RESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Remirea maritima Aubl., popularly known as "capim-da-praia", is popularly employed in the treatment of diarrhea, kidney disease, fever, and for analgesic and anti-inflammatory purposes through the preparation of teas. Few studies have focused on the chemical composition and its biological properties. AIM OF THE STUDY: This work evaluated the antinocipetive, anti-inflammatory and antioxidant activities of the aqueous extract from Remirea maritima Aubl. as well as the isolation and identification of the chemical compounds. MATERIALS AND METHODS: Compounds were isolated from aqueous extract of Remirea maritima through preparative HPLC and the structures were identified by means of NMR and MS analysis. The tests for antinociceptive, anti-inflammatory, and antioxidant activities, along with motor coordination test (Rota rod), were performed over the aqueous extract. RESULTS: The phytochemical investigation of aqueous extract of Remirea maritima resulted in the isolation of three flavone glycosides. The structures of these compounds were determined by means of MS and 1D and 2D NMR data as vitexin-2â³-O-ß-D-glucopyranoside, isovitexin-2â³-O-ß-D-glucopyranoside, and luteolin-7-O-glucuronide. Acute pretreatment with aqueous extract (100, 200 or 400mg/kg, i.p.) caused a significant decrease (p<0.001) in the number of abdominal writhes. In the formalin test, higher doses significantly inhibited the late (inflammatory pain) phase of formalin-induced licking (p<0.05 or 0.001). In the hot plate test, there was no significant difference in nociceptive behavior, discarding the possible central effect of the aqueous extract. In the rota rod test, it was verified that the aqueous extract in all concentration evaluated does not alter the motor coordination of mice, such antinociceptive results were unlikely to be caused by motor abnormality. In the peritonitis test, induced by carrageenan, the treatment with aqueous extract produced a significant reduction in leukocyte migration in all concentration evaluated. Additionally, a significant reduction of lipoperoxidation (TBARS test) and in nitric oxide formation (.NO Scavenging assay) was observed in antioxidant activity assay. CONCLUSION: The biological and phytochemical investigations of the aqueous extract of Remirea maritima resulted in the identification of three flavone glycosides that have been described here for the first time in Remirea and effective analgesic activity in various pain models, probably mediated via the inhibition of peripheral mediators which could be related to its strong antioxidant effect observed in vitro.
Assuntos
Analgésicos/uso terapêutico , Anti-Inflamatórios/uso terapêutico , Antioxidantes/uso terapêutico , Fitoterapia/métodos , Extratos Vegetais/uso terapêutico , Analgésicos/química , Analgésicos/farmacologia , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Carragenina , Cyperaceae/química , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Flavonas/química , Flavonas/farmacologia , Glicosídeos/química , Glicosídeos/farmacologia , Masculino , Camundongos , Estrutura Molecular , Medição da Dor/efeitos dos fármacos , Peritonite/induzido quimicamente , Peritonite/tratamento farmacológico , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Teste de Desempenho do Rota-Rod/métodosRESUMO
The objectives of the present study were the subtyping of Campylobacter jejuni subsp. jejuni strains obtained from humans and different animal species using PCR-RFLP, and the detection, by means of the same technique, of strains related to serotype PEN O19:LIO 7, the main C. jejuni serotype linked to Guillain-Barré Syndrome (GBS). Seventy C. jejuni strains isolated from human feces (n=33), primates (n=15), dogs (n=5), swine (n=2), bovines (n=1), abortion material from goats (n=2) and poultry carcasses (n=12), all collected in the state of São Paulo, were subtyped by means of PCR-RFLP of fla A gene, using restriction endonucleases Hae III, Afa I and Mbo I. Seven subtypes were observed when using the enzyme Hae III; eight when using Mbo I; and seven when using Afa I. The combination of the three endonucleases led to 16 fla-RFLP subtypes, from which ten subtypes shared strains of human and animal origin. From these, seven subtypes were observed in human and broiler strains. In eight subtypes, the other animal species shared patterns with human strains. It was inferred that, besides broilers, swine, goats, dogs and primates may be sources of infection for human in São Paulo. PCR-RFLP is a highly discriminatory technique that may be applied to molecular epidemiology studies of samples from different origins. Besides, the study also enabled the detection of two human strains and two primate strains related to serotype PEN O19: LIO 7.
Assuntos
Humanos , Animais , Infecções por Campylobacter , Campylobacter jejuni/isolamento & purificação , Técnicas e Procedimentos Diagnósticos , Técnicas In Vitro , Polimorfismo de Fragmento de Restrição , Reação em Cadeia da Polimerase/métodos , Síndrome de Guillain-Barré/diagnóstico , Estudos Epidemiológicos , Métodos , Estudos de Amostragem , MétodosRESUMO
The objectives of the present study were the subtyping of Campylobacter jejuni subsp. jejuni strains obtained from humans and different animal species using PCR-RFLP, and the detection, by means of the same technique, of strains related to serotype PEN O19:LIO 7, the main C. jejuni serotype linked to Guillain-Barré Syndrome (GBS). Seventy C. jejuni strains isolated from human feces (n=33), primates (n=15), dogs (n=5), swine (n=2), bovines (n=1), abortion material from goats (n=2) and poultry carcasses (n=12), all collected in the state of São Paulo, were subtyped by means of PCR-RFLP of fla A gene, using restriction endonucleases Hae III, Afa I and Mbo I. Seven subtypes were observed when using the enzyme Hae III; eight when using Mbo I; and seven when using Afa I. The combination of the three endonucleases led to 16 fla-RFLP subtypes, from which ten subtypes shared strains of human and animal origin. From these, seven subtypes were observed in human and broiler strains. In eight subtypes, the other animal species shared patterns with human strains. It was inferred that, besides broilers, swine, goats, dogs and primates may be sources of infection for human in São Paulo. PCR-RFLP is a highly discriminatory technique that may be applied to molecular epidemiology studies of samples from different origins. Besides, the study also enabled the detection of two human strains and two primate strains related to serotype PEN O19: LIO 7.
RESUMO
Ampelozizyphus amazonicus Ducke is a tree commonly found in the Amazon region and an extract of its stem bark is popularly used as an antimalarial and anti-inflammatory agent and as an antidote to snake venom. Ursolic acid; five lupane type triterpenes: betulin, betulinic acid, lupenone, 3ß-hydroxylup-20(29)-ene-27,28-dioic acid, and 2a,3ß-dihydroxylup-20(29)-ene-27,28-dioic acid, and three phytosteroids: stigmasterol, sitosterol and campesterol, have been isolated from stem extracts of A. amazonicus Ducke. Their structures were characterized by spectral data including COSY and HMQC. In an in vitro biological screening of the isolated compounds, 3ß-hydroxylup-20(29)-ene-27,28-dioic acid was cytotoxic against the SKBR-3 human adenocarcinoma cell line (1 to 10 mg/mL), while 2a,3ß-dihydroxylup-20(29)-ene-27,28-dioic acid exhibited cytotoxicity against both SKBR-3 human adenocarcinoma and C-8161 human melanoma tumor cell lines (>0.1 mg/mL). In the present study, different extracts and some fractions of this plant were also investigated for trypanocidal activity due to the presence of pentacyclic triterpenes. The triterpene classes are potent against Trypanosoma cruzi. The bioassays were carried out using blood collected from Swiss albino mice by cardiac puncture during the parasitemic peak (7th day) after infection with the Y strain of T. cruzi. The results obtained showed that A. amazonicus is a potential source of bioactive compounds since its extracts and fractions isolated from it exhibited in vitro parasite lysis against trypomastigote forms of T. cruzi at concentrations >100 æg/mL. Fractions containing mainly betulin, lupenone, 3ß-hydroxylup-20(29)-ene-27,28-dioic acid, and 2a,3ß-dihydroxylup-20(29)-ene-27,28-dioic acid showed more activity than crude extracts.
Assuntos
Animais , Camundongos , Rhamnaceae/química , Tripanossomicidas/farmacologia , Trypanosoma cruzi/efeitos dos fármacos , Linhagem Celular Tumoral/efeitos dos fármacos , Extratos Vegetais/farmacologia , Tripanossomicidas/química , Tripanossomicidas/isolamento & purificaçãoRESUMO
Ampelozizyphus amazonicus Ducke is a tree commonly found in the Amazon region and an extract of its stem bark is popularly used as an antimalarial and anti-inflammatory agent and as an antidote to snake venom. Ursolic acid; five lupane type triterpenes: betulin, betulinic acid, lupenone, 3beta-hydroxylup-20(29)-ene-27,28-dioic acid, and 2alpha,3beta-dihydroxylup-20(29)-ene-27,28-dioic acid, and three phytosteroids: stigmasterol, sitosterol and campesterol, have been isolated from stem extracts of A. amazonicus Ducke. Their structures were characterized by spectral data including COSY and HMQC. In an in vitro biological screening of the isolated compounds, 3beta-hydroxylup-20(29)-ene-27,28-dioic acid was cytotoxic against the SKBR-3 human adenocarcinoma cell line (1 to 10 mg/mL), while 2alpha,3beta-dihydroxylup-20(29)-ene-27,28-dioic acid exhibited cytotoxicity against both SKBR-3 human adenocarcinoma and C-8161 human melanoma tumor cell lines (>0.1 mg/mL). In the present study, different extracts and some fractions of this plant were also investigated for trypanocidal activity due to the presence of pentacyclic triterpenes. The triterpene classes are potent against Trypanosoma cruzi. The bioassays were carried out using blood collected from Swiss albino mice by cardiac puncture during the parasitemic peak (7th day) after infection with the Y strain of T. cruzi. The results obtained showed that A. amazonicus is a potential source of bioactive compounds since its extracts and fractions isolated from it exhibited in vitro parasite lysis against trypomastigote forms of T. cruzi at concentrations >100 microg/mL. Fractions containing mainly betulin, lupenone, 3beta-hydroxylup-20(29)-ene-27,28-dioic acid, and 2alpha,3beta-dihydroxylup-20(29)-ene-27,28-dioic acid showed more activity than crude extracts.
Assuntos
Rhamnaceae/química , Tripanossomicidas/farmacologia , Trypanosoma cruzi/efeitos dos fármacos , Animais , Linhagem Celular Tumoral/efeitos dos fármacos , Camundongos , Extratos Vegetais/farmacologia , Tripanossomicidas/química , Tripanossomicidas/isolamento & purificaçãoRESUMO
We present here a retrospective study of nine patients diagnosed with Whipple's disease (WD) in our hospital. This report em-phasises clinical presentation, diagnostic methods, treatment and response to treatment. In our study, the disease was more frequent in males, and the most frequent presenting symptoms were arthralgia, diarrhoea and weight loss. Since the intestine is almost always affected, oral endoscopy is a useful technique for the diagnosis of WD because it shows the typical miliary pattern and aids in obtaining biopsies to show the presence of PAS-positive macrophages (a suggestive though not diagnostic finding), to show bacilli using electron microscopy, or to detect genetic material using PCR. Our patients responded well to treatment. The most frequently used antibiotic was oral trimethoprim-sulfamethoxazole for at least one year. Treatment with penicillin G and IM streptomycin for 14 days was reserved for severe cases or cases that responded poorly to treatment.
Assuntos
Doença de Whipple , Adulto , Idoso , Antibacterianos/administração & dosagem , Antibacterianos/uso terapêutico , Anti-Infecciosos/administração & dosagem , Anti-Infecciosos/uso terapêutico , Biópsia , Endoscopia , Feminino , Humanos , Mucosa Intestinal/patologia , Intestino Delgado/patologia , Masculino , Pessoa de Meia-Idade , Penicilina G/administração & dosagem , Penicilina G/uso terapêutico , Penicilinas/administração & dosagem , Penicilinas/uso terapêutico , Radiografia Abdominal , Estudos Retrospectivos , Estreptomicina/administração & dosagem , Estreptomicina/uso terapêutico , Fatores de Tempo , Tomografia Computadorizada por Raios X , Combinação Trimetoprima e Sulfametoxazol/administração & dosagem , Combinação Trimetoprima e Sulfametoxazol/uso terapêutico , Doença de Whipple/diagnóstico , Doença de Whipple/tratamento farmacológico , Doença de Whipple/patologiaRESUMO
Crude extracts of Leishmania amazonensis, but not of L. guyanensis, are lytic to erythrocytes and nucleated cells, including macrophages. L. amazonensis-mediated lysis is caused by a membrane-associated pore-forming protein, named a-leishporin. Here we show that L. amazonensis, but not L. guyanensis, promastigote extracts increase their hemolytic activity when kept at 4 degrees C for a few days or at 37 degrees C for a few hours. We show that the activation in the extracts is mediated by a cytosolic serine-protease. Although L. guyanensis extracts are hemolytically inactive and unable to generate hemolytic activity, their membrane fraction becomes hemolytic in the presence of the cytosolic fraction of L. amazonensis, also by the action of a serine-protease. This suggests that L. guyanensis contains a potential lytic molecule, named here g-leishporin. The cytosolic fraction of L. guyanensis is unable to activate either a- or g-leishporin, indicating that this species does not possess the protease(s) that activate(s) the cytolysin. Trypsin, chymotrypsin, collagenase, Pronase and proteinase K, are also effective in activating a-leishporin but not g-leishporin. This suggests that the inactive forms of a-leishporin and g-leishporin are distinct in structure and/or are activated by different mechanisms. We are considering two hypotheses for the activation of leishporins: (1) proteolysis of an inactive precursor and (2) dissociation and/or proteolytic degradation of an inhibitory oligopeptide. The present data and preliminary results argue for the second hypothesis. We speculate that leishporin could be activated in the protease-rich, low pH, and dissociating environment of parasitophorous vacuole contributing for the release of the parasites from the macrophage.
Assuntos
Citotoxinas/metabolismo , Eritrócitos/fisiologia , Hemólise , Leishmania/metabolismo , Proteínas de Protozoários/metabolismo , Serina Endopeptidases/metabolismo , Animais , Citosol/metabolismo , Ativação Enzimática , Humanos , Concentração de Íons de Hidrogênio , Leishmania guyanensis/metabolismo , TemperaturaRESUMO
During their growth in vitro, promastigotes of Leishmania amazonensis undergo differentiation from complement-susceptible to complement-resistant forms. Here, we demonstrate that both forms bind comparable amounts of C3 on their surfaces, with the predominant molecule species being the haemolytically active C3b. Likewise, equivalent amounts of C9 are deposited on both forms of promastigotes. However, while C9-bearing complexes are exposed on the cell surface of resistant promastigotes, they are cryptic in the susceptible stage of the parasites. The membrane fraction of complement-resistant promastigote lysates has the ability to inhibit complement-mediated haemolysis, blocking C9, but not C3 deposition to complement-activating complexes. Moreover, the membrane fraction of complement-resistant promastigote lysates can inhibit the late steps of guinea-pig erythrocyte lysis much more efficiently than complement-susceptible ones. Our results indicate that L. amazonensis promastigotes evade complement killing by inhibiting the cytolytic pathway of the complement cascade.