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Nucleolin Mediates LPS-induced Expression of Inflammatory Mediators and Activation of Signaling Pathways

Current Medical Science volume 40, pages 646–653 (2020)Cite this article

Summary

In this study, we investigated the effects of nucleolin on lipopolysaccharide (LPS)-induced activation of MAPK and NF-KappaB (NF-κB) signaling pathways and secretion of TNF-α, IL-1β and HMGB1 in THP-1 monocytes. Immunofluorescence assay and Western blotting were used to identify the nucleolin expression in cell membrane, cytoplasm and nucleus of THP-1 monocytes. Inactivation of nucleolin was induced by neutralizing antibody against nucleolin. THP-1 monocytes were pretreated with anti-nucleolin antibody for 1 h prior to LPS challenge. The irrelevant IgG group was used as control. Secretion of inflammatory mediators (TNF-α, IL-1β and HMGB1) and activation of MAPK and NF-κB/I-κB signaling pathways were examined to assess the effects of nucleolin on LPS-mediated inflammatory response. Nucleolin existed in cell membrane, cytoplasm and nucleus of THP-1 monocytes. Pretreatment of anti-nucleolin antibody significantly inhibited the LPS-induced secretion of TNF-α, IL-1β and HMGB1. P38, JNK, ERK and NF-κB subunit p65 inhibitors could significantly inhibit the secretion of IL-1β, TNF-α and HMGB1 induced by LPS. Moreover, the phosphorylation of p38, JNK, ERK and p65 (or nuclear translocation of p65) was significantly increased after LPS challenge. In contrast, pretreatment of anti-nucleolin antibody could significantly inhibit the LPS-induced phosphorylation of p38, JNK, ERK and p65 (or nuclear translocation of p65). However, the irrelevant IgG, as a negative control, had no effect on LPS-induced secretion of TNF-α and IL-1β and phosphorylation of p38, JNK, ERK and p65 (or nuclear translocation of p65). We demonstrated that nucleolin mediated the LPS-induced activation of MAPK and NF-κB signaling pathways, and regulated the secretion of inflammatory mediators (TNF-α, IL-1β and HMGB1).

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Affiliations

  1. Department of Cardiology, the First Hospital of Changsha, Changsha, 410008, China

    Li Fang, Qing Huang, Feng Cheng, Fang Huang & Wei-wei Liu

  2. Department of Pathophysiology, Xiangya School of Medicine, Central South University, Changsha, 410008, China

    Kang-kai Wang

Authors
  1. Li Fang
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  2. Kang-kai Wang
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  3. Qing Huang
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  4. Feng Cheng
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  5. Fang Huang
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  6. Wei-wei Liu
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Correspondence to Li Fang.

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All authors declare that there is no conflict of interest and financial interest.

Additional information

This work was supported by grants from Bureau of Science and Technology of Changsha, China (No. Kq1701007) and Hunan Natural Science Foundation, China (No. 2018JJ6127).

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Fang, L., Wang, Kk., Huang, Q. et al. Nucleolin Mediates LPS-induced Expression of Inflammatory Mediators and Activation of Signaling Pathways. CURR MED SCI 40, 646–653 (2020). https://doi.org/10.1007/s11596-020-2229-6

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  • DOI: https://doi.org/10.1007/s11596-020-2229-6

Key words

  • nucleolin
  • THP-1 monocytes
  • lipopolysaccharide
  • MAPK
  • NF-κB signaling pathway
  • inflammatory mediators