| Recombinant human KAI1/CD82 attenuates M1 macrophage polarization on LPS-stimulated RAW264.7 cells via blocking TLR4/JNK/NF-リB signal pathway |
Hyesook Lee  1,# (Research Professor), Jung-Hwa Han 1,2,# (Post-Doc), Kangbin An1,2 (Graduate student), Yun Jeong Kang1 (Research worker), Hyun Hwangbo 3 (Post-Doc), Ji Hye Heo4 (Professor), Byung Hyun Choi5 (Professor), Jae-Joon Kim6 (Professor), Seo Rin Kim7,# (Professor), Soo Yong Lee8,# (Rofessor), Jin Hur 1,2,9,* (Professor) |
1Department of Convergence Medicine and 2PNU GRAND Convergence Medical Science Education Research Center, Pusan National University School of Medicine,
3Medical Oncology and Hematolog, Department of Internal Medicine, Pusan National University Yangsan Hospital,
4Department of Biomedical Laboratory Science, Donggang University,
5Division of Hepato-Biliary-Pancreatic Surgery and Transplantation, Pusan National University Yangsan Hospital, Pusan National University School of Medicine,
6Medical Oncology and Hematology Department of Internal Medicine and 7Department of Nephrology and Research Institute for Convergence of Biomedical Science and Technology and 8Division of Cardiology, Department of Internal Medicine and Research Institute for Convergence of Bi and 9Research Institute for Convergence of Biomedical Science and Technology, Pusan National University Yangsan Hospital |
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Abstract
KAI1/CD82, a membrane tetraspanin protein, prevents against to various cancers and retinal disorders through its anti-angiogenic and anti-metastatic capacity, yet little is known about its anti-inflammatory effect and its molecular mechanism. Therefore, the aim of the present study was to investigate the effect of a recombinant protein for the large extracellular domain of human KAI1 (Gly 111-Leu 228, rhKAI1) on 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 the expression of classically macrophages (M1) phenotype-related surface markers F4/80+CD86+ in LPS-stimulated BMDM and RAW264.7 cells. In addition, LPS markedly up-regulated the mRNA expression and release levels of pro-inflammatory cytokines and mediators, such as interleukin (IL)-1モ, IL-6, tumor necrosis factor-メ, cyclooxygenase-2, nitric oxide and prostaglandin E2, whereas this increase was substantially down-regulated by rhKAI1. Furthermore, LPS strongly promoted the expression of NF-リB p65 in the nuclei, and increased the phosphorylation of ERK, JNK, and p38 MAPK. However, the nuclear translocation of NF-リB p65 and the phosphorylation of JNK were greatly reversed in the presence of the rhKAI1. Especially, rhKAI1 markedly suppressed the expression of toll-like receptor (TLR4) and prevented the 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, all these findings suggest that rhKAI1 has an anti-inflammatory effect on LPS-polarized macrophages by interacting with TLR4 and down-regulating the JNK/NF-リB signaling pathway.
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| Abstract, Accepted Manuscript(in press) [Submitted on November 15, 2022, Accepted on March 20, 2023] |
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