Influência do tempo e dose de heparina em modelo de peritonite aguda

In the last years, heparin has become target of many studies related to inflammation due its ability of biding to proteins involved on immune response. Recently, it was demonstrated, at our laboratory, using a thIoglycollate-induced peritonitis model, heparin s capacity of reduce cellular influx into the peritoneal cavity, 3 hours after the inflammatory stimulus. Once neutrophilic infiltration is highest around 8 hours after the inflammatory stimulus, at the present work, using the same peritonitis model, it was assessed heparin s ability of keeping the interference on leukocyte infiltration, 8 hours after inflammation induction. Moreover, using cellular differential count, it was evaluated how the cellular populations involved in the inflammatory process would be affected by the treatment. Eight hours after the inflammatory stimulus, only heparin dosage of 1 μg/Kg was able to reduce the cellular influx to peritoneum, 62.8% of reduction when compared to positive control (p < 0.001). Furthermore, heparin dosage of 15 μg/Kg presented a pro-inflammatory effect in whole blood verified by the increase of 60.9% (p < 0.001) and 117.8% (p < 0.001) on neutrophils and monocytes proportion, respectively, when compared to positive control. In addition, this dosage also presented a neutrophilic proportion on peritoneal fluid 27.3% higher than positive control (p < 0.05). This duality between anti- and pro-inflammatory effects at different times corroborates studies that attribute a pleiotropic immunomodulator role to heparin.

Avaliação do potencial anti-inflamatório de composto tipo heparina (cCTH) extraído do caranguejo Goniopsis cruentata em modelo experimental de peritonite

Heparin, a sulfated polysaccharide, was the first compound used as an anticoagulant and antithrombotic agent. Due to their structural characteristics, also has great potential anti-inflammatory, though such use is limited in inflammation because of their marked effects on coagulation. The occurrence of heparin-like compounds that exhibit anticoagulant activity decreased in aquatic invertebrates, such as crab Goniopsis cruentata, sparked interest for the study of such compounds as anti-inflammatory drugs. Therefore, the objective of this study was to evaluate the potential modulator of heparin-like compound extracted from Goniopsis cruentata in inflammatory events, coagulation, and to evaluate some aspects of its structure. The heparin-type compound had a high degree of N-sulphation in its structure, being able to reduce leukocyte migration into the peritoneal cavity at lower doses compared to heparin and diclofenac sodium (anti-inflammatory commercial). Furthermore, it was also able to inhibit the production of nitric oxide and tumor necrosis factor alpha by activated macrophages, inhibited the activation of the enzyme neutrophil elastase in low concentrations and showed a lower anticoagulant effect in high doses as compared to porcine mucosal heparin. Because of these observations, the compound extracted from crab Goniopsis cruentata can be used as a structural model for future anti-inflammatory agents

Efeito de extratos da alga marinha caulerpa mexicana em modelos de inflamação em murinos

The regulation of the inflammatory response is essential to maintain homeostasis. Several studies have been performed to search new drugs that can contribute to avoiding or minimizing an excessive inflammatory process. The aim of this study was to evaluate the effect of extracts of green algae Caulerpa mexican in models of inflammation. In mice, the model of peritonitis induced inflammatory zymosan pretreatment of mice with aqueous and methanol extracts of C. mexican was able to reduce cell migration to the peritoneal cavity. Treatment of mice with extracts of C. mexican also reduced the ear edema induced by xylene and exerted inhibitory action on the migration of leukocytes in inflammation-induced zymosan the air pouch, and timedependent for the extracts tested in the model of ulcerative colitis induced by DSS 3%, the extract methanol, but not the aqueous C. mexican, significantly reduced the clinical symptoms of colitis, as well as the production of proinflammatory cytokines in the culture of mouse colon, in the histological analysis there was a slight reduction of inflammation in the intestinal mucosa. We concluded that the administration of the extracts resulted in the reduction of cell migration to different sites as well as reducing the edema formation induced by chemical irritant. This study demonstrates for the first time the antiinflammatory effect of aqueous and methanolic extracts from green marine algae Caulerpa mexican

Atividade antiinflamatória de uma heterofucana da alga marrom Padina gymnospora

Fucans, sulfated polysaccharides extracted from brown algae and some echinoderms, have been extensively studied for its diverse biological activities and because of its interference with molecular mechanisms of cell to cell recognition, including leukocyte trafficking from blood vessels into sites of inflammation mediated by selectin, a family of adhesion molecules. In the present study, we examined structural features of a heterofucan extracted from brown algae Padina gymnospora and its effect on the leukocyte migration to the peritoneum. The sulfated polysaccharides were extracted from the brown seaweed by proteolysis with the proteolytic enzyme maxatase. The presence of protein and uronic acid contamination was detected in the crude polysaccharide extract. Fractionation of the crude extract with growing concentrations of acetone produced five fractions with different concentrations of fucose, xylose, uronic acid, galactose, glucose and sulfate. The fraction precipitated with 1.5 volumes of acetone was characterized by infrared and nuclear magnetic resonance, through which can be observed the presence of sulfate groups in the C4 of -L-fucose. The anti-inflammatory action of this composite was assessed by a sodium thioglycollate-induced peritonitis assay and through nitric oxide production by the peritoneal macrophages using Griess reagent. Fraction F1.5 was efficient in reducing leukocyte influx into the peritoneal cavity when 10 mg/kg and 25mg/kg were used, resulting in a decrease of 56 and 39%, respectively. A decrease of nitric oxide production occurred when high concentrations of fucana were used. The cytotoxicity of the composite was also assessed using the reduction of 3-(4,5 dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT). Fraction F1.5 had no cytotoxicity when 500 μg/mL of the fraction was used. This study suggests the use of fraction F1.5 (heterofucan) as an anti-inflammatory

Análise estrutural e avaliação do efeito condroitim sulfato extraído de tilápia (Oreochromis niloticus) em modelo de peritonite aguda

Chondroitin sulfate (CS) is a naturally glycosaminoglycan found in the extracellular matrix of connective tissues and it may be extracted and purified those tissues. CS is involved in various biological functions, which may be related to the having structural variability, despite the simplicity of the linear chain structure from this molecule. Researches in biotechnology and pharmaceutical field with wastes from aquaculture has been developed in Brazil. In recent decades, tilapia (Oreochromis niloticus), native fish from Africa, has been one of the most cultivated species in various regions of the world, including Brazil. The tilapia farming is a cost-effective activity, however, it generates large amount of wastes that are discarded by producers. It is understood that waste from tilapia can be used in research as a source of molecules with important biotechnological applications, which also helps in reducing environmental impacts and promote the development of an ecofriendly activity. Thus, nile tilapia viscera were subjected to proteolysis, then the glycosaminoglycans were complexed with ion exchange resin (Lewatit), it was fractionated with increasing volumes of acetone and purified by ion exchange chromatography DEAE-Sephacel. Further, the fraction was analyzed by agarose gel electrophoresis and nuclear magnetic resonance (NMR). The electrophoretic profile of the compound together the analysis of 1H NMR spectra and the HSQC correlation allow to affirm that the compound corresponds to a molecule like chondroitin sulfate. MTT assay was used to assess cell viability in the presence of CS tilapia isolated and showed that the compound is not cytotoxic to normal cells such as cells from the mouse embryo fibroblast (3T3). Then, this compound was tested for the ability to reduce the influx of leukocytes in model of acute peritonitis (in vivo) induced by sodium thioglycolate. In this context, it was done total and differential leukocytes counting in the blood and peritoneal fluid collected respectively from vena cava and the peritoneal cavity of the animals subjected to the experiment. The chondroitin sulfate for the first time isolated from tilapia (CST ) was able to reduce the migration of leukocytes to the peritoneal cavity of inflamed mice until 80.4 per cent at a dose 10µg/kg. The results also show that there was a significant reduction (p<0.001) of the population of polymorphonuclear leukocytes from peritoneal cavity in the three tested doses (0.1µg/kg; 1µg/kg and 10µg/kg) when it was compared to the positive control (just thioglycolate). Therefore, since the CST structure and mechanism of action has been completely elucidated, this compound may have potential for therapeutic use in inflammatory diseases

Avaliação anti-inflamatória e antipeçonhenta das espécies Hancornia speciosa e Mimosa tenuiflora em modelos experimentais de inflamação e envenenamento induzidos por Bothrops jararaca e Tityus serrulatus

Accidents caused by venomous animals represents a significant and serious public health problem in certain regions of Brazil, as well as in other parts of the world by the frequency with which they occur and the mortality they cause. The use of plant extracts as an antidote for poisoning cases is an ancient practice used in many communities that have no access to antivenom. Medicinal plants represent an important source of obtaining bioactive compounds able to assist directly in the treatment of poisoning or indirectly supplementing serum therapy currently used. The aim of this study was to evaluate the effect of extracts, fractions and isolated compounds from M. tenuiflora and H. speciosa in the inflammatory process induced by carrageenan and the venom of B. jararaca and T. serrulatus. The results showed that both M. tenuiflora and H. speciosa were capable of inhibiting cell migration and cytokines levels in peritonitis induced by carrageenin and venom of T. serrulatus. In poisoning by B. jararaca model, mice treated with the plants in studies decreased the leukocyte influx into the peritoneal cavity. Finally the M. tenuiflora and H. speciosa had antiphlogistic activity, reducing edema formation and exerted inhibitory action of leukocyte migration in local inflammation induced by the venom of B. jararaca. Through of Thin Layer Chromatography (TLC) analysis was possible identified the presence of flavonoids ,saponins and/or terpenes in aqueous extract of M. tenuiflora. By High Performance Liquid Chromatography analysis, it was possible to identify the presence of rutin and chlorogenic acid in aqueous extract of H. speciosa. We conclude that the administration of extracts, fractions and isolated compounds of H. speciosa and M. tenuiflora resulted in inhibition of the inflammatory process in different experimental models. This study demonstrates for the first time the effect of M. tenuiflora and H. speciosa in inhibition of the inflammation caused by B. jararaca and T. serrulatus venom.

Estratégias tróficas do peixe traíra, Hoplias malabaricus e do peixe cascudo, Hypostomus pusarum do açude Marechal Dutra, Rio Grande do Norte, Brasil

The fish, Hoplias malabaricus (Osteichthyes: Erythrinidae) and armored catfish, Hypostomus pusarum (Loricariidae) are of freshwater origin from the neotropical region, and are considered ecologically and economically of important. This work investigated the trophic strategies, the morphology and histology of the digestive tract of these fish captured from the Marechal Dutra reservoir, Acari, the semiarid region of Rio Grande do Norte, Brazil. A total of 133 individuals of H. malabaricus and 118 specimens of H. pusarum were analyzed. The two study species occupy different levels in the food chain and spatial distribution in the water column of the reservoir. The results of this study are presented in the form of four scientific papers. The first article describes the morphology and histology of the digestive tract and the feeding habits of H. malabaricus (Bloch, 1794). This fish has a short intestine, with an intestinal coefficient of 0.72 ± 0.09. The dietary importance index indicates that H. malabaricus feeds preferentially on animal matter, especially on fish (72.8%) and prawns (27.2%). The histology of its digestive tract confirms the carnivorous feeding habit. The second article discusses about the pioneering work on the feeding strategy and the characterization of anatomy and histology of the digestive tract of H. pusarum. The intestine of this fish is long, with an intestinal coefficient of 10.8±0.7. The dietary importance index indicates that H. pusarum feeds preferentially on organic matter in decomposition (88.7%) and on filamentous microalgae and diatoms (11.3%). The third article compares the morphological aspects of the digestive tract of H. malabaricus and H. pusarum, in relation to their food habits. The arrangement of the digestive organs in both species is directly related to the shape of the peritoneal cavity and the form of the body. The short intestine of H. malabaricus and the long intestine of H. pusarum are associated with their feeding habits. The morphology of the digestive tracts of H. malabaricus and H. pusarum confirm their food habits, carnivorous and detritivorous / herbivorous, respectively. The fourth article discusses the food and reproductive aspects (length and weight, length-weight relationship, type of growth and sex ratio) of H. pusarum. This species has a negatively allometric growth, with the predominance of females in the sampled population. H. malabaricus inhabits the pelagic environment and is a carnivore, while H. pusarum, lives in the benthic environment and is characterized as a detritivore/herbivore. Each species studied shows a very different diet, without trophic competition between them. The morphological and anatomical structures of the digestive tract reflect their feeding strategy.

Influência do tempo e dose de heparina em modelo de peritonite aguda

In the last years, heparin has become target of many studies related to inflammation due its ability of biding to proteins involved on immune response. Recently, it was demonstrated, at our laboratory, using a thIoglycollate-induced peritonitis model, heparin s capacity of reduce cellular influx into the peritoneal cavity, 3 hours after the inflammatory stimulus. Once neutrophilic infiltration is highest around 8 hours after the inflammatory stimulus, at the present work, using the same peritonitis model, it was assessed heparin s ability of keeping the interference on leukocyte infiltration, 8 hours after inflammation induction. Moreover, using cellular differential count, it was evaluated how the cellular populations involved in the inflammatory process would be affected by the treatment. Eight hours after the inflammatory stimulus, only heparin dosage of 1 μg/Kg was able to reduce the cellular influx to peritoneum, 62.8% of reduction when compared to positive control (p < 0.001). Furthermore, heparin dosage of 15 μg/Kg presented a pro-inflammatory effect in whole blood verified by the increase of 60.9% (p < 0.001) and 117.8% (p < 0.001) on neutrophils and monocytes proportion, respectively, when compared to positive control. In addition, this dosage also presented a neutrophilic proportion on peritoneal fluid 27.3% higher than positive control (p < 0.05). This duality between anti- and pro-inflammatory effects at different times corroborates studies that attribute a pleiotropic immunomodulator role to heparin.

Avaliação do potencial anti-inflamatório de composto tipo heparina (cCTH) extraído do caranguejo Goniopsis cruentata em modelo experimental de peritonite

Heparin, a sulfated polysaccharide, was the first compound used as an anticoagulant and antithrombotic agent. Due to their structural characteristics, also has great potential anti-inflammatory, though such use is limited in inflammation because of their marked effects on coagulation. The occurrence of heparin-like compounds that exhibit anticoagulant activity decreased in aquatic invertebrates, such as crab Goniopsis cruentata, sparked interest for the study of such compounds as anti-inflammatory drugs. Therefore, the objective of this study was to evaluate the potential modulator of heparin-like compound extracted from Goniopsis cruentata in inflammatory events, coagulation, and to evaluate some aspects of its structure. The heparin-type compound had a high degree of N-sulphation in its structure, being able to reduce leukocyte migration into the peritoneal cavity at lower doses compared to heparin and diclofenac sodium (anti-inflammatory commercial). Furthermore, it was also able to inhibit the production of nitric oxide and tumor necrosis factor alpha by activated macrophages, inhibited the activation of the enzyme neutrophil elastase in low concentrations and showed a lower anticoagulant effect in high doses as compared to porcine mucosal heparin. Because of these observations, the compound extracted from crab Goniopsis cruentata can be used as a structural model for future anti-inflammatory agents

Neurilemmoma of the oral cavity in pediatric patient

Neurilemmomas arising from Schwann cells, rarely found in oral cavity, are characterized by solitary occurrence, slow growth and smooth surface with variable clinical aspects, depending on the nerve origin. Most instances are typically asymptomatic. Microscopically, the histopathological characteristic consists of alternating regions of hypercellularity and hypocellularity such as Antoni A and Antoni B, respectively. The prognosis is favorable; complete surgical removal is the treatment of choice, no recurrence should be expected. The present report shows the case of neurilemmoma in bucal mucosa, in a 9-year-old child. Patient was referred to the Stomatology Clinic of School of Dentistry, Universidade Federal do Rio Grande do Norte, with complaint of nodular mass, with approximately one year development. Patient was submitted to excisional biopsy, with histopathological exam showing diagnosis of neurilemmoma. After 1 year follow-up there has been no recurrence of lesion.

Neurilemmoma of the oral cavity in pediatric patient

Neurilemmomas arising from Schwann cells, rarely found in oral cavity, are characterized by solitary occurrence, slow growth and smooth surface with variable clinical aspects, depending on the nerve origin. Most instances are typically asymptomatic. Microscopically, the histopathological characteristic consists of alternating regions of hypercellularity and hypocellularity such as Antoni A and Antoni B, respectively. The prognosis is favorable; complete surgical removal is the treatment of choice, no recurrence should be expected. The present report shows the case of neurilemmoma in bucal mucosa, in a 9-year-old child. Patient was referred to the Stomatology Clinic of School of Dentistry, Universidade Federal do Rio Grande do Norte, with complaint of nodular mass, with approximately one year development. Patient was submitted to excisional biopsy, with histopathological exam showing diagnosis of neurilemmoma. After 1 year follow-up there has been no recurrence of lesion.