endothelin-1 and Periodontitis

Endothelin 1 (ET-1) has been shown to have a key role in homeostasis and function of endothelium and maybe fundamental in the relationship between coronary heart disease (CHD) and periodontitis. In this trial, we assessed the influence on serum and salivary ET-1 levels of gingival health, CHD, periodontitis, or a combination of periodontitis-CHD. Clinical and periodontal parameters, were collected from periodontitis patients (n = 34), CHD patients (n = 34), periodontitis + CHD patients (n = 34), and from healthy patients (n = 34) together with saliva and serum samples. The median concentrations of salivary and serum ET-1 were significantly higher in the CHD patients [serum: 1.4(1.1-1.6) pg/ml; saliva 1.2 (0.9-1.6) µmol/g, p < 0.01] and in the periodontitis + CHD patients [serum: 1.7 (1.2-21.8) pg/ml; salivary 1.4(1-1.6) µmol/g, p < 0.001] respect to periodontitis and control patients. Through a univariate regression analysis, c-reactive protein (CRP) and CHD (both p < 0.001) and periodontitis (p = 0.029) were statistically correlated with ET-1 in serum. The multivariate regression analysis demonstrated that only CRP was the statistically predictor of ET-1 in serum(p < 0.001). The multivariate regression analysis in saliva demonstrated that, regarding ET-1 levels the only predictor were CRP (p < 0.001) and total cholesterol (p = 0.042). The present study evidenced that subjects with CHD and periodontitis plus CHD had higher serum and salivary levels of ET-1 compared to subjects with periodontitis and healthy controls. Moreover, only CRP remained a major predictor of increased ET-1 concentrations in both serum and saliva.

Topics: Adult; Biomarkers; C-Reactive Protein; Case-Control Studies; Coronary Disease; Endothelin-1; Female; Humans; Inflammation Mediators; Male; Middle Aged; Multivariate Analysis; Periodontitis; Saliva

Periodontitis is a very common oral inflammatory disease that results in the destruction of supporting connective and osseous tissues of the teeth. Although the exact etiology is still unclear, Gram-negative bacteria, especially Porphyromonas gingivalis in subgingival pockets are thought to be one of the major etiologic agents of periodontitis. Endothelin (ET) is a family of three 21-amino acid peptides, ET-1, -2, and -3, that activate G protein-coupled receptors, ETA and ETB. Endothelin is involved in the occurrence and progression of various inflammatory diseases. Previous reports have shown that ET-1 and its receptors, ETA and ETB are expressed in the periodontal tissues and, that ET-1 levels in gingival crevicular fluid are increased in periodontitis patients. Moreover, P. gingivalis infection has been shown to induce the production of ET-1 along with other inflammatory cytokines. Despite these studies, however, the functional significance of endothelin in periodontitis is still largely unknown. In this study, we explored the cellular and molecular mechanisms of ET-1 action in periodontitis using human gingival epithelial cells (HGECs). ET-1 and ETA, but not ETB, were abundantly expressed in HGECs. Stimulation of HGECs with P. gingivalis or P. gingivalis lipopolysaccharide increased the expression of ET-1 and ETA suggesting the activation of the endothelin signaling pathway. Production of inflammatory cytokines, IL-1β, TNFα, and IL-6, was significantly enhanced by exogenous ET-1 treatment, and this effect depended on the mitogen-activated protein kinases via intracellular Ca2+ increase, which resulted from the activation of the phospholipase C/inositol 1,4,5-trisphosphate pathway. The inhibition of the endothelin receptor-mediated signaling pathway with the dual receptor inhibitor, bosentan, partially ameliorated alveolar bone loss and immune cell infiltration. These results suggest that endothelin plays an important role in P. gingivalis-mediated periodontitis. Thus, endothelin antagonism may be a potential therapeutic approach for periodontitis treatment.

Topics: Animals; Calcium; Cytokines; Disease Progression; Endothelin-1; Epithelial Cells; Humans; Inflammation; Male; Mice; Periodontitis; Porphyromonas gingivalis; Signal Transduction

Endothelin-1 (ET-1) is a 21-amino acid peptide with multifunctional regulation. Initial research indicated that ET-1 is related to the inflammatory pathogenesis of periodontitis and involved in the regulation of cytokines, but the mechanisms involved remain unclear. The primary aim of this study is to investigate how ET-1 affects proinflammatory cytokine expression in human periodontal ligament (hPDL) cells.. hPDL cells were obtained from both healthy (H)- and periodontitis (P)-affected periodontal tissues. H-hPDL and P-hPDL cells were treated with ET-1 (1, 10, and 100 nM) for 12, 24, and 48 hours. The untreated cells served as a control. To confirm the specificity of the ET-1 effects, 100 nM of the specific endothelin A (ETA) receptor antagonist BQ123 and 100 nM of the specific ETB receptor antagonist BQ788, as negative control, were used. To examine the signaling pathways and molecular mechanisms involved in ET-1-mediated cytokine expression, H-hPDL and P-hPDL cells were pretreated with specific inhibitors for extracellular signal-regulated kinase (ERK1/2) (PD98059), c-Jun N-terminal kinase (SP600125), and p38 kinase (SB203580) for 1 hour before 100 nM ET-1 stimulation. Tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 messenger RNA (mRNA) and protein levels were evaluated by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay, respectively.. ET-1 dose- and time-dependently induced the production of proinflammatory cytokines TNF-α, IL-1β, and IL-6 by H-hPDL and P-hPDL cells at both mRNA and protein levels. However, ETA and ETB receptor antagonists inhibited the stimulatory effects of ET-1 on inflammatory cytokine expression in H-hPDL and P-hPDL cells. Furthermore, inhibitors of the mitogen-activated protein kinases (MAPKs) significantly reduced ET-1-stimulated TNF-α, IL-1β, and IL-6 expression in H-hPDL and P-hPDL cells.. ET-1 may be involved in the inflammatory process of periodontitis, at least in part, by stimulating proinflammatory cytokine production via the MAPK pathway in hPDL cells.

Topics: Adult; Anthracenes; Calcium-Calmodulin-Dependent Protein Kinases; Cell Culture Techniques; Cells, Cultured; Cytokines; Dose-Response Relationship, Drug; Endothelin B Receptor Antagonists; Endothelin Receptor Antagonists; Endothelin-1; Female; Flavonoids; Humans; Imidazoles; Interleukin-1beta; Interleukin-6; JNK Mitogen-Activated Protein Kinases; Male; MAP Kinase Signaling System; Mitogen-Activated Protein Kinase 1; Oligopeptides; p38 Mitogen-Activated Protein Kinases; Peptides, Cyclic; Periodontal Ligament; Periodontitis; Piperidines; Pyridines; Time Factors; Tumor Necrosis Factor-alpha

The combination of obesity and its associated risk factors, such as insulin resistance and inflammation, results in the development of atherosclerosis. However, the effects of periodontitis on atherosclerosis in an obese body remain unclear. The aim of the study was to investigate the effects of ligature-induced periodontitis in Zucker fatty rats on initiation of atherosclerosis by evaluating aortic insulin resistance. Zucker fatty rats (n=24) were divided into two groups. In the periodontitis group, periodontitis was ligature-induced for 4 weeks, whereas the control group was left unligated. After the 4-week experimental period, descending aorta was used for measuring the levels of lipid deposits, immunohistochemical analysis, and evaluation of gene expression. Levels of serum C-reactive protein (CRP), tumor necrosis factor-alpha (TNF-alpha), and insulin were also measured. Rats in the periodontitis group had significantly enhanced lipid deposits in the aorta, but not in the control group. Expression of suppressor of cytokine signaling 3, vascular cell adhesion molecule 1, reactive oxygen species, nitrotyrosine, and endothelin-1 in the periodontitis group was more intense than that in the control group. Significantly decreased levels of phosphatidylinositol 3-kinase (Pi3k) catalytic beta-polypeptide (Pi3kcb), Pi3kp85, and insulin receptor substrate 1 and 2 were observed in the periodontitis group. Levels of serum CRP and TNF-alpha were significantly increased in the periodontitis group. Under insulin-stimulated conditions, aorta in the periodontitis group altered the Akt phosphorylation. Periodontitis in obesity induced the initial stage of atherosclerosis and disturbed aortic insulin signaling.

Topics: Animals; Aorta; Atherosclerosis; Endothelin-1; Gene Expression Profiling; Insulin Resistance; Ligation; Male; Obesity; Oligonucleotide Array Sequence Analysis; Periodontitis; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Rats; Rats, Zucker; Vascular Cell Adhesion Molecule-1; Vascular Endothelial Growth Factor A

Endothelin-1 is a 21-amino-acid peptide with multifunctional regulation. Initial research indicated that endothelin-1 levels in the gingival crevicular fluid from patients with chronic periodontitis were higher than those in the gingival crevicular fluid from healthy subjects. The aim of the present study was to assess the relationship between the clinical parameters and the concentrations of endothelin-1 within the gingival crevicular fluid from inflamed gingiva and periodontitis sites and, subsequently, after the treatment of periodontitis sites.. A total of 60 subjects were divided into three groups - healthy (group I), gingivitis (group II) and chronic periodontitis (group III) - based on gingival index, pocket probing depth and clinical attachment loss. A fourth group consisted of 20 subjects from group III, 6-8 wk after treatment (i.e. scaling and root planing). Gingival crevicular fluid samples collected from each patient were quantified for endothelin-1 using an enzymatic immunometric assay.. Endothelin-1 was not detected in any sample from any of the study groups.. The results showed that all the gingival crevicular fluid samples were negative for the endothelin-1 molecule. Therefore, endothelin-1 cannot be considered as a potential biomarker of periodontal disease progression.

Topics: Adult; Biomarkers; Case-Control Studies; Chronic Disease; Endothelin-1; Female; Gingival Crevicular Fluid; Gingivitis; Humans; Male; Periodontitis

The purpose of this study was to study the expression of endothelin-1 (ET-1) and its receptors ETA and ETB in normal human gingiva and cyclosporin-induced gingival fibroblasts.. Gingival samples were collected from eight normal healthy individuals, eight patients with periodontitis, and eight patients with cyclosporin A (CsA)-induced gingival overgrowth. Total RNA was extracted from tissue samples, and reverse transcriptase-polymerase chain reaction was performed for ET-1, ETA, and ETB. ET-1 protein was estimated from the tissues by enzyme-linked immunosorbent assay. The expression of ET-1 and its receptors was also examined in gingival fibroblast cells treated with CsA.. ET-1 mRNA expression was significantly higher in patients with CsA-induced gingival overgrowth (P <0.001) than in patients with periodontitis and the controls. ETA mRNA was expressed more than the ETB in all examined samples. In human gingival fibroblasts, ET-1 expression was increased with CsA incorporation compared to controls (P <0.001).. These results suggest that CsA can modulate the expression of ET-1 in gingival fibroblasts and CsA-induced gingival overgrowth.

Topics: Adult; Case-Control Studies; Cells, Cultured; Cyclosporine; Endothelin-1; Female; Fibroblasts; Gingival Overgrowth; Humans; Immunosuppressive Agents; Male; Middle Aged; Periodontitis; Receptor, Endothelin A; Reverse Transcriptase Polymerase Chain Reaction

The present study investigated the presence of ET-1 in gingival crevicular fluid (GCF) from patients with periodontitis, and the expression of endothelins (ETs) and their receptors mRNA in cultured cells from human periodontal tissues.. ET was originally discovered as a potent vasoconstrictive peptide from endothelial cells. It has been reported that ETs are produced by various cells besides endothelial cells. ETs are related to inflammatory and sclerotic lesions, such as arteriolosclerosis and hepatic cirrhosis. Therefore, ETs may be involved in periodontal disease. However, the roles of ETs in development and progression of periodontal disease are not clear.. ET-1 released from the cultured cells was measured by enzyme-linked immunosorbent assay. mRNA expressions for ETs and their receptors were examined by reverse transcription-polymerase chain reaction and Northern blotting analysis.. ET-1 levels in GCF from patients with periodontitis were higher than those from healthy subjects. Human gingival keratinocytes (HGK) expressed mRNA for ETs and their receptors, ET-Ar and ET-Br. ET-1 mRNA expression and ET-1 peptide production from HGK were enhanced by interleukin-1beta and tumor necrosis factor-alpha.. These results suggest that ET-1 plays a significant role in periodontal disease.

Topics: Adult; Cells, Cultured; Endothelin-1; Endothelin-2; Endothelins; Female; Gingiva; Gingival Crevicular Fluid; Humans; Inflammation Mediators; Interleukin-1; Keratinocytes; Male; Middle Aged; Periodontitis; Periodontium; Receptor, Endothelin A; Receptor, Endothelin B; Receptors, Endothelin; Tumor Necrosis Factor-alpha

Adult periodontitis, which is the major cause of adult tooth loss, is commonly characterized by chronic inflammatory disease caused by infection with periodontopathic bacteria such as Porphyromonas gingivalis. Our aims in the present study were to examine the expression of endothelin-1 (ET-1) in cultured HEp-2 epithelial cells after infection with P. gingivalis, and in gingival tissue from adult periodontitis patients. The cell lines were infected with the strains P. gingivalis 33277 and 381 for assessment of bacterial invasion using an antibiotic protection assay, and the expression of ET-1, inflammatory cytokines and cell adhesion molecules was examined by ELISA and reverse transcription-PCR. The expression of ET-1, as well as that of interleukin-1 beta, interleukin-8 and ICAM-1 (intercellular cell adhesion molecule 1), was induced significantly in a time-dependent manner, whereas the expression of MCP-1 (monocyte chemotactic protein-1), RANTES (regulated upon activation, normal T-cell expressed and secreted) and VCAM-1 (vascular cell adhesion molecule 1) was not. Furthermore, in gingival tissues from adult periodontitis patients, we also observed increased expression of ET-1 mRNA compared with tissue from normal healthy donors. These results suggest that infection by periodontopathic bacteria up-regulates the expression of ET-1, together with that of inflammatory cytokines and ICAM-1, in gingival epithelial cells, and that ET-1 expression may be closely involved in the regulation of cytokine responses and cell-cell adhesion in adult periodontitis lesions.

Topics: Adult; Case-Control Studies; Cell Line; Endothelin-1; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Gingiva; Humans; Intercellular Adhesion Molecule-1; Interleukin-1; Interleukin-8; Periodontitis; Porphyromonas gingivalis; Reverse Transcriptase Polymerase Chain Reaction

Adult periodontitis is a complex multifactorial disease whose etiology is not well defined. To investigate whether the genes encoded within the HLA class III region may confer susceptibility to periodontitis, polymorphisms in the ET-1 and TNF-beta genes were analyzed together with the I/D polymorphism of the ACE gene.. We determined allele and genotype frequencies of the NcoI bi-allelic polymorphism of the TNF-beta gene, the I/D (insertion/deletion) polymorphism of the ACE gene, and the TaqI polymorphism of the ET-1 gene in 63 Caucasian patients with adult periodontitis and 95 orally healthy controls.. We found a significant difference in a 3 locus combination of genotypes between patients and controls (P<0.05). In the next analyses, no significant differences were found in allele frequencies of single genes, but we did find a significant difference in the genotype distribution between cases and controls for TNF-beta (P<0.03). Differences were also observed for 2 locus combinations of ACE and TNF-beta genotypes (P<0.03), and the ET-1 and TNF-beta (P<0.05) genes. Evidence of deviation from Hardy-Weinberg equilibrium was observed in the periodontitis group for TNF-beta, with an absence of the B1B1 homozygotes in patients.. This study is of an exploratory nature. Considering the number of significant results, however, at least a part of the observed associations may obviously be real and our findings suggest that interactions of the TNF-beta, ET-1, and ACE genes may be involved in susceptibility to adult periodontitis.

Topics: Adult; Alleles; Case-Control Studies; Chi-Square Distribution; Chromosome Mapping; DNA Transposable Elements; Endothelin-1; Female; Gene Frequency; Genetic Predisposition to Disease; Genotype; Homozygote; Humans; Lymphotoxin-alpha; Male; Middle Aged; Odds Ratio; Peptidyl-Dipeptidase A; Periodontitis; Polymorphism, Genetic; Risk Factors; Sequence Deletion; Statistics as Topic