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Novel HIF-1α Inhibitor AMSP-30m Mitigates the Pathogenic Cellular Behaviors of Hypoxia-Stimulated Fibroblast-Like Synoviocytes and Alleviates Collagen-Induced Arthritis in Rats via Inhibiting Sonic Hedgehog Pathway

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Abstract

Synovial hypoxia-inducible factor 1α (HIF-1α) is a prospective therapeutic target for rheumatoid arthritis (RA). AMSP-30 m, a novel HIF-1α inhibitor, was reported to have notable anti-arthritic effects in rats with adjuvant-induced arthritis. However, its roles in inhibiting the pathogenic behaviors of fibroblast-like synoviocytes (FLS) and the involved mechanisms remain unknown. Here, AMSP-30 m inhibited proliferation and induced apoptosis in hypoxia-induced RA FLS (MH7A cell line), as evidenced by decreased cell viability, reduced Ki67-positive cells, G0/G1 phase arrest, lowered C-myc and Cyclin D1 protein levels, emergence of apoptotic nuclear fragmentation, raised apoptosis rates, and activation of caspase 3. Furthermore, AMSP-30 m prevented hypoxia-induced increases in pro-inflammatory factor production, MMP-2 activity, migration index, migrated/invasive cells, and actin cytoskeletal rearrangement. In vivo, AMSP-30 m alleviated the severity of rat collagen-induced arthritis (CIA). Mechanically, AMSP-30 m reduced HIF-1α expression and blocked sonic hedgehog (Shh) pathway activation in hypoxia-induced MH7A cells and CIA rat synovium, as shown by declines in pathway-related proteins (Shh, Smo, and Gli-1). Particularly, the combination of Shh pathway inhibitor cyclopamine enhanced AMSP-30 m’s inhibitory effects on the pathogenic behaviors of hypoxia-stimulated MH7A cells, whereas the combination of Shh pathway activator SAG canceled AMSP-30 m’s therapeutic effects in vitro and in CIA rats, implying a close involvement of Shh pathway inhibition in its anti-arthritic effects. We likewise confirmed AMSP-30 m’s anti-proliferative role in hypoxia-induced primary CIA FLS. Totally, AMSP-30 m suppressed hypoxia-induced proliferation, inflammation, migration, and invasion of MH7A cells and ameliorated the severity of rat CIA via inhibiting Shh signaling.

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Availability of Data and Materials

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

This work was supported by the National Natural Science Foundation of China (81972040), Anhui Provincial Natural Science Foundation (2108085MH321), The University Synergy Innovation Program of Anhui Province (GXXT-2022–063), Scientific Research Level Promotion Program of Anhui Medical University (2022xkjT008), Open Project Program of Inflammation and Immune Mediated Diseases Laboratory of Anhui Province (IMMDL202001), Research Fund of Anhui Institute of Translational Medicine (2022zhyx-C14), and Undergraduate Innovation Training Program of Anhui Medical University (2021-ZQKY-156, 2022-ZQKY-134).

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Authors and Affiliations

  1. Inflammation and Immune Mediated Diseases Laboratory of Anhui Province, School of Pharmacy, Anhui Medical University, 81 Meishan Road, Hefei, 230032, Anhui Province, People’s Republic of China

    Li Cai, Bo Meng, Fei Jiang, Wen-hao Shu, Xiao-hua Wang, Meng-qing Wang, Xiang Ran & Rong Li

  2. Department of Pathology, School of Basic Medicine, Anhui Medical University, Hefei, 230032, Anhui Province, People’s Republic of China

    Li Cai

  3. The First Clinical Medical College, Anhui Medical University, Hefei, 230032, Anhui Province, People’s Republic of China

    Xin-jie Wu & Yu-chen Yang

  4. The Second Clinical Medical College, Anhui Medical University, Hefei, 230032, Anhui Province, People’s Republic of China

    Ming-wang Hu

  5. Institute of Health and Medicine, Hefei Comprehensive National Science Center, Hefei, 230026, Anhui Province, People’s Republic of China

    Rong Li

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  1. Li Cai
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Contributions

Li Cai: investigation and writing—original draft. Bo Meng, Fei Jiang, Meng-qing Wang, Xin-jie Wu, and Ming-wang Hu: investigation. Wen-hao Shu, Xiao-hua Wang, and Yu-chen Yang: data curation and formal analysis. Xiang Ran: conceptualization, methodology, and visualization. Rong Li: conceptualization, methodology, supervision, writing—review and editing, and funding acquisition. All authors read and approved the manuscript.

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Correspondence to Xiang Ran or Rong Li.

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This study was approved by the Ethical Committee on Animal Research of Anhui Medical University (approval number: LLSC20220564, approval date: July 26, 2022).

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Li Cai, Bo Meng, and Fei Jiang contribute equally to this work

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Cai, L., Meng, B., Jiang, F. et al. Novel HIF-1α Inhibitor AMSP-30m Mitigates the Pathogenic Cellular Behaviors of Hypoxia-Stimulated Fibroblast-Like Synoviocytes and Alleviates Collagen-Induced Arthritis in Rats via Inhibiting Sonic Hedgehog Pathway. Inflammation 46, 2289–2305 (2023). https://doi.org/10.1007/s10753-023-01878-3

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