Recombinant human KAI1/CD82 attenuates M1 macrophage polarization on LPS-stimulated RAW264.7 cells via blocking TLR4/JNK/NF-κB signal pathway
- BMB Rep. 2023 Jun;56(6):359-364. doi: 10.5483/BMBRep.2022-0189.
- 1. Department of Convergence Medicine, Pusan National University School of Medicine, Yangsan 50612, Korea.
- 2. Department of Convergence Medicine, Pusan National University School of Medicine, Yangsan 50612; PNU GRAND Convergence Medical Science Education Research Center, Pusan National University School of Medicine, Yangsan 50612, Korea.
- 3. Department of Biochemistry, Dong-Eui University College of Korean Medicine, Busan 47227, Korea.
- 4. Department of Biomedical Laboratory Science, Donggang University, Gwangju 61200, Korea.
- 5. 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, Yangsan 50612, Korea.
- 6. Medical Oncology and Hematology, Department of Internal Medicine, Pusan National University Yangsan Hospital, Yangsan 50612, Korea.
- 7. Division of Nephrology, Department of Internal Medicine, Pusan National University School of Medicine, Yangsan 50612, Korea.
- 8. 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, Yangsan 50612, Korea.
- 9. Department of Convergence Medicine, Pusan National University School of Medicine, Yangsan 50612; PNU GRAND Convergence Medical Science Education Research Center, Pusan National University School of Medicine, Yangsan 50612; Research Institute for Convergence of Biomedical Science and Technology, Pusan National University Yangsan Hospital, Yangsan 50612, Korea.
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) phenotyperelated 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. [BMB Reports 2023; 56(6): 359-364].