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Protective Effect of Silibinin on Lipopolysaccharide-Induced Endotoxemia by Inhibiting Caspase-11-Dependent Cell Pyroptosis

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Chinese Journal of Integrative Medicine Aims and scope Submit manuscript

Abstract

Objective

To explore the protective effect and the underlying mechanism of silibinin (SIB), one of the active compounds from Silybum marianum (L.) Gaertn in endotoxemia.

Methods

Mouse peritoneal macrophage were isolated via intraperitoneally injection of BALB/c mice with thioglycolate medium. Cell viability was assessed using the cell counting kit-8, while cytotoxicity was determined through lactate dehydrogenase cytotoxicity assay. The protein expressions of interleukin (IL)-1 α, IL-1 β, and IL-18 were determined by enzyme-linked immunosorbent assay. Intracellular lipopolysaccharide (LPS) levels were measured by employing both the limulus amoebocyte lysate assay and flow cytometry. Additionally, proximity ligation assay was employed for the LPS and caspase-11 interaction. Mice were divided into 4 groups: the control, LPS, high-dose-SIB (100 mg/kg), and low-dose-SIB (100 mg/kg) groups (n=8). Zebrafish were divided into 4 groups: the control, LPS, high-dose-SIB (200 εmol/L), and low-dose-SIB (100 εmol/L) groups (n=30 for survival experiment and n=10 for gene expression analysis). The expression of caspase-11, gasdermin D (GSDMD), and N-GSDMD was determined by Western blot and the expressions of caspy2, gsdmeb, and IL-1 β were detected using quantitative real-time PCR. Histopathological observation was performed through hematoxylineosin staining, and protein levels in bronchoalveolar lavage fluid were quantified using the bicinchoninicacid protein assay.

Results

SIB noticeably decreased caspase-11 and GSDMD-mediated pyroptosis and suppressed the secretion of IL-1 α, IL-1 β, and IL-18 induced by LPS (P<0.05). Moreover, SIB inhibited the translocation of LPS into the cytoplasm and the binding of caspase-11 and intracellular LPS (P<0.05). SIB also attenuated the expression of caspase-11 and N-terminal fragments of GSDMD, inhibited the relative cytokines, prolonged the survival time, and up-regulated the survival rate in the endotoxemia models (P<0.05).

Conclusions

SIB can inhibit pyroptosis in the LPS-mediated endotoxemia model, at least in part, by inhibiting the caspase-11-mediated cleavage of GSDMD. Additionally, SIB inhibits the interaction of LPS and caspase-11 and inhibits the LPS-mediated up-regulation of caspase-11 expression, which relieves caspase-11-dependent cell pyroptosis and consequently attenuates LPS-mediated lethality.

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Data Availability

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

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Author information

Authors and Affiliations

  1. Third Level Research Laboratory of State Administration of Traditional Chinese Medicine, School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, 510515, China

    Jin-ying Ou, Shan-hong Liu, Dong-kai Tang, Ling-zhu Shi, Li-jun Yan, Jing-yan Huang, Li-fang Zou, Jing-yu Quan, Yu-yao Chen, Lin-zhong Yu & Zi-bin Lu

  2. Guangdong Provincial Key Laboratory of Chinese Medicine Pharmaceutics, Guangzhou, 510515, China

    Jin-ying Ou, Shan-hong Liu, Dong-kai Tang, Ling-zhu Shi, Li-jun Yan, Jing-yan Huang, Li-fang Zou, Jing-yu Quan, Yu-yao Chen, Lin-zhong Yu & Zi-bin Lu

  3. School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, 510515, China

    Yan-ting You

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  1. Jin-ying Ou
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Contributions

Ou JY and Liu SH designed and conducted the experiment. Paper drafting and a portion of the in vivo experiment were carried out by Ou JY, Liu SH, Tang DK, Shi LZ, and Chen YY. Yan LJ, Huang JY, and You YT were responsible for data curation and visualization. Zou LF, You YT, Quan JY, and Yu LZ handled data curation, formal analysis, and methodology. Lu ZB provided supervision, research guidance, and reviewed the draft. All authors read and approved the final manuscript for publication.

Corresponding author

Correspondence to Zi-bin Lu.

Ethics declarations

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Additional information

Supported by the Guangdong Basic and Applied Basic Research Foundation (No. 2023A1515011106), Science and Technology Program of Guangzhou (No. 2023A04J1826)

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Ou, Jy., Liu, Sh., Tang, Dk. et al. Protective Effect of Silibinin on Lipopolysaccharide-Induced Endotoxemia by Inhibiting Caspase-11-Dependent Cell Pyroptosis. Chin. J. Integr. Med. (2024). https://doi.org/10.1007/s11655-024-3656-1

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  • DOI: https://doi.org/10.1007/s11655-024-3656-1

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