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Neferine Inhibits Expression of Inflammatory Mediators and Matrix Degrading Enzymes in IL-1β-Treated Rat Chondrocytes via Suppressing MAPK and NF-κB Signaling Pathways

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Abstract

Osteoarthritis (OA), in which inflammation plays a crucial role, is the most common joint disease characterized by cartilage degradation. Neferine (Nef), a dibenzyl isoquinoline alkaloid, has shown its anti-inflammatory effects on other inflammatory diseases. Therefore, we hypothesized that Nef might also have an anti-inflammatory effect on OA and explored its effect on IL-1β-treated rat chondrocytes. Sprague Dawley (SD) rat chondrocytes were stimulated with IL-1β (10 ng/ml) and Nef (1, 5, and 10 μM) or IL-1β (10 ng/ml) alone for 24 h. Expression of inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2), matrix metalloproteinases (MMPs), and thrombospondin motifs-5 (ADAMTS5) was determined by quantitative real-time PCR and Western blotting. Expression of collagen II and aggrecan was examined by Western blotting, immunofluorescence, and safranin O staining. In addition, activation of MAPK and NF-κB signaling pathway was examined by Western blotting, and p65 nuclear translocation was evaluated by immunofluorescence. Nef reduced expression of inflammatory regulators (iNOS and COX-2) in IL-1β-treated chondrocytes. Expression of IL-1β-induced major catabolic enzymes (MMP3, MMP13, and ADAMTS5) was inhibited by Nef. Meanwhile, downregulation of collagen II and aggrecan expression was also ameliorated. Furthermore, Nef dampened abnormal activation of MAPK and NF-κB signaling pathway triggered by IL-1β. Overall, the results above showed that Nef inhibited IL-1β-induced excess production of inflammatory and catabolic factors in rat chondrocytes via inhibiting the MAPK and NF-κB pathways, suggesting a promising pharmacotherapy for OA.

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Funding

National Natural Science Foundation of China (Grant No. 81772390) and the Fundamental Research Funds for the Central Universities (Grant No. 2017KFYXJJ104) funded this research study.

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  1. Department of Orthopedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095#, Jiefang Avenue, Qiaokou District, Wuhan, 430030, Hubei, China

    Bowei Ni, Xiaojian Huang, Yang Xi, Zekai Mao, Xiangyu Chu, Rui Zhang, Xiaohu Ma & Hongbo You

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Contributions

HY and BN conceived the experiments and wrote the manuscript. BN, XH, and YX carried out the cell experiments. ZM and XC purchased animals and isolated primary rat chondrocytes. RZ and XM analyzed the data and made charts and graphs. HY financed the study.

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Correspondence to Hongbo You.

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Ni, B., Huang, X., Xi, Y. et al. Neferine Inhibits Expression of Inflammatory Mediators and Matrix Degrading Enzymes in IL-1β-Treated Rat Chondrocytes via Suppressing MAPK and NF-κB Signaling Pathways. Inflammation 43, 1209–1221 (2020). https://doi.org/10.1007/s10753-019-01143-6

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