experiments and wrote the manuscript. Xiaoyu Pan, Jianhong Wu, Yuanzhou Li, and Na Nie were involved in the performance of this experiment.

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Correspondence to Xiang Guo.

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Supplementary file1

The influence of FoxO1 silencing on influence of calycosin in IL-1β-treated chondrocytes. Chondrocytes were transfected with si-con or si-FoxO1, and then treated with IL-1β and calycosin. (A) Apoptosis was measured by flow cytometry. (B) Relative caspase-3 activity was detected. (C) TNF-α, IL-6 and IL-8 levels were detected by ELISA. (PNG 321 kb)

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Guo, X., Pan, X., Wu, J. et al. Calycosin prevents IL-1β-induced articular chondrocyte damage in osteoarthritis through regulating the PI3K/AKT/FoxO1 pathway. In Vitro Cell.Dev.Biol.-Animal 58, 491–502 (2022). https://doi.org/10.1007/s11626-022-00694-7

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  • DOI: https://doi.org/10.1007/s11626-022-00694-7

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