RESUMO
OBJECTIVE: To evaluate the expression of the cytokines IFN-γ, IL-1ß, IL-4, IL-6, IL-10, IL-21, IL-22, IL-23, IL-25, IL-31, IL-33, IL-17A, IL-17F, sCD40L, and TNF-α in gingival tissue and intestinal mucosa of patients having both periodontitis and inflammatory bowel disease (IBD) and assess how they cluster in both tissues. METHODS: This cross-sectional study selected 28 patients with periodontitis (18 with Crohn's disease and 10 with ulcerative colitis) from the IBD gastroenterology outpatient clinic at the Pedro Ernesto University Hospital. Patients were assessed using questionnaire, medical chart check and periodontal examination. Gingival and intestinal biopsies were collected and homogenized using a cell disruptor. Cytokines expression was evaluated through multiplex technology. Cluster analysis was performed based on cytokinés correlation strength and presented in dendrograms. RESULTS: Crohn's disease and ulcerative colitis patients exhibited no significant difference between them in cytokine levels (p>0.05), so they were analysed together. Significantly higher levels of IL-17A, IL-17F, IL-22, IL-25, IL-33, IL-10, and INF-γ were found in gingival tissues in comparison with intestinal mucosa (p<0.05). In gingival tissue, cytokines formed the following clusters: IL-25/IL-10/IL-33 (r=0.775), IL-22/IL-23/IL-6 (r=0.681) and IL-6/IL-25/IL-33/IL-10 (r=0.660). In intestinal mucosa, the following clusters were formed: IL-6/IL-21/IL-10 (r=0.880), IL-17A/IL-6/IL-21/IL-10 (r=0.826), IL-I7F/IL-33/IL-25 (r=0.813) and IL-23/IL-2/IL-17A/IL-6/IL-21/IL-10 (r=0.785). CONCLUSION: Expression of IL-17A, IL-17F, IL-22, IL-25, IL-33, IL-10, and INF-γ was significantly increased in gingival tissue in comparison with intestinal mucosa of patients with periodontitis and IBD. The cytokine clustering pattern was different in gingival and intestinal tissues.
Assuntos
Citocinas/biossíntese , Gengiva/metabolismo , Doenças Inflamatórias Intestinais/metabolismo , Mucosa Intestinal/metabolismo , Periodontite/metabolismo , Adulto , Idoso , Biópsia , Colite Ulcerativa/imunologia , Colite Ulcerativa/metabolismo , Doença de Crohn/imunologia , Doença de Crohn/metabolismo , Estudos Transversais , Citocinas/metabolismo , Feminino , Gengiva/imunologia , Humanos , Doenças Inflamatórias Intestinais/complicações , Doenças Inflamatórias Intestinais/imunologia , Interferon gama/biossíntese , Interferon gama/imunologia , Interleucinas/biossíntese , Mucosa Intestinal/imunologia , Masculino , Pessoa de Meia-Idade , Periodontite/complicações , Periodontite/imunologia , Fator de Necrose Tumoral alfa/biossíntese , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Several studies have suggested an increase of cardiovascular disease (CVD) risk on periodontitis patients. An enhancement has been demonstrated on both platelet activation and oxidative stress on periodontitis patients, which may contribute for this association. Therefore, the aim of this study was to evaluate the effects of non-surgical periodontal treatment on the l-arginine-nitric oxide (NO)-cyclic guanosine monophosphate (cGMP) pathway and oxidative status in platelets. A total of eight periodontitis patients and eight controls were included in this study. Clinical, laboratory and experimental evaluations were performed on baseline and 90 days after periodontal treatment (except for western blot analysis). The clinical periodontal evaluation included measurements of probing pocket depth (PPD), clinical attachment loss (CAL), % of sites with plaque and % of sites with bleeding on probing. We evaluated: l-[(3)H]arginine influx; nitric oxide synthase (NOS) and arginase enzymes activity and expression; expression of guanylate cyclase and phosphodiesterase-5 enzymes; cGMP levels; platelet aggregation; oxidative status through superoxide dismutase (SOD) and catalase activities, and measurement of reactive oxygen species (ROS) levels and C-reactive protein (CRP) levels. The initial results showed an activation of both l-arginine influx and via system y (+ )L associated with reduced intraplatelet cGMP levels in periodontitis patients and increased systemic levels of CRP. After periodontal treatment, there was a significant reduction of the % of sites with PPD 4-5mm, % of sites with CAL 4-5 mm, and an enhancement in cGMP levels and SOD activity. Moreover, CRP levels were reduced after treatment. Therefore, alterations in the intraplatelet l-arginine-NO-cGMP pathway and oxidant-antioxidant balance associated with a systemic inflammatory response may lead to platelet dysfunction, which may contribute to a higher risk of CVD in periodontitis.
Assuntos
Arginina/metabolismo , Plaquetas/metabolismo , Óxido Nítrico/metabolismo , Estresse Oxidativo/fisiologia , Periodontite/metabolismo , Adulto , Proteína C-Reativa/metabolismo , Doenças Cardiovasculares/complicações , Doenças Cardiovasculares/metabolismo , GMP Cíclico/metabolismo , Humanos , Pessoa de Meia-Idade , Óxido Nítrico Sintase/metabolismo , Periodontite/complicações , Ativação Plaquetária/fisiologia , Agregação Plaquetária/fisiologia , Transdução de Sinais/fisiologiaRESUMO
BACKGROUND: Major depression (MD) is an independent cardiovascular risk factor, but the exact mechanisms are not clear. In this study we have investigated the intraplatelet L-arginine-nitric oxide (NO) pathway and platelet function in depressive patients. METHODS: Nineteen unmedicated patients with MD (34±4years) and 19 control subjects (CS, 34±3years) were included. L-[(3)H]-arginine influx, NO synthase (NOS) activity and intracellular cGMP levels were evaluated in platelets, as well as the expression of eNOS, iNOS, arginase and soluble guanylate cyclase (sGC), platelet aggregation and the systemic amino acid profile in MD patients and CS. RESULTS: L-arginine influx (pmol/10(9)cells/min) in platelets was reduced from 46.2±9.5 to 20.02±2.12 in depression. NOS activity (pmol/10(8) cells) was diminished in MD patients (0.09±0.01) compared to CS (0.17±0.01). Intracellular cGMP levels were also impaired in MD patients associated with hyperaggregability. Moreover, the concentration of plasma L-arginine was reduced by 20% in MD patients. The expression of eNOS, iNOS, arginase II and sGC in platelet lysates was not affected by MD. LIMITATIONS: Small number of patients in the study. CONCLUSIONS: This study has demonstrated an impairment of L-arginine-NO signaling in platelets from MD patients, suggesting a role in platelet activation and cardiovascular events.
Assuntos
Arginina/sangue , Plaquetas/fisiologia , Doenças Cardiovasculares/metabolismo , Transtorno Depressivo Maior/metabolismo , Óxido Nítrico/metabolismo , Agregação Plaquetária/fisiologia , Adulto , Aminoácidos/sangue , Arginina/metabolismo , Plaquetas/química , Plaquetas/metabolismo , Doenças Cardiovasculares/etiologia , Doenças Cardiovasculares/fisiopatologia , GMP Cíclico/metabolismo , Transtorno Depressivo Maior/complicações , Transtorno Depressivo Maior/fisiopatologia , Feminino , Guanilato Ciclase/metabolismo , Humanos , Masculino , Óxido Nítrico/sangue , Óxido Nítrico Sintase/metabolismo , Fatores de RiscoRESUMO
Nitric oxide (NO) production occurs through oxidation of the amino acid L-arginine by NO synthase (NOS). NO inhibits platelet activation by increasing the levels of cyclic guanosine monophosphate (cGMP), thus maintaining vascular homeostasis. Our group previously demonstrated (da Silva et al. 2005) an enhancement of the L-arginine-NO-cGMP pathway in platelets taken from chronic renal failure (CRF) patients on haemodialysis associated with reduced platelet aggregation. We investigate the platelet L-arginine-NO-cGMP pathway, platelet function, and inflammation from patients in CRF on conservative treatment. A total of 42 CRF patients and 42 controls (creatinine clearance = 27 ± 3 vs. 93 ± 1 mL per min per 1.73 m2, respectively) participated in this study. NOS activity and expression and cGMP concentration were measured in platelets. Platelet aggregation induced by collagen or ADP was evaluated and plasma levels of fibrinogen were determined by the Clauss method. A marked increase in basal NOS activity was seen in undialysed CRF patients compared with controls, accompanied by an elevation of fibrinogen plasma levels. There were no differences in expression of NOS and in cGMP levels. In this context, platelet aggregation was not affected. We provide the first evidence of increased intraplatelet NO biosynthesis in undialysed CRF patients, which can be an early marker of future haemostatic abnormalities during dialysis treatment.
Assuntos
Plaquetas/metabolismo , Falência Renal Crônica/sangue , Óxido Nítrico/biossíntese , Óxido Nítrico/sangue , Difosfato de Adenosina/farmacologia , Arginina/sangue , Estudos de Casos e Controles , Colágeno/farmacologia , GMP Cíclico/sangue , Feminino , Fibrinogênio/metabolismo , Humanos , Inflamação/sangue , Falência Renal Crônica/enzimologia , Masculino , Pessoa de Meia-Idade , Óxido Nítrico Sintase Tipo II/biossíntese , Óxido Nítrico Sintase Tipo II/sangue , Óxido Nítrico Sintase Tipo III/biossíntese , Óxido Nítrico Sintase Tipo III/sangue , Agregação Plaquetária/efeitos dos fármacosRESUMO
Estudos publicados nas duas últimas décadas sugerem um aumento do risco de doença cardiovascular (DCV) em pacientes com periodontite, mas os mecanismos fisiopatológicos dessa associação ainda não estão completamente esclarecidos. Uma vez que foi demonstrado aumento da ativação plaquetária e do estresse oxidativo na periodontite, o objetivo do presente estudo foi investigar a via L-arginina-óxido nítrico (NO)- guanosina monofosfato cíclica (GMPc) e parâmetros de estresse oxidativo em plaquetas de pacientes com periodontite, bem como avaliar o efeito do tratamento periodontal não-cirúrgico nessas variáveis. Um total de 10 pacientes sem periodontite (periodontalmente saudáveis ou com gengivite) e 10 pacientes com periodontite participaram do estudo. A avaliação clínica, laboratorial e experimental foi realizada no início do estudo e 90 dias após realização da terapia periodontal básica (grupo periodontite). A avaliação clínica periodontal incluiu registros de: profundidade de bolsa à sondagem (PBS), nível de inserção (NIC), percentual de placa e percentual de sangramento à sondagem. Os seguintes experimentos foram realizados: influxo de L-arginina; atividade e expressão das enzimas óxido nítrico sintase e da arginase; expressão das enzimas guanilato ciclase solúvel e fosfodiesterase 5; determinação dos níveis intraplaquetários de GMPc; agregação plaquetária; avaliação do estresse oxidativo (atividade oxidante total, atividade das enzimas antioxidantes catalase e da superóxido dismutase - SOD); medição dos níveis de proteína C reativa (CRP) e de fibrinogênio. Os resultados obtidos no início do estudo demonstraram ativação do influxo de L-arginina em plaquetas via sistema y+L nos pacientes com periodontite, bem como concentrações intraplaquetárias de GMPc diminuídas e aumento sistêmico da CRP. Após o tratamento periodontal, observou-se redução do percentual de sítios com PBS ≥ 6 mm, NIC 4-5 mm e NIC ≥ 6 mm, aumento nos níveis de GMPc, para níveis ...
Studies published over the last two decades have suggested an increase of cardiovascular disease (CVD) risk on periodontitis patients, but the physiopathological mechanisms involved in this association are not yet clear. Since it has been demonstrated an enhancement on both platelet activation and oxidative stress on periodontitis patients, the aim of this study was to investigate the L-arginine-nitric oxide (NO)-cyclic guanosine monophosphate (cGMP) pathway on platelets from periodontitis patients, and the effect of non-surgical periodontal treatment in these variables. A total of 10 patients without periodontitis (periodontal healthy controls or gingivitis patients) and 10 periodontitis patients were included in this study. The clinical, laboratorial, and experimental evaluations were performed at the beginning of the study and 90 days after the basic periodontal therapy (periodontitis group). The clinical periodontal evaluation included the measurements of probing pocket depth (PPD), clinical attachment level (CAL), plaque percentage, and percentage of bleeding on probing. The following experiments were performed: L-arginine influx; nitric oxide synthase and arginase enzymes activity and expression; expression of guanylate cyclase and phosphodiesterase-5 enzymes; measurement of intraplatelet cGMP levels; platelet aggregation; oxidative stress evaluation (total oxidant activity and activity of both antioxidant enzymes catalase and superoxide dismutase SOD); measurement of C reactive protein (CRP) and fibrinogen. The initial results demonstrated an activation of L-arginine influx in platelets from periodontitis patients via y+L system, reduced intraplatelet cGMP levels and increased CRP. After periodontal treatment, it was observed reduction on percentage of sites with PPD ≥ 6 mm, CAL 4-5 mm and CAL ≥ 6 mm, enhancement on cGMP levels, to levels comparables to patients without periodontitis, accompanied by a higher activity of both antioxidant ...(AU)
Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Arginina/metabolismo , GMP Cíclico/metabolismo , Estresse Oxidativo , Óxido Nítrico/metabolismo , Periodontite/terapia , Plaquetas/enzimologia , Plaquetas/metabolismo , Proteína C-Reativa , Doenças Cardiovasculares/etiologia , Fibrinogênio , Ativação Plaquetária , Agregação PlaquetáriaRESUMO
Estudos publicados nas duas últimas décadas sugerem um aumento do risco de doença cardiovascular (DCV) em pacientes com periodontite, mas os mecanismos fisiopatológicos dessa associação ainda não estão completamente esclarecidos. Uma vez que foi demonstrado aumento da ativação plaquetária e do estresse oxidativo na periodontite, o objetivo do presente estudo foi investigar a via L-arginina-óxido nítrico (NO)- guanosina monofosfato cíclica (GMPc) e parâmetros de estresse oxidativo em plaquetas de pacientes com periodontite, bem como avaliar o efeito do tratamento periodontal não-cirúrgico nessas variáveis. Um total de 10 pacientes sem periodontite (periodontalmente saudáveis ou com gengivite) e 10 pacientes com periodontite participaram do estudo. A avaliação clínica, laboratorial e experimental foi realizada no início do estudo e 90 dias após realização da terapia periodontal básica (grupo periodontite). A avaliação clínica periodontal incluiu registros de: profundidade de bolsa à sondagem (PBS), nível de inserção (NIC), percentual de placa e percentual de sangramento à sondagem. Os seguintes experimentos foram realizados: influxo de L-arginina; atividade e expressão das enzimas óxido nítrico sintase e da arginase; expressão das enzimas guanilato ciclase solúvel e fosfodiesterase 5; determinação dos níveis intraplaquetários de GMPc; agregação plaquetária; avaliação do estresse oxidativo (atividade oxidante total, atividade das enzimas antioxidantes catalase e da superóxido dismutase - SOD); medição dos níveis de proteína C reativa (CRP) e de fibrinogênio. Os resultados obtidos no início do estudo demonstraram ativação do influxo de L-arginina em plaquetas via sistema y+L nos pacientes com periodontite, bem como concentrações intraplaquetárias de GMPc diminuídas e aumento sistêmico da CRP. Após o tratamento periodontal, observou-se redução do percentual de sítios com PBS ≥ 6 mm, NIC 4-5 mm e NIC ≥ 6 mm, aumento nos níveis de GMPc, para níveis ...
Studies published over the last two decades have suggested an increase of cardiovascular disease (CVD) risk on periodontitis patients, but the physiopathological mechanisms involved in this association are not yet clear. Since it has been demonstrated an enhancement on both platelet activation and oxidative stress on periodontitis patients, the aim of this study was to investigate the L-arginine-nitric oxide (NO)-cyclic guanosine monophosphate (cGMP) pathway on platelets from periodontitis patients, and the effect of non-surgical periodontal treatment in these variables. A total of 10 patients without periodontitis (periodontal healthy controls or gingivitis patients) and 10 periodontitis patients were included in this study. The clinical, laboratorial, and experimental evaluations were performed at the beginning of the study and 90 days after the basic periodontal therapy (periodontitis group). The clinical periodontal evaluation included the measurements of probing pocket depth (PPD), clinical attachment level (CAL), plaque percentage, and percentage of bleeding on probing. The following experiments were performed: L-arginine influx; nitric oxide synthase and arginase enzymes activity and expression; expression of guanylate cyclase and phosphodiesterase-5 enzymes; measurement of intraplatelet cGMP levels; platelet aggregation; oxidative stress evaluation (total oxidant activity and activity of both antioxidant enzymes catalase and superoxide dismutase SOD); measurement of C reactive protein (CRP) and fibrinogen. The initial results demonstrated an activation of L-arginine influx in platelets from periodontitis patients via y+L system, reduced intraplatelet cGMP levels and increased CRP. After periodontal treatment, it was observed reduction on percentage of sites with PPD ≥ 6 mm, CAL 4-5 mm and CAL ≥ 6 mm, enhancement on cGMP levels, to levels comparables to patients without periodontitis, accompanied by a higher activity of both antioxidant ...
Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Arginina/metabolismo , GMP Cíclico/metabolismo , Estresse Oxidativo , Óxido Nítrico/metabolismo , Periodontite/terapia , Plaquetas/enzimologia , Plaquetas/metabolismo , Proteína C-Reativa , Doenças Cardiovasculares/etiologia , Fibrinogênio , Ativação Plaquetária , Agregação PlaquetáriaRESUMO
Nitric oxide (NO) is a short-lived intercellular messenger with multiple biological implications, such as regulation of blood pressure, inhibition of platelet adhesion and aggregation, bacterial-challenge and cytokine stimulation, and regulation of mineralized tissue function. NO synthase (NOS) catalyses the conversion of cationic amino acid L-arginine to L-citrulline and NO. Recently there is an increasing interest in the role of NO in the physiopathology of periodontal disease (PD). PD is a chronic inflammatory disease of the attachment structures of the teeth, which is found in 40-50% of most adult populations worldwide and may result in tooth loss. The potential sources of NO in periodontum are inflammatory cells, keratinocytes, fibroblasts, osteoclastics and blood vessels. Etiological periodontitis factors, such as inflammatory cytokines and periodontopathogens are evolved in enhanced NO levels, which may be part of a nonspecific natural defense mechanism or may lead to periodontal damage. This review gives detail of recent research data focusing on NO bioavailability and its involvement in periodontitis pathogenesis and the modulation of NO for better control of this disease.