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Formononetin ameliorates the LPS-induced inflammatory response and apoptosis of neuronal cells via NF-κB/NLRP3 signaling pathway

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Abstract

The objective of this study was to investigate the impact of formononetin on cellular apoptosis and inflammatory responses following spinal cord injury (SCI), as well as the underlying mechanisms involved. In this study, PC12 cells were treated with lipopolysaccharide (LPS) and different concentrations of Formononetin (FT) (50 μM, 100 μM, 200 μM). To confirm the effect of nuclear factor-κB (NF-κB)/NLR family pyrin domain containing 3 (NLRP3) signaling pathways, the cells in the phorbol-12-myristate-13-acetate (PMA) group were treated with 0.1 μmol/L PMA (NF-κB/NLRP3 signaling pathway activators). The lactate dehydrogenase (LDH) concentration and cell viability, proliferating cell nuclear antigen (PCNA) fluorescence intensity, and cell apoptosis were determined using an LDH kit, Cell Counting Kit-8 (CCK-8), immunofluorescence, and flow cytometry assays, respectively. Tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-16 (IL-6) expression levels were detected by quantitative ELISA assay. The expression of proteins related to the NF-κB/NLRP3 signaling pathway was detected by western blotting. Our results showed that LPS increased LDH levels in PC12 cells, suggesting that inflammation caused PC12 cell damage. However, the PC12 cell damage was decreased by methylprednisolone. Formononetin promotes cell survival and proliferation, and prevents apoptosis in a concentration-dependent manner. Formononetin reduced the TNF-α, IL-1β, and IL-6 levels in the LPS-treated model. Moreover, formononetin decreased the levels of p-p65 NF-κB and NLRP3 in PC12 cells. We conclude that formononetin ameliorated the inflammatory response and apoptosis in LPS-induced inflammatory injury in neuronal cells via the NF-κB/NLRP3 signaling pathway.

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Data availability

The data used to support the findings of this study are available from the corresponding author upon request.

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Authors and Affiliations

  1. Department of Orthopedic, Lianyungang Hospital of Traditional Chinese Medicine, Chaoyang Middle Road No. 160, Haizhou District, Lianyungang, 222004, Jiangsu, China

    Zhijing Zhou & Peng Zhang

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  1. Zhijing Zhou
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  2. Peng Zhang
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Contributions

PZ conceived and designed the study. ZZ experimented and drafted the manuscript. ZZ and PZ performed the literature search and data analysis. ZZ and PZ revised the manuscript. All authors reviewed the manuscript.

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Correspondence to Peng Zhang.

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The experimental protocols were approved by the Ethics Committee of the Lianyungang Hospital of Traditional Chinese Medicine. No patient was involved in this study.

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Zhou, Z., Zhang, P. Formononetin ameliorates the LPS-induced inflammatory response and apoptosis of neuronal cells via NF-κB/NLRP3 signaling pathway. Funct Integr Genomics 23, 321 (2023). https://doi.org/10.1007/s10142-023-01247-1

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  • DOI: https://doi.org/10.1007/s10142-023-01247-1

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