RRC ID 61791
Author Shoji M, Tanabe N, Mitsui N, Tanaka H, Suzuki N, Takeichi O, Sugaya A, Maeno M.
Title Lipopolysaccharide stimulates the production of prostaglandin E2 and the receptor Ep4 in osteoblasts.
Journal Life Sci
Abstract Previous studies have indicated that one of the causes of alveolar bone destruction with periodontitis is lipopolysaccharide (LPS) from the cell wall of gram-negative bacteria in plaque, and that prostaglandin E(2) (PGE(2)) is one of the bone resorption factors that stimulate osteoclast formation through an intercellular interaction between osteoblasts and osteoclast precursors. The present study was undertaken to determine the effect of LPS on cell growth, alkaline phosphatase (ALPase) activity, the production of PGE(2), and the expression of receptors by PGE(2), cyclooxygenase (COX)-1, and COX-2, using human osteosarcoma cell line Saos-2 as osteoblasts. The cells were cultured with 0, 1, or 10 microg mL(-1) of LPS for up to 14 days. The production of PGE(2) and the gene expression of COX-1, COX-2, and PGE(2) receptors, including Ep1, Ep2, Ep3, and Ep4, were determined using enzyme-linked immunosorbent assay (ELISA) and real-time reverse transcription-polymerase chain reaction (real-time RT-PCR), respectively. With the addition of LPS, cell growth and ALPase activity decreased by day 5 of the culture, while PGE(2) production increased in a dose-dependent manner throughout the entire 14-day culture period. LPS-reduced ALP activity and LPS-induced PGE(2) production returned to the control level by the addition simultaneously with indomethacin. The expression of COX-1, Ep1, Ep2, and Ep3 receptors decreased on day 14 of the culture, whereas the expression of COX-2 and Ep4 receptors increased significantly with the addition of LPS. These results suggest that LPS promotes PGE(2) production by increasing the expression of COX-2, and that LPS promotes the production of Ep4 receptors in osteoblasts. These results also indicate that LPS-induced PGE(2) may combine with osteoblast Ep4 receptors in autocrine or paracrine modes, and may promote the formation of osteoclasts.
Volume 78(17)
Pages 2012-8
Published 2006-3-20
DOI 10.1016/j.lfs.2005.09.019
PII S0024-3205(05)01044-1
PMID 16289620
MeSH Alkaline Phosphatase / metabolism Bone Neoplasms Cell Line, Tumor Cell Proliferation / drug effects Cyclooxygenase 1 / genetics Cyclooxygenase 1 / metabolism Cyclooxygenase 2 / genetics Cyclooxygenase 2 / metabolism Dinoprostone / metabolism* Dose-Response Relationship, Drug Enzyme-Linked Immunosorbent Assay Gene Expression / drug effects Humans Indomethacin / pharmacology Lipopolysaccharides / pharmacology* Macrophage Colony-Stimulating Factor / genetics Macrophage Colony-Stimulating Factor / metabolism Membrane Proteins / genetics Membrane Proteins / metabolism Osteoblasts / drug effects* Osteoblasts / metabolism Osteoblasts / pathology Osteosarcoma RNA, Messenger / analysis Receptors, Prostaglandin E / genetics Receptors, Prostaglandin E / metabolism* Receptors, Prostaglandin E, EP4 Subtype Reverse Transcriptase Polymerase Chain Reaction
IF 3.647
Human and Animal Cells Saos-2(RCB0428)