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Natural compound fraxinellone ameliorates intestinal fibrosis in mice via direct intervention of HSP47-collagen interaction in the epithelium

Acta Pharmacologica Sinica volume 44, pages 2469–2478 (2023)Cite this article

Abstract

Intestinal fibrosis is a common complication of inflammatory bowel disease. There is still a lack of effective drugs for the prevention or treatment of intestinal fibrosis. Heat shock protein 47 (HSP47) plays a key role in the development of intestinal fibrosis. In this study we investigated the therapeutic potential and underlying mechanisms of fraxinellone, a degraded limonoid isolated from the root bark of Dictamnus dasycarpus, in the treatment of intestinal fibrosis. Intestinal fibrosis was induced in mice by dextran sodium sulfate (DSS) treatment. DDS-treated mice were administered fraxinellone (7.5, 15, 30 mg·kg−1·d−1, i.g.) for 45 days. We showed that fraxinellone administration dose-dependently alleviated DSS-induced intestinal impairments, and reduced the production of intestinal fibrosis biomarkers such as α-smooth muscle actin (SMA), collagen I, hydroxyproline, fibronectin and laminin, and cytokines such as TGF-β, TNF-α and IL-β. We then established in vitro intestinal fibrosis cell models in SW480 and HT-29 cells, and demonstrated that treatment with fraxinellone (3, 10, 30 μM) significantly relieved TGF-β-induced fibrosis responses by inhibiting the TGF-β/Smad2/3 signaling pathway. Molecular docking suggested that the fraxinellone might disrupt the interaction between HSP47 and collagen, which was confirmed by coimmunoprecipitation experiments. SPR analysis showed that fraxinellone had a high affinity for HSP47 with a Kd value of 3.542 × 10−5 M. This study provides a new example of HSP47-collagen intervention by a natural compound and has important implications for the clinical treatment of inflammation-induced issue fibrosis.

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Fig. 1: Fraxinellone dose-dependently alleviated DSS-induced pathological impairment in mice.
Fig. 2: Fraxinellone treatment inhibited DSS-induced fibrosis in mice.
Fig. 3: Fraxinellone treatment reduced the levels of fibrosis biomarkers and cytokines.
Fig. 4: TGF-β induced the formation of cell fibrosis models in vitro.
Fig. 5: Fraxinellone relieved cellular fibrosis by inhibiting TGF-β/Smad2/3 signaling.
Fig. 6: Fraxinellone reduced the transcription of cellular fibrosis biomarkers.
Fig. 7: Fraxinellone interfered with the HSP47-collagen complex by directly binding to HSP47.
Fig. 8

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (Nos. 82073910, 82173871, 21937005), the Open Fund of State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, China (Grant No. KF-GN-202101), and Fundamental Research Funds for the Central University (021414380503).

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Authors and Affiliations

  1. State Key Laboratory of Pharmaceutical Biotechnology, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, 210000, China

    Jie Wang, Mei Bai, Cui Zhang, Ning An, Li Wan, Xiao-ning Wang, Rong-hui Du, Yan Shen, Zhi-yao Yuan, Xu-dong Wu, Xue-feng Wu & Qiang Xu

  2. School of Basic Medical Sciences, Nanjing Medical University, Nanjing, 210000, China

    Jie Wang & Li Wan

  3. Department of Periodontology, Nanjing Stomatological Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, 210000, China

    Zhi-yao Yuan

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Contributions

JW, XFW, and QX designed research. JW, MB, CZ, NA, and LW performed research and analyzed data. XNW, RHD, YS, XDW, XFW, ZYY, and QX contributed new reagents. JW and XFW wrote the paper. All of the authors have approved the final manuscript.

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Correspondence to Xu-dong Wu, Xue-feng Wu or Qiang Xu.

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Wang, J., Bai, M., Zhang, C. et al. Natural compound fraxinellone ameliorates intestinal fibrosis in mice via direct intervention of HSP47-collagen interaction in the epithelium. Acta Pharmacol Sin 44, 2469–2478 (2023). https://doi.org/10.1038/s41401-023-01143-1

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