1,25-Dihydroxyvitamin D3 Attenuates the Effects of Lipopolysaccharide by Causing ADAM10-Dependent Ectodomain Shedding of Toll-Like Receptor 4

• Published in: Cellular physiology and biochemistry Vol. 41; no. 5; pp. 2104 - 2116
• Main Authors: Yang, Won Seok, Kim, Jin Ju, Han, Nam Jeong, Lee, Eun Kyoung, Park, Su-Kil
• Format: Journal Article
• Language: English
• Published: Basel, Switzerland Cell Physiol Biochem Press GmbH & Co KG 2017
• Subjects: 1,25-Dihydroxyvitamin D3; A disintegrin and metalloprotease 10; ADAM10 Protein - metabolism; Amyloid Precursor Protein Secretases - metabolism; Aorta - metabolism; Aorta - pathology; Calcitriol - pharmacology; Calcium Signaling - drug effects; Cells, Cultured; Ectodomain shedding; Endothelial Cells - metabolism; Endothelial Cells - pathology; Gene Expression Regulation - drug effects; Humans; Intercellular Adhesion Molecule-1 - biosynthesis; L-type calcium channel; Lipopolysaccharide; Lipopolysaccharides - toxicity; Membrane Proteins - metabolism; Original Paper; Protein Domains; Toll-like receptor 4; Toll-Like Receptor 4 - metabolism; Lipopolysaccharide; L-type calcium channel; 1,25-Dihydroxyvitamin D3; Toll-like receptor 4; A disintegrin and metalloprotease 10; Ectodomain shedding
• ISSN: 1015-8987; 1421-9778
• DOI: 10.1159/000475449

Background/Aims: We investigated how 1,25-dihydroxyvitamin D 3 (1,25D 3 ) inhibits the effects of lipopolysaccharide (LPS) in human aortic endothelial cells. Methods: Cellular signaling was explored by determination of protein abundance with Western blot, measurement of cytosolic Ca 2+ concentration and immunofluorescence staining for a disintegrin and metalloprotease 10 (ADAM10). Results: LPS stimulated the expression of intercellular adhesion molecule 1 (ICAM-1) through toll-like receptor 4 (TLR4) and subsequent activation of p38 mitogen-activated protein kinase (p38 MAPK). Pretreatment with 1,25D 3 attenuated LPS-induced p38 MAPK activation and ICAM-1 expression by causing ectodomain shedding of TLR4. This effect of 1,25D 3 depended on its ability to induce a rapid extracellular Ca 2+ influx through L-type calcium channels because the ectodomain shedding was prevented by the absence of extracellular Ca 2+ or the presence of verapamil. TLR4 ectodomain shedding was also induced by Bay K8644 (L-type calcium channel agonist). Both 1,25D 3 and Bay K8644 caused extracellular Ca 2+ influx-dependent ADAM10 translocation to the cell surface. Depletion of ADAM10 by siRNA transfection prevented 1,25D 3 - and Bay K8644-induced ectodomain shedding of TLR4, and abolished the inhibitory effect of 1,25D 3 on LPS-induced ICAM-1 expression. Conclusion: 1,25D 3 causes ectodomain shedding of TLR4 and thereby decreases the responsiveness of cells to LPS. ADAM10, activated by extracellular Ca 2+ influx, was implicated in the ectodomain cleavage of TLR4.