Expression of Phospholipase D in Periodontitis and Its Role in the Inflammatory and Osteoclastic Response by Nicotine- and Lipopolysaccharide-Stimulated Human Periodontal Ligament Cells
Abstract
The aim of the present study is to investigate the expression of phospholipase D (PLD) 1 and PLD2 in periodontal patients and in human periodontal ligament cells (HPDLCs) exposed to nicotine plus lipopolysaccharide (LPS) from Porphyromonas gingivalis (Toll-like receptor 2 ligand). Furthermore, the effects of PLD isoform inhibition on the inflammatory response and osteoclast differentiation and its mechanisms were determined. Proinflammatory mediators were examined by reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay. To silence the gene expression of the PLD isoforms, cells were transfected with small interfering RNA (siRNA) targeting PLD1 or PLD2. Mouse bone marrow-derived macrophages (BMMs) were used as osteoclast precursor cells for in vitro osteoclastogenesis. Western blot analysis and immunofluorescence were used to assess signaling pathways. Chronic smokers with periodontitis exhibited significantly higher PLD1 and PLD2 messenger RNA (mRNA) expression than non-smokers with periodontitis and healthy controls. Nicotine and LPS upregulated PLD1 and PLD2 mRNA expression in a dose-dependent manner in HPDLCs. Pharmacologic and siRNA-mediated inhibition of PLD1 and PLD2 attenuated the nicotine...Continue Reading
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