ABSTRACT
Objective·To isolate lipopolysaccharide (LPS) and capsular polysaccharide (CPS) from Porphyromonas gingivalis (Pg) and analyze their ability to induce THP-1 cells to secrete interleukin 8 (IL-8) in vitro. Methods·LPS and CPS were isolated from the polysaccharides by gel filtration chromatography, which had already been extracted from Pg ATCC33277 and SJD4. The isolated LPS and CPS from different strains were used to stimulate THP-1 cells in different doses and for different periods. The level of IL-8 in the cell culture suspension was measured by ELISA. Results·LPS and CPS were successfully separated from the polysaccharides in Pg ATCC33277 and SJD4. The relative molecular masses of LPS and CPS were 20000-50000 and 150000-500000, respectively. LPS and CPS could induce THP-1 cells to produce IL-8 in vitro in a time- and dose-dependent manner. The effect of LPS from ATCC33277 on secretion of IL-8 was greater than that of CPS with the dose of 0.1 μg/mL (P<0.05). However, the difference could only be found at 24 h in SJD4, and the effect of CPS was greater (P<0.05). Conclusion·As a component of polysaccharides extracted from Pg, CPS can induce THP-1 cells to secrete IL-8 in vitro in a time- and dose-dependent manner.
ABSTRACT
RACIONAL: A videocirurgia pode apresentar complicações inerentes ao método e dentre elas destacam-se as relacionadas ao aumento da pressão na cavidade intra-abdominal. OBJETIVO: Analisar os efeitos do pneumoperitônio em modelo experimental de endotoxemia causada por lipopolissacarídeo. MÉTODO: Foram utilizados 32 ratos Wistar dos quais foi coletado sangue 24 horas prévias para obtenção dos valores de referência nas provas de função renal, hepática e do estado endotóxico (contagens total e diferencial de leucócitos, contagem de plaquetas e dosagens de lipopolissacarídeo, TNFα, IL6). A seguir os ratos foram separados em quatro grupos com oito: grupos controle inoculados com lipopolissacarídeo (10 mg/kg via intraperitoneal) e mantidos por quatro e 11 horas (C-LPS4 e C-LPS11). Grupo C-PP, submetido ao pneumoperitônio por CO2 por uma hora e mantido sob observação por seis horas. Grupo experimento (E) inoculados com 10 mg/kg de lipopolissacarídeo por via intraperitoneal, após quatro horas submetidos ao pneumoperitônio por CO2 por uma hora e mantidos sob observação por seis horas. Ao término destes períodos foram coletadas amostras de sangue para as mesmas avaliações iniciais. Utilizou-se o método de T-Student para as comparações dos resultados, com nível de significância de 0,05. RESULTADOS: Em comparação aos valores iniciais e aos grupos controle, o grupo E apresentou maiores valores nas dosagens de creatinina e uréia, bilirrubina direta, ALT e TAP, aumento nas dosagens de IL6, TNFα e diminuição de plaquetas, aumento de leucócitos e bastonetes e detecção de endotoxina circulante. CONCLUSÃO: O pneumoperitônio por dióxido de carbono induz aumentos na liberação de TNFα, IL6, piora das funções hepática e renal em modelo experimental de endotoxemia induzida por lipopolissacarídeo.
BACKGROUND: Videosurgery can bring complications with it´s manipulation and among them the hyperpression into the abdominal cavity is important factor. AIM: To analyze the pneumoperitoneum effects in an endotoxemia experimental model caused by lipopolyssacharide. METHOD: Thirty-two Wistar rats were utilized, from which blood was collected 24 hours prior to the experiment for the initial evaluations of the renal function, the hepatic function and the endotoxic state (total counts and differential of leukocytes, platelet count and dosages of lipopolyssacharide, TNFα, IL6). The rats were then separated into four groups of eight: control groups inoculated with lipopolyssacharide (10 mg/kg via intraperitoneum) and kept for four and 11 hours (C-LPS4 and C-LPS11). Group C-PP, control of pneumoperitoneum by carbon dioxide at 10 mmHg for one hour and kept under observation for six hours. Experiment group (E) in which the rats were inoculated with 10 mg/kg of lipopolyssacharide via intraperitoneum and after four hours were submitted to pneumoperitoneum by carbon dioxide at 10 mmHg for one hour and kept under observation for six hours. At the end of these periods, blood samples were collected for the same initial evaluations. The T-student method was utilized for statistical comparisons of results, with a significance level of 0.05. RESULTS: Compared to the initial values and the control groups, Group E had higher values in the renal function tests (creatinine=0.85 ± 0.24 and urea=106.38 ± 37.61 mg/dL). In the hepatic function tests, there were higher values in the dosages of direct bilirubin (0.34 ± 0.09mg/dL), ALT (390,38 ± 351.88) and TAP (17.01 ± 2.18). The endotoxic state was confirmed by the dosages of IL6 (36.0 ± 11.23 pg/mL), TNFα (518.36 ± 203.39 pg/mL) and by the hematological alterations: reduction of platelets, increased leukocytes and band neutrophils and the detection of circulating endotoxin (0.21 ± 0.08 UE/mL). CONCLUSION...