RESUMO
Rosmarinus officinalis L. (Lamiaceae), popularly known as rosemary, is used for food flavoring and in folk medicine as an antispasmodic, analgesic, antirheumatic, diuretic, and antiepileptic agent. Few studies have shown the anti-inflammatory effects of rosemary essential oil (REO). This study evaluated the effects of REO on leukocyte migration through in vivo leukocyte migration and in vitro chemotaxis assay. REO was analyzed by using gas chromatography-mass spectometry, and the main components identified were camphor (27.59%), 1,8-cineole (15.74%), α-pinene (16.58%), and ß-myrcene (10.02%). In rats, administration of REO reduced the number of leukocytes that rolled, adhered, and migrated to the scrotal chamber after carrageenan injection. All doses of REO tested significantly inhibited leukocyte chemotaxis induced by casein. The effects of REO on leukocyte migration highlight an important mechanism of the anti-inflammatory action of rosemary.
Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Inibição de Migração Celular , Quimiotaxia de Leucócito/efeitos dos fármacos , Leucócitos/efeitos dos fármacos , Óleos Voláteis/farmacologia , Animais , Anti-Inflamatórios não Esteroides/administração & dosagem , Anti-Inflamatórios não Esteroides/química , Carragenina/toxicidade , Ensaios de Migração de Leucócitos , Relação Dose-Resposta a Droga , Endotélio/citologia , Endotélio/efeitos dos fármacos , Endotélio/imunologia , Cromatografia Gasosa-Espectrometria de Massas , Teste de Inibição de Aderência Leucocítica , Leucócitos/citologia , Leucócitos/imunologia , Espectroscopia de Ressonância Magnética , Masculino , Medicina Tradicional , Monoterpenos/análise , Monoterpenos/química , Monoterpenos/farmacologia , Óleos Voláteis/administração & dosagem , Óleos Voláteis/química , Folhas de Planta/química , Ratos , Ratos Wistar , Rosmarinus/química , Escroto/citologia , Escroto/efeitos dos fármacos , Escroto/imunologiaRESUMO
OBJECTIVES: Increased homocysteine contributes to the pathophysiology of several chronic inflammatory diseases. Whether homocysteine could participate in mucosal inflammation in inflammatory bowel disease (IBD) has not been explored yet. Our aims were to study the levels of plasma and mucosal homocysteine in IBD patients and to assess whether homocysteine can trigger an inflammatory reaction on human intestinal microvascular endothelial cells (HIMECs). METHODS: Homocysteine was measured in the plasma, mucosal biopsy, and lamina propria mononuclear cell (LPMC) supernatants from normal and IBD subjects. HIMEC were cultured in presence of homocysteine, TNF-alpha, or folic acid, alone or in combination. Expression of vascular cell adhesion molecule 1 (VCAM-1) and intercellular cell adhesion molecule 1 was measured by flow cytometry and monocyte chemoattractant protein-1 (MCP-1) production by ELISA. Phosphorylation of p38 and p42/44 was assessed by immunoblot in HIMEC extracts. T-cell- and monocyte-HIMEC adhesion assays were used to evaluate the impact of homocysteine on leukocyte adhesion to intestinal endothelial cells. RESULTS: Patients with IBD displayed significantly higher homocysteine plasma and mucosal levels than control subjects. IBD-derived LPMC released higher homocysteine than control-derived LPMC. Treatment of HIMEC with homocysteine, and synergistically with the combination of TNF-alpha and homocysteine, triggered HIMEC inflammation, resulting in VCAM-1 up-regulation, MCP-1 production, and p38 phosphorylation. These events lead to an increased capacity of HIMEC to adhere T- and monocyte cells and were blocked by folic acid treatment. CONCLUSIONS: Homocysteine is increased in both the mucosa and plasma of patients with Crohn's disease and ulcerative colitis and contributes to the inflammatory state of the mucosal IBD endothelium. Therefore, homocysteine could play a proinflammatory role in IBD, which can be efficiently targeted by folic acid supplementation.
Assuntos
Colite Ulcerativa/fisiopatologia , Doença de Crohn/fisiopatologia , Homocisteína/fisiologia , Mucosa Intestinal/irrigação sanguínea , Adolescente , Adulto , Idoso , Quimiocina CCL2/sangue , Endotélio Vascular/fisiopatologia , Ensaio de Imunoadsorção Enzimática , Feminino , Citometria de Fluxo , Ácido Fólico/sangue , Humanos , Molécula 1 de Adesão Intercelular/sangue , Teste de Inibição de Aderência Leucocítica , Masculino , Microcirculação/fisiopatologia , Pessoa de Meia-Idade , Valores de Referência , Fator de Necrose Tumoral alfa/metabolismo , Molécula 1 de Adesão de Célula Vascular/sangueRESUMO
In order to study the influence of endothelial cell fatty acid composition on various membrane related parameters, several in vitro methods were developed for manipulating the fatty acid content of human endothelial cell membranes. Changes in membrane fatty acid profile were induced by using fatty acid modified lipoproteins or free fatty acids. The largest changes in endothelial fatty acid composition were obtained by culturing the cells in media supplemented with specific free fatty acids. An increase in arachidonic acid content of endothelial phospholipids was induced by supplementation with saturated fatty acids or with arachidonic acid itself. A decrease in arachidonic acid content was obtained by supplementation with other unsaturated fatty acids. Under the experimental conditions used endothelial cells showed a low desaturase activity and a high elongase activity. Considerable alterations in membrane fatty acid composition did not greatly influence certain membrane related parameters such as polymorphonuclear leukocyte adherence and endothelial cell procoagulant activity. In general, for fatty acid modified endothelial cells an association between endogenous arachidonic acid content and total production of eicosanoids was found. This study demonstrates that considerable changes in membrane fatty acid profile affect endothelial cell arachidonic acid metabolism, but it also illustrates homeostasis at the level of endothelial cell functional activity.
Assuntos
Membrana Celular/química , Endotélio Vascular/química , Ácidos Graxos/análise , Fatores de Coagulação Sanguínea/análise , Adesão Celular , Células Cultivadas , Meios de Cultura , Ácidos Eicosanoicos/análise , Endotélio Vascular/fisiologia , Ácidos Graxos/farmacologia , Humanos , Teste de Inibição de Aderência Leucocítica , Neutrófilos/fisiologia , Ácido Oleico , Ácidos Oleicos/análise , Fosfolipídeos/análise , Tromboplastina/análiseRESUMO
A cohort of nine extrinsic asthma patients were treated by means of acupuncture. Patients were followed up for changes in medical treatment, spirometry, skin reactivity to immediate type reactions, total serum IgE levels and reactivity of their leukocytes to leukotriene C4 challenge (LTC4 induced leukocyte adherence inhibition (LAI) assay). Our results show that after acupuncture, treated patients were able to reduce bronchodilator and taper completely corticosteroid therapy. No change in skin reactivity or in IgE levels were noted. However, acupuncture treatment was able to negate, in 66.6%, the positive LTC4 induced responses.
Assuntos
Terapia por Acupuntura/métodos , Asma/terapia , Leucócitos/imunologia , SRS-A , Adulto , Resistência das Vias Respiratórias , Asma/imunologia , Asma/fisiopatologia , Broncodilatadores/administração & dosagem , Feminino , Seguimentos , Humanos , Imunoglobulina E/análise , Teste de Inibição de Aderência Leucocítica , Masculino , Testes de Função Respiratória , SRS-A/imunologia , Testes CutâneosRESUMO
Lipopolysaccharides (LPS) were extracted from seven type strains of Campylobacter jejuni and one type strain of Campylobacter coli with 45 per cent aqueous phenol. The sugar components present in all LPS were glucose, galactose, L-glycero-D-manno-heptose, 2-keto-3-deoxy-octulosonic acid and glucosamine. All but one LPS contained galactosamine, and two strains contained in addition mannose. The fatty acids present were 3-hydroxy-tetradecanoic acid, n-hexadecanoic acid and trace amounts of n-tetradecanoic acid. The LPS preparations examined showed anti-complementary effect, and were able to gelatinize Limulus amoebocyte lysate. LPS of the C.jejuni strain tested (NCTC 11168) was found to be lethal for mice and to produce the local Shwartzman reaction in rabbits.
Assuntos
Campylobacter/imunologia , Lipopolissacarídeos/isolamento & purificação , Animais , Carboidratos/análise , Cromatografia Gasosa , Cromatografia em Papel , Proteínas do Sistema Complemento/imunologia , Ácidos Graxos/análise , Feminino , Imunodifusão , Teste de Inibição de Aderência Leucocítica , Lipopolissacarídeos/toxicidade , Masculino , Camundongos , Camundongos Endogâmicos , Coelhos , Sorotipagem , Especificidade da EspécieRESUMO
In vitro antitumor immunity, measured by means of a tube leukocyte adherence inhibition (LAI) assay, was able to demonstrate early colorectal cancer in cases yet undiagnosed. Testing 106 patients attending a colonoscopy clinic, we found that nine of 15 (60%) patients with colorectal cancer were LAI-positive. The nine included: five of six with Dukes' A and B; one of two with Dukes' C; and three of seven with Dukes' D lesions. Of 58 patients without colon neoplasia, one had a positive test. By contrast, barium enema results were diagnostic for 13 of 15 (87%) cases and colonoscopy for 11 of 15 (73%). There were 32 colon polyps discovered by means of barium enema and colonoscopy. Of 13 adenomas removed, four had stimulated positive LAI responses. Of another 19 polyps not removed two incited positive LAI responses. Two patients with histories of recurrent adenomas were LAI-positive, though only one had a new polyp that could be detected by means of barium enema. For a procedure to be of diagnostic value for a given disease, sensitivity and specificity must be at least 1.0. If this sum is 2.0, the test is ideal. The sum of sensitivity and specificity for the tube LAI assay, barium enema, and colonoscopy were 1.58, 1.87, and 1.73, respectively, for colorectal cancer; 1.81, 2.0, and 2.0, respectively, for early colorectal cancer; and 1.42, 1.77, and 1.55 respectively, for advanced colorectal cancer. In this limited series of patients, the barium enema seems to be superior to the tube LAI assay for diagnosing colorectal cancer, but the difference in sensitivity and specificity is not statistically valid by the paired chi square test. The results of this study shows conclusively, that the tube LAI is of diagnostic value and suggest its use for evaluating suspected cases, especially early ones, of colorectal cancer.