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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Recombinant human KAI1/CD82 attenuates M1 macrophage polarization on LPS-stimulated RAW264.7 cells via blocking TLR4/JNK/NF-κB signal pathway

  • Hyesook Lee (Department of Convergence Medicine, Pusan National University School of Medicine) ;
  • Jung-Hwa Han (Department of Convergence Medicine, Pusan National University School of Medicine) ;
  • Kangbin An (Department of Convergence Medicine, Pusan National University School of Medicine) ;
  • Yun Jeong Kang (Department of Convergence Medicine, Pusan National University School of Medicine) ;
  • Hyun Hwangbo (Department of Biochemistry, Dong-Eui University College of Korean Medicine) ;
  • Ji Hye Heo (Department of Biomedical Laboratory Science, Donggang University) ;
  • Byung Hyun Choi (Division of Hepato-Biliary-Pancreatic Surgery and Transplantation, Department of Surgery, Research Institute for Convergence of Biomedical Science and Technology, Pusan National University Yangsan Hospital, Pusan National University School of Medicine) ;
  • Jae-Joon Kim (Medical Oncology and Hematology, Department of Internal Medicine, Pusan National University Yangsan Hospital) ;
  • Seo Rin Kim (Division of Nephrology, Department of Internal Medicine, Pusan National University School of Medicine) ;
  • Soo Yong Lee (Division of Cardiology, Department of Internal Medicine, Research Institute for Convergence of Biomedical Science and Technology, Pusan National University Yangsan Hospital, Pusan National University School of Medicine) ;
  • Jin Hur (Department of Convergence Medicine, Pusan National University School of Medicine)
  • Received : 2022.11.15
  • Accepted : 2023.03.20
  • Published : 2023.06.30
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Abstract

KAI1/CD82, a membrane tetraspanin protein, can prevent various cancers and retinal disorders through its anti-angiogenic and anti-metastatic capacity. However, little is known about its anti-inflammatory effect and molecular mechanism. Therefore, the present study aimed to inLPSvestigate effect of a recombinant protein of the large extracellular domain of human KAI1 (Gly 111-Leu 228, rhKAI1) on lipopolysaccharides (LPS)-stimulated RAW264.7 macrophage-like cells and mouse bone marrow-derived macrophages (BMDM) and to identify its underlying mechanism. Our data showed that rhKAI1 suppressed expression levels of classically macrophages (M1) phenotype-related surface markers F4/80+CD86+ in LPS-stimulated BMDM and RAW264.7 cells. In addition, LPS markedly increased mRNA expression and release levels of pro-inflammatory cytokines and mediators such as interleukin (IL)-1β, IL-6, tumor necrosis factor-α, cyclooxygenase-2, nitric oxide and prostaglandin E2, whereas these increases were substantially down-regulated by rhKAI1. Furthermore, LPS strongly increased expression of NF-κB p65 in the nuclei and phosphorylation of ERK, JNK, and p38 MAPK. However, nuclear translocation of NF-κB p65 and phosphorylation of JNK were greatly reversed in the presence of rhKAI1. Especially, rhKAI1 markedly suppressed expression of toll-like receptor (TLR4) and prevented binding of LPS with TLR4 through molecular docking predict analysis. Importantly, Glu 214 of rhKAI1 residue strongly interacted with Lys 360 of TLR4 residue, with a binding distance of 2.9 Å. Taken together, these findings suggest that rhKAI1 has an anti-inflammatory effect on LPS-polarized macrophages by interacting with TLR4 and down-regulating the JNK/NF-κB signaling pathway.

Keywords

Acknowledgement

This research was supported by the Korean Fund for Regenerative Medicine (KFRM) grant funded by the Korea government (the Ministry of Science and ICT, the Ministry of Health & Welfare; grant No. KFRM 21A0502L1-12) and by Research institute for Convergence of biomedical science and technology of Pusan National University Yangsan Hospital. Fig. 4 was created in part with BioRender.com.

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