RESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Geoffroea decorticans (chañar) fruits and their derivate product (arrope) have been traditionally used as food and a folk medicine for the treatment of a wide variety of diseases including bronchopulmonary disorders and to relieve dolorous process. AIM OF THE STUDY: In order to evaluate the pharmacology action of this plant, studies were performed of antinociceptive and antioxidant activities. MATERIALS AND METHODS: The aqueous and ethanolic extracts and arrope of chañar were evaluated in various established pain models, including chemical nociception induced by subplantar formalin and intraperitoneal acetic acid and thermal nociception method, such as tail immersion test in rats. To examine the possible connection of the opioid receptor to the antinociceptive activity of extracts and arrope it was performed a combination test with naloxone, a non-selective opioid receptor antagonist. RESULTS: The aqueous extract and arrope (1000 mg/kg) caused an inhibition of the pain in formalin test in the first phase, similar to morphine and decrease in the second phase. In a combination test using naloxone, diminished analgesic activity of aqueous extract and arrope were observed, indicating that antinociceptive activity is connected with the opioid receptor. The aqueous extract and arrope, caused an inhibition of the writhing response induced by acetic acid. Central involvement in analgesic profile was confirmed by the tail immersion test, in which the aqueous extract and arrope showed a significant analgesic activity by increasing latency time. The aqueous extract showed higher antioxidant activity than the arrope, it may be due to the cooking process. CONCLUSIONS: This study has shown that the aqueous extract and arrope of Geoffroea decorticans (chañar) fruits, does possess significant antinociceptive effects. It is further concluded that aqueous extract with maximum inhibition of free radical is the most potent extract amount tested extracts. At the oral doses tested the aqueous extract and arrope were non-toxic. The present results justifies their popular use and constitutes the first validation study of the antinociceptive action.
Assuntos
Analgésicos/uso terapêutico , Fabaceae/química , Frutas/química , Interações Ervas-Drogas , Naloxona/farmacologia , Dor/tratamento farmacológico , Fitoterapia/métodos , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Analgésicos/farmacologia , Animais , Antioxidantes/farmacologia , Modelos Animais de Doenças , Flavonoides/análise , Masculino , Antagonistas de Entorpecentes , Dor/induzido quimicamente , Medição da Dor/métodos , Soluções Farmacêuticas/administração & dosagem , Soluções Farmacêuticas/farmacologia , Soluções Farmacêuticas/uso terapêutico , Fenóis/análise , Extratos Vegetais/administração & dosagem , Extratos Vegetais/antagonistas & inibidores , Ratos , Ratos WistarRESUMO
Amphibians respond to microbial infection through cellular and humoral defense mechanisms such as antimicrobial protein secretion. Most humoral defense proteins are synthetized in the skin. In this study we isolated two beta-galactoside-binding lectins with molecular weights of 50 and 56 KDa from the skin of Bufo arenarum. These lectins have significant hemagglutination activity against trypsinized rabbit erythrocytes, which was inhibited by galactose-containing saccharides. They are water-soluble and independent of the presence of calcium. The antimicrobial analysis for each lectin was performed. At mumolar concentration lectins show strong bacteriostatic activity against Gram negative bacteria (Escherichia coli K12 4100 and wild strains of Escherichia coli and Proteus morganii) and Gram positive bacteria (Enterococcus faecalis). The antibacterial activity of these lectins may provide an effective defense against invading microbes in the amphibian Bufo arenarum. (AU)
Assuntos
Estudo Comparativo , RESEARCH SUPPORT, NON-U.S. GOVT , Antibacterianos/farmacologia , Bufo arenarum/metabolismo , Lectinas/farmacologia , Pele/química , Bufo arenarum/anatomia & histologia , Enterococcus faecalis/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Testes de Hemaglutinação , Hemaglutininas/metabolismo , Lactose/metabolismo , Lectinas/metabolismo , Proteus/efeitos dos fármacos , CoelhosRESUMO
Amphibians respond to microbial infection through cellular and humoral defense mechanisms such as antimicrobial protein secretion. Most humoral defense proteins are synthetized in the skin. In this study we isolated two beta-galactoside-binding lectins with molecular weights of 50 and 56 KDa from the skin of Bufo arenarum. These lectins have significant hemagglutination activity against trypsinized rabbit erythrocytes, which was inhibited by galactose-containing saccharides. They are water-soluble and independent of the presence of calcium. The antimicrobial analysis for each lectin was performed. At mumolar concentration lectins show strong bacteriostatic activity against Gram negative bacteria (Escherichia coli K12 4100 and wild strains of Escherichia coli and Proteus morganii) and Gram positive bacteria (Enterococcus faecalis). The antibacterial activity of these lectins may provide an effective defense against invading microbes in the amphibian Bufo arenarum.
Assuntos
Antibacterianos/farmacologia , Bufo arenarum/metabolismo , Lectinas/farmacologia , Pele/química , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Bufo arenarum/anatomia & histologia , Enterococcus faecalis/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Testes de Hemaglutinação , Hemaglutininas/metabolismo , Lactose/metabolismo , Lectinas/metabolismo , Proteus/efeitos dos fármacos , CoelhosRESUMO
Although the nephrotoxic effects of lead are well documented, the subcellular mechanisms of its action on the kidney remain unclear. The aim of the present work was to investigate the effects of chronic lead exposure on the expression of laminin-1 and fibronectin in the kidney of lead-treated rats. Western immunoblotting of the kidney extracts revealed that experimental exposure to lead resulted in a marked decrease in the intensity of the bands corresponding to laminin-1 and an increase in the intensity of the band corresponding to fibronectin. Immunohistochemical studies demonstrated a weak labelling to laminin-1 and a strong labelling to fibronectin in all renal basement membranes together with a decrease in their thickness. Other ultrastructural alterations found were a diminution in the amount of endothelial fenestrae, an increased fusion of foot processes in epithelial cells of the glomerulus and the presence of intranuclear inclusion bodies in the proximal tubule cells. Lead intoxication might be responsible for the above alterations in the renal extracellular matrix that could play an important role in the pathogenesis of lead nephropathy.
Assuntos
Matriz Extracelular/efeitos dos fármacos , Rim/efeitos dos fármacos , Chumbo/toxicidade , Animais , Membrana Basal/efeitos dos fármacos , Membrana Basal/ultraestrutura , Western Blotting , Modelos Animais de Doenças , Eletroforese em Gel de Poliacrilamida , Matriz Extracelular/ultraestrutura , Fibronectinas/biossíntese , Técnicas Imunoenzimáticas , Rim/metabolismo , Glomérulos Renais/efeitos dos fármacos , Glomérulos Renais/ultraestrutura , Laminina/biossíntese , Chumbo/sangue , Intoxicação por Chumbo , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
The hypoglycemic effect of the water extract of the leaves of Smallantus sonchifolius (yacon) was examined in normal, transiently hyperglycemic and streptozotocin (STZ)-induced diabetic rats. Ten-percent yacon decoction produced a significant decrease in plasma glucose levels in normal rats when administered by intraperitoneal injection or gastric tube. In a glucose tolerance test, a single administration of 10% yacon decoction lowered the plasma glucose levels in normal rats. In contrast, a single oral or intraperitoneal administration of yacon decoction produced no effect on the plasma glucose levels of STZ-induced diabetic rats. However, the administration of 2% yacon tea ad libitum instead of water for 30 days produced a significant hypoglycemic effect on STZ-induced diabetic rats. After 30 days of tea administration, diabetic rats showed improved body (plasma glucose, plasma insulin levels, body weight) and renal parameters (kidney weight, kidney to body weight ratio, creatinine clearance, urinary albumin excretion) in comparison with the diabetic controls. Our results suggest that yacon water extract produces an increase in plasma insulin concentration.
Assuntos
Diabetes Mellitus Experimental/tratamento farmacológico , Hipoglicemiantes/farmacologia , Plantas Medicinais/química , Albuminúria/metabolismo , Animais , Argentina , Glicemia/metabolismo , Creatinina/metabolismo , Diabetes Mellitus Experimental/sangue , Teste de Tolerância a Glucose , Insulina/sangue , Masculino , Extratos Vegetais/farmacologia , Folhas de Planta/química , Ratos , Ratos Sprague-DawleyRESUMO
Diabetes mellitus is characterized by anatomical and functional alterations of the intestinal tract. However, the aetiology of these disturbances remains unclear. The aim of the present work was to investigate the effects of diabetes on the expression of laminin-1 and fibronectin in the small intestine of Streptozotocin (STZ)-induced diabetic rats. The Western immunoblotting of the extracts from the small intestine revealed that experimental diabetes resulted in a marked increase in the intensity of the bands corresponding to laminin-1 and fibronectin. Immunohistochemical studies demonstrated a strong labelling to these two extracellular matrix (ECM) proteins in the small intestine of diabetic rats, mainly localized in the smooth muscle layer. These results occur together with a thickening of the basement membrane (BM) of the smooth muscle cells, demonstrated by transmission electron microscopy (TEM). We propose that the accumulation of ECM proteins in the smooth muscle layer may be an effect mediated by hyperglycaemia, since insulin treatment of diabetic rats reversed this accumulation. These results could provide information on the potential role of the ECM in the intestine, an organ which is known to exhibit important alterations in diabetes.
Assuntos
Diabetes Mellitus Experimental/metabolismo , Proteínas da Matriz Extracelular/metabolismo , Intestino Delgado/metabolismo , Músculo Liso/metabolismo , Animais , Membrana Basal/metabolismo , Membrana Basal/patologia , Membrana Basal/ultraestrutura , Glicemia/metabolismo , Western Blotting , Peso Corporal , Diabetes Mellitus Experimental/patologia , Fibronectinas/metabolismo , Intestino Delgado/patologia , Intestino Delgado/ultraestrutura , Laminina/metabolismo , Masculino , Músculo Liso/patologia , Músculo Liso/ultraestrutura , Tamanho do Órgão , Ratos , Ratos Sprague-DawleyRESUMO
Diabetes mellitus is associated with various structural and functional liver abnormalities that affect the glycogen and lipid metabolisms. The effects of streptozotocin-induced diabetes and of insulin supplementation to Sprague-Dawley diabetic rats on ganglioside patterns in liver were determined. Diabetic livers showed a tendency to hepatomegaly 3 weeks after STZ-induction of diabetes. The concentration of total gangliosides in diabetic and non-diabetic livers was similar, but the concentration of total gangliosides in the liver of insulin-stabilized rats was slightly increased. Bidimensional TLC chromatographic analysis of gangliosides isolated from normal diabetic and insulin-stabilized diabetic livers showed quantitative and qualitative changes. In comparison with normal controls, the densitometric analyses of diabetic liver ganglioside patterns had increased amounts of GM3, GM1, GD1b, and GT1b gangliosides, while GM2 could not be detected. The hepatic ganglioside pattern of insulin-stabilized diabetic rats was partially restored, resembling the profile of normal rats. The activity of GalNAcT, GalT-2 and SialT-4 transferases was measured in liver microsomal fractions of the different groups of animals. Diabetic rats showed an increased activity of GalNAcT and a decrease in the activity of GalT-2 and SialT-4 compared with the controls. The enzymatic activities found in insulin-treated rats showed a tendency to return to the values observed in normal control animals. The results evidenced that streptozotocin-induced diabetes affects the liver ganglioside pattern and the ganglioside synthesis enzyme activity. The alterations found in ganglioside metabolism could represent one of the earliest changes associated with the diabetic pathology.
Assuntos
Diabetes Mellitus Experimental/metabolismo , Gangliosídeos/metabolismo , Glicoesfingolipídeos/metabolismo , Fígado/metabolismo , Animais , Glicemia/metabolismo , Diabetes Mellitus Experimental/tratamento farmacológico , Gangliosídeos/isolamento & purificação , Glicoesfingolipídeos/isolamento & purificação , Glicosiltransferases/metabolismo , Insulina/uso terapêutico , Fígado/efeitos dos fármacos , Fígado/patologia , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
The aim of the present study was to determine the presence of the connexins Cx43, Cx32 and Cx26 in Bufo arenarum ovarian follicles during the breeding season as well as to analyse the possible alterations in the meiotic process when connexins are blocked by specific antibodies. Western blot analysis revealed that the Cx43 and Cx32 proteins were present but not Cx26. We demonstrated that the anti-Cx43 and anti-Cx32 antibodies produced the uncoupling of the gap junctions. When these junctions are blocked the maturation process is triggered in the oocytes. We determined that dbcAMP exerts an inhibitory effect on the maturation induced by the uncoupling of the gap junctions when the oocytes are injected or pretreated with this metabolite. We propose the idea that cAMP is the regulatory molecule in meiotic arrest in this amphibian species.
Assuntos
Bufo arenarum/fisiologia , AMP Cíclico/metabolismo , Junções Comunicantes/fisiologia , Meiose , Folículo Ovariano/fisiologia , Animais , Anticorpos Monoclonais/farmacologia , Western Blotting , Bucladesina/farmacologia , Conexina 26 , Conexina 43/imunologia , Conexina 43/metabolismo , Conexinas/imunologia , Conexinas/metabolismo , Feminino , Junções Comunicantes/efeitos dos fármacos , Folículo Ovariano/citologia , Progesterona/farmacologia , Proteína beta-1 de Junções ComunicantesRESUMO
In the present paper we established the ganglioside composition of the blastula and gastrula stages of the anuran amphibian Bufo arenarum, two relevant stages characterized by dynamic changes in morphology and cellular rearrangements. Densitometric studies evidenced that GD1a and GT1b were the more abundant gangliosides of the blastula embryos whereas GM1 and GM2 were the predominant species in gastrula embryos. Analysis of ganglioside abundance indicates that the "a" and "b" synthesis pathways perform similar biosynthetic activities in the blastula stage, in contrast to the gastrula stage in which a marked predominance of the "a" pathway occurred. The spatio-temporal expression of GM1 and of polygangliotetraosyl ceramides (pGTC) was investigated by wholemount immunocytochemistry using cholera toxin B subunit (CTB) and an affinity purified human anti-GM1 antibody. The pGTC were detected as GM1 after treatment with neuraminidase. Blastomeres from the inner surface of the blastocoelic roof (BCR) of blastula embryos were GM1 and pGTC positive. At midgastrula stage, embryos showed an increased labeling on the inner surface of BCR. To establish whether the GM1 ganglioside was involved in the gastrulation processes, CTB, anti-GM1 antibodies and anti-GM1 Fab' fragments were microinjected into the blastocoel cavity of blastula embryos. Treatment with the probes blocked gastrulation. Scanning electron microscopy analysis of blocked embryos revealed that mesodermal cell migration, radial interdigitation, and convergent extension movements were affected. The blocking of gastrulation was correlated with the absence of fibronectin and EP3/EP4 on the inner surface of blastocoelic roof of CTB- or anti-GM1 treated embryos. Results show that the GM1 ganglioside is differentially expressed by embryonic cells and participates in the morphogenetic processes of amphibian gastrulation. J. Exp. Zool. 286:457-472, 2000.
Assuntos
Bufo arenarum/embriologia , Gangliosídeo G(M1)/metabolismo , Gástrula/fisiologia , Animais , Bufo arenarum/fisiologia , Cromatografia em Camada Fina , Densitometria , Eletroforese em Gel de Poliacrilamida , Proteínas da Matriz Extracelular/metabolismo , Gangliosídeos/análise , Humanos , MicroinjeçõesRESUMO
The present study analyses, by transmission electron microscopy, vitellogenesis in two anuran amphibian families: Leptodactilidae (Ceratophrys cranwelli) and Bufonidae (Bufo arenarum). These differ in the type of stimulus that sets off their reproductive period, pluvial changes being the trigger in C. cranwelli and temperature increase in B. arenarum. We found that vitellogenesis follows an endocytic pathway that involves membranous structures (coated pits, coated vesicles, endosomes and multivesicular bodies). This process results in a fully grown yolk platelet of similar structure in both species. Despite the above similarity, a distinctive feature in B. arenarum was that the multivesicular bodies exhibited condensed proteins together with lipid droplets, the latter remaining as such even in the primordial yolk platelet. In C. cranwelli, however, lipids droplets were only found attached to the primordial yolk platelet. The coexistence of lipid droplets together with proteins in the nascent precursor yolk platelets observed in B. arenarum is similar to that found in B. marinus. This fact might constitute a characteristic feature of the Bufonidae family.