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Scoparone suppresses mitophagy-mediated NLRP3 inflammasome activation in inflammatory diseases

Acta Pharmacologica Sinica volume 44, pages 1238–1251 (2023)Cite this article

Abstract

Recent evidence shows that targeting NLRP3 inflammasome activation is an important means to treat inflammasome-driven diseases. Scoparone, a natural compound isolated from the Chinese herb Artemisia capillaris Thunb, has anti-inflammatory activity. In this study we investigated the effect of scoparone on NLRP3 inflammasome activation in inflammatory diseases. In LPS-primed, ATP or nigericin-stimulated mouse macrophage J774A.1 cells and bone marrow-derived macrophages (BMDMs), pretreatment with scoparone (50 μM) markedly restrained canonical and noncanonical NLRP3 inflammasome activation, evidenced by suppressed caspase-1 cleavage, GSDMD-mediated pyroptosis, mature IL-1β secretion and the formation of ASC specks. We then conducted a transcriptome analysis in scoparone-pretreated BMDMs, and found that the differentially expressed genes were significantly enriched in mitochondrial reactive oxygen species (ROS) metabolic process, mitochondrial translation and assembly process, as well as in inflammatory response. We demonstrated in J774A.1 cells and BMDMs that scoparone promoted mitophagy, a well-characterized mechanism to control mitochondrial quality and reduce ROS production and subsequent NLRP3 inflammasome activation. Mitophagy blockade by 3-methyladenine (3-MA, 5 mM) reversed the protective effects of scoparone on mitochondrial damage and inflammation in the murine macrophages. Moreover, administration of scoparone (50 mg/kg) exerted significant preventive effects via inhibition of NLRP3 activation in mouse models of bacterial enteritis and septic shock. Collectively, scoparone displays potent anti-inflammatory effects via blocking NLRP3 inflammasome activation through enhancing mitophagy, highlighting a potential action mechanism in treating inflammasome-related diseases for further clinical investigation.

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Fig. 1: Scoparone blocks NLRP3 inflammasome activation in J774A.1 cells and BMDMs.
Fig. 2: Scoparone suppresses NLRP3 inflammasome-mediated pyroptosis in macrophages.
Fig. 3: Scoparone affects mitochondrial signals in response to NLRP3 inflammasome activation.
Fig. 4: Scoparone promotes mitophagy to alleviate mitochondrial damage upon NLRP3 inflammasome stimulation.
Fig. 5: Inhibiting autophagy reverses the protective effects of scoparone against mitochondrial damage and inflammation in murine macrophages.
Fig. 6: Scoparone mitigates E. coli-induced septic shock in vivo.
Fig. 7: Scoparone alleviates enteritis caused by C. rodentium infections in vivo.
Fig. 8: Mechanism of scoparone regulating NLRP3 inflammasome pathway.

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Acknowledgements

This work was supported by the National Natural Science Foundation (NNSF) of China (Nos. 91942301, 31900661 and 82001663), Young Elite Scientists Sponsorship Program by China Association for Science and Technology (No. 2020-QNRC1-03), National Key Research and Development Project (No. 2019YFC1710104), and Joint Fund of Beijing University of Traditional Chinese Medicine and USANA. We thank Dr. Chen Dong for the C. rodentium. We are indebted to all individuals who participated in or helped with this research project.

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  1. These authors contributed equally: Wan-di Feng, Yao Wang, Tong Luo

Authors and Affiliations

  1. Beijing Research Institute of Chinese Medicine, Beijing University of Chinese Medicine, Beijing, 100029, China

    Wan-di Feng, Ting Wang & An-long Xu

  2. Department of Immunology, School of Life Science, Beijing University of Chinese Medicine, Beijing, 100029, China

    Yao Wang, Tong Luo, Cui-qin Cheng, Hao-jia Wang, Xue-jiao Wang, Yi-ying Li, Guang-rui Huang & An-long Xu

  3. National Key Laboratory of Efficacy and Mechanism on Chinese Medicine for Metabolic Diseases, Beijing University of Chinese Medicine, Beijing, 100029, China

    Yao Wang, Ting Wang & An-long Xu

  4. Department of Pharmacology, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, 100029, China

    Xin Jia, Mei-qi Zhang, Qi-qi Li & Jin-yong Wang

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Contributions

ALX conceived the study; WDF and TL designed and performed most of the experiments and collected results. WDF analyzed, interpreted results and wrote the manuscript. YW and XJ designed research and instructed experiment. YW helped to analyze and interprete results, and revised the manuscript. CQC participated in H&E performance. HJW contributed to RNA-seq analysis. MQZ instructed article figure drawing. QQL, XJW, YYL and JYW helped to establish the animal model. GRH and TW oversaw a portion of the work and provided advice. ALX provided funding for the project, and supervised the project and revised/approved the manuscript.

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Correspondence to An-long Xu.

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Feng, Wd., Wang, Y., Luo, T. et al. Scoparone suppresses mitophagy-mediated NLRP3 inflammasome activation in inflammatory diseases. Acta Pharmacol Sin 44, 1238–1251 (2023). https://doi.org/10.1038/s41401-022-01028-9

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