ABSTRACT
This study aimed to isolate, characterize chemical-structurally and evaluate the effects of polysaccharides from Caesalpinia (Libidibia) ferrea stem barks in the haemostatic system. The deproteinated-polysaccharide extract (PE-Cf) after being fractionated by ion exchange chromatography-DEAE-cellulose resulted in three fractions (FI, FII, FIII) containing total carbohydrates (14.3-38%), including uronic acid (5-16%), and polyphenols (0.94-1.7 mg/g GAE). The polysaccharide fractions presented polydisperse profile in polyacrylamide gel electrophoresis (detected by Stains-All) and molecular masses (9.5 × 104 Da-1.5 × 105 Da) identified by gel permeation chromatography. FT-IR showed absorption bands (1630 cm-1, 1396-1331 cm-1), indicative of uronic acid, and a band at 1071 cm-1, typical of COO- groups of galacturonic acid. The NMR spectra of C. ferrea polysaccharides revealed a central core composed mainly by 5-linked α-Araf and minority components as α-Rhap and α-GalAp. UV spectra of fractions revealed discrete shoulders at 269-275 nm, characteristic of polyphenolic compounds. In vitro, polysaccharides inhibited the intrinsic and/or common coagulation pathway (aPTT test) (2.0-3.7 fold) and the platelet aggregation induced by 3 µM adenosine diphosphate (25-48%) and 5 µg/mL collagen (24%), but not that induced by arachidonic acid. In vivo, the polysaccharides inhibited (36-69%) venous thrombosis induced by hypercoagulability and stasis, showing discrete hemorrhagic effect. In conclusion, the polysaccharides of C. ferrea barks, containing arabinose, galactose, rhamnose and uronic acid, possess anticoagulant, antiplatelet and antithrombotic properties of low hemorrhagic risk, suggesting potential applicability in thromboembolic disorders.
Subject(s)
Caesalpinia/metabolism , Polysaccharides/chemistry , Polysaccharides/isolation & purification , Animals , Anticoagulants/chemistry , Blood Coagulation/drug effects , Fibrinolytic Agents/chemistry , Humans , Partial Thromboplastin Time , Plant Bark/chemistry , Plant Extracts/pharmacology , Platelet Aggregation/drug effects , Polyphenols/pharmacology , Rats , Rats, Wistar , Venous ThrombosisABSTRACT
Mimosa tenuiflora (Mimosaceae) or "jurema-preta" is well distributed in the northeast Brazil, being popularly used to treat skin lesions, burns and inflammation. The healing effect of the alcoholic extract prepared with its barks corroborates the popular use. This study aimed to evaluate the inflammatory response of polysaccharides extracted from M. tenuiflora barks (EP-Mt) by methanol/NaOH and ethanol precipitation. Inflammatory activity was assessed in rat models of acute inflammation (paw edema and peritonitis), by the following parameters: edema, vascular permeability, leukocyte migration, myeloperoxidase activity and pharmacological modulation of nitric oxide and prostaglandins. EP-Mt presented 3.8% yield, 41% carbohydrate and 0.34% protein. EP-Mt (0.01, 0.1, 1.0 mg kg-1) injected by subcutaneous route elicited paw edema that lasted from 30-420 min, with maximal effect at 1 mg kg-1 (40x vs. saline), and was inhibited by L-NAME (52%) and dexamethasone (26%). EP-Mt (1 mg kg-1, via intraperitoneal) stimulated leukocytes migration (2.2x), mainly neutrophils (6.5x) and MPO activity (96%). The leukocyte migration elicited by EP-Mt was inhibited by dexamethasone (39%) and L-NAME (38%). EP-Mt containing high carbohydrate content induces acute inflammation via nitric oxide, which open perspectives of application in pathological conditions of immunosuppression.
Mimosa tenuiflora (Mimosaceae) ou "jurema-preta", amplamente distribuída no nordeste brasileiro, é utilizada popularmente no tratamento de lesões de pele, queimaduras e inflamação. O efeito cicatrizante do extrato alcoólico de suas cascas corrobora o uso popular. Avaliou-se o efeito inflamatório dos polissacarídeos da casca de M. tenuiflora (EP-Mt), obtidos por extração com metanol/NaOH e precipitação com etanol. O efeito inflamatório foi avaliado em modelos agudos em ratos (edema de pata, peritonite) por meio dos seguintes parâmetros: edema, permeabilidade vascular, migração leucocitária, atividade da mieloperoxidase e modulação farmacológica de prostaglandinas e óxido nítrico. EP-Mt apresentou 3,8% de rendimento, 41% de carboidratos totais e 0,34% de proteína. EP-Mt (0,01, 0,1, 1,0 mg kg -1), administrado por via subcutânea, induziu edema de pata com duração de 30 a 420 min e efeito máximo na dose de 1 mg kg-1 (40x vs. salina), o qual foi inibido por L-NAME (52%) e dexametasona (26%). EP-Mt (1 mg kg-1, via intraperitoneal) estimulou a migração de leucócitos (2,2x vs. salina), principalmente de neutrófilos (6,5x), com aumento da atividade da mieloperoxidase (96%). A migração de leucócitos foi inibida por dexametasona (39%) e L-NAME (38%). EP-Mt contendo elevado teor de carboidratos induz inflamação aguda via óxido nítrico com perspectivas de aplicação em condições patológicas de imunossupressão.
Subject(s)
Rats , Carbohydrates , Inflammation , Mimosa , Plants, MedicinalABSTRACT
The South American giant ant, Dinoponera quadriceps (Hymenoptera, Formicidae, Ponerinae), produces proteinaceous venom that has antinociceptive, neuroprotective and antimicrobial effects, thereby supporting the popular use of these ants to treat asthma, rheumatism, earache and back pain. Anticoagulant activity is another biological property that has been shown for the venom of other hymenopteran species, like wasps. The aim of this study was to assess the anti-inflammatory, anticoagulant and antiplatelet properties of D. quadriceps venom (DqV). DqV anti-inflammatory activity was assessed by intravenous administration in Swiss mice in the models of paw edema and peritonitis. In vitro, DqV was assessed in coagulation (activated partial thromboplastin time) and platelet aggregation tests. DqV inhibited (27-33%) the edema elicited by carrageenan and the leucocyte migration (43%) elicited by zymosan. DqV decreased by 57% and 42%, respectively, the content of malondialdehyde and nitrite in the peritoneal fluid. DqV prolonged (1.8x) the clotting time and decreased (27%) the platelet aggregation induced by adenosine diphosphate. The crude venom of D. quadriceps presents an anti-inflammatory effect in mice and in vitro anticoagulant and antiplatelet effects.
