POLISSACARÍDEOS DA CASCA DE Ximenia americana: ISOLAMENTO, CARACTERIZAÇÃO QUÍMICA E ATIVIDADES NA NOCICEPÇÃO E DOR PANCREÁTICA
Autor(es):
SILVA, KAIRA EMANUELLA SALES DA
Palavras Chaves:
Não informado
Ano de Publicação:
2014
Resumo:
POLISSACARÍDEOS DA CASCA DE Ximenia americana: ISOLAMENTO, CARACTERIZAÇÃO QUÍMICA E ATIVIDADES NA NOCICEPÇÃO E DOR PANCREÁTICA. Kaira Emanuella Sales da Silva. Orientadora: Profª Drª Maria Gonçalves Pereira. Curso de Mestrado Acadêmico em Ciências Fisiológicas, Universidade Estadual do Ceará, 2013. Polissacarídeos de vegetais têm se destacado por seus efeitos em várias patologias, contudo, os estudos na nocicepção e pancreatite são escassos. As cascas de Ximenia americana Linne são utilizada popularmente contra a dor gástrica e experimentalmente foi demonstrado a atividade antinociceptiva de extratos aquosos. Objetivou-se purificar e caracterizar os polissacarídeos da casca de X. americana e avaliar o seu efeito antinociceptivo em modelos clássicos e na pancreatite aguda induzida por ceruleína em camundongos. O pó seco da casca (5 g) foi despigmentado com metanol, filtrado e o material residual utilizado na extração dos polissacarídeos (NaOH 0,1 M; 3x). O sobrenadante da centrifugação foi neutralizado e os polissacarídeos totais (PLT) precipitados com Etanol. Os PLT foram dissolvidos em água destilada (2:1, p:v), aplicados em cromatografia de troca iônica (DEAE-celulose) e as frações polissacarídicas acídicas eluidas com NaCl (0,1-1 M). As frações majoritárias (FI e FII) foram dialisadas e liofilizadas. PLT e frações foram analisados quanto aos teores de carboidratos, ácido urônico e proteínas; ao grau de pureza, peso molecular e presença de ácido urônico (eletroforese em gel de agarose e poliacrilamida); e a composição monossacarídica (cromatografia gasosa acoplada a espectro de massa). Camundongos Swiss machos (20-30 g, CEUA/UECE 12.783.679-9) receberam os polissacarídeos (PLT e/ou FII) por administração intravenosa (i.v.; 0,1, 1 e 10 mg/kg) ou oral (v.o.; 100 mg/kg) e avaliados nos testes de nocicepção (formalina, contorções abdominais, von Frey e placa quente). A pancreatite aguda (PA) foi induzida por 10 injeções intraperitoneais de ceruleína (50 μg/kg, h/h) e os grupos experimentais receberam PLT (10 mg/kg, e.v.) em uma única dose, antes da indução, ou em duas, antes e após a indução da PA. Foram analisados os parâmetros: nível plasmático de amilase e lipase, atividade da mieloperoxidase-MPO pancreática e histologia do pâncreas. A hipernocicepção abdominal (teste de von Frey) foi avaliada na 11ª, 24ª e 36ª h. A coordenação motora (teste de rota-rod) e a toxicidade foram avaliadas 14 dias após tratamento dos animais com os PLT. Média ± SEM (n=6-8), ANOVA e teste de Bonferroni (p<0,05). Os polissacarídeos apresentaram elevados teores de carboidratos (PLT-47%), FI-39%, FII-44,5%) e ácido urônico (PLT-21%, FI-27,5%, FII-39%), e baixo teor proteico (PLT-6,5%, FI-2% FII-1,5%). A eletroforese em agarose revelou maior grau de purificação e presença de ácido urônico para FII, sendo esta composta predominantemente por arabinose, galactose e glicose. No teste da formalina os PLT inibiram em 48% (1 mg/kg) e 78% (10 mg/kg) a 1ª fase, e em 50% (0,1 mg/kg) e 93% (10 mg/kg) a 2ª fase do teste. Para FII este efeito foi de 41% (0,1 mg/kg) e 60% (1 mg/kg) apenas na segunda fase. No teste das contorções a inibição foi de 69% (10 mg/kg, e.v.) e 44% (100 mg/kg, v.o.) para os PLT e de 72% (0,1 mg/kg) e 58% (1 mg/kg) para FII. No teste de von Frey os PLT (10 mg/kg) aumentaram em 35% (2ª h) e em 50% (3ª h) o limiar de hipernocicepção e FII (1 mg/kg) aumentou em 69% na 1ª; 52,8% na 2ª e 63% na 3ª h. PLT e FII não mostraram efeito no teste da placa quente. Na PA os PLT (10 mg/kg) tanto em uma, como em duas doses aumentaram o limiar de hipernocicepção dos animais em 44% e 42%, respectivamente, já o tratamento com 2 doses diminuiu o processo inflamatório no pâncreas (MPO e histologia) e os níveis plasmáticos de amilase e lipase. Os PLT não alteraram a motricidade, a massa corpórea/órgãos e os marcadores hematológicos, de função renal e hepática dos animais. Os polissacarídeos de X. americana apresentam elevados teores de carboidratos e ácido urônico, sendo estes compostos principalmente por arabinose, glicose e galactose. Estes inibiram a nocicepção química e mecânica e reduziram a gravidade da pancreatite aguda, sem causar toxicidade nos animais. Palavras-chave: Ximenia americana; polissacarídeos; nocicepção; pancreatite 9 ABSTRACT POLYSACCHARIDE OF Ximenia americana barks: ISOLATION, CHARACTERIZATION AND ACTIVITY IN NOCICEPTION AND PANCREATIC PAIN-INDUCED BY CAERULEIN IN MICE. Supervisor: Profª Drª Maria Gonçalves Pereira. Curso de Mestrado Acadêmico em Ciências Fisiológicas, Universidade Estadual do Ceará, 2013. Plant polysaccharides have been highlighted for its effects in several pathologies, however, there are few studies in nociception and pancreatitis. Ximenia americana Linne (bark) is used to treat gastric pain and its analgesic effect had been demonstrated experimentally for the aqueous extract. This study aimed to purify and characterize the polysaccharides from X. americana barks and assess its antinociceptive effect and acute pancreatitis in mice. The barks powder (5 g) was depigmented with methanol, filtered and the residual material used in the extraction of polysaccharides (0.1 M NaOH; 3x). The supernatant of centrifugation was neutralized and the total polysaccharides (TPL) precipitated with Ethanol. TPL were dissolved in distilled water (2:1, w:v) and applied in ion exchange chromatography (DEAE-cellulose) and acidic polysaccharide fractions was eluted with NaCl (0.1-1 M). The majority polysaccharide fractions (FI and FII) were dialyzed and lyophilized. TPL and fractions were analyzed to assed the carbohydrate, uronic acid and protein contents; the purity, molecular weight and the presence of uronic acid (electrophoresis in agarose and polyacrylamide gels) and their monosaccharide composition (gas chromatography-mass spectrum). Male Swiss mice (20-30 g, CEUA/UECE 12.783.679-9) received the TPL or FII by intravenous (0.1, 1 and 10 mg/kg, i.v.) or oral (100 mg/kg, p.o.) routs and evaluated in models/tests of nociception (formalin, writhing, hot plate and von Frey). Acute pancreatitis (AP) was induced by 10 intraperitoneal injections of cerulein (50 μg/kg) and experimental groups received PLT (10 mg/kg, i.v.) in a single dose before induction, or two administrations, before and after induction of AP. After induction it was analyzed: plasma levels of amylase and lipase, activity of pancreatic myeloperoxidase-MPO and histology of the pancreas. Abdominal hypernociception was evaluated in 1st, 24th and 36th h. The motor coordination (rota-rod test) and subchronic toxicity (14 days) were evaluated after treatment of the animals with TPL. Mean ± SEM (n=6-8), ANOVA and Bonferroni's test (p<0.05). The polysaccharides showed high contents of total carbohydrates (TPL-47%, FI-39%; FII-44.5%), uronic acid (TPL-21%, FI-27.5%, FII-39%) and low protein content (TPL-6.5%, FI-2%, FII-1.5%). Agarose gel electrophoresis revealed a higher degree of purification for FII, which is composed predominantly of arabinose, galactose and glucose. TPL inhibited by 48% (1 mg/kg) and 78% (10 mg/kg) the 1st and by 50% (0.1 mg/kg) and 93% (10 mg/kg) the 2nd phase of formalin. For FII this effect was 41% (0.1 mg/kg) and 60% (1 mg/kg) only in the second phase of the test. In writhing test, the inhibition was by 69% (10 mg/kg, i.v.), 44% (100 mg/kg, p.o.) for PLT, 72% (0.1 mg/kg), and 58% (1 mg/kg) for FII. In von Frey test, TPL (10 mg/kg) increased by 35% (2nd h) and 50% (3rd h) the hypernociception threshold and FII (1 mg/kg) increased by 69% in 1st; 52.8% in 2nd and 63% in 3rd h. TPL and FII showed no effect in the hot plate test. TPL (10 mg/kg, 1 or 2 doses) increased the threshold by 44% and 42% in abdominal hypernociception, respectively. TPL (two doses) inhibited the inflammatory process in pancreas (MPO and histology) and decreased serum levels of amylase and lipase. TPL did not alter animals motricity, body/organs mass and hematological marker (renal and hepatic). The polysaccharides of X. americana show high levels of carbohydrates and uronic acid, especially arabinose, galactose and glucose. These polysaccharides inhibit chemical and mechanical nociception and reduced the severity of acute pancreatitis, without affecting motor animals coordination and induce toxicity. Keywords: Ximenia
Abstract:
POLYSACCHARIDE OF Ximenia americana barks: ISOLATION, CHARACTERIZATION AND ACTIVITY IN NOCICEPTION AND PANCREATIC PAIN-INDUCED BY CAERULEIN IN MICE. Supervisor: Profª Drª Maria Gonçalves Pereira. Curso de Mestrado Acadêmico em Ciências Fisiológicas, Universidade Estadual do Ceará, 2013. Plant polysaccharides have been highlighted for its effects in several pathologies, however, there are few studies in nociception and pancreatitis. Ximenia americana Linne (bark) is used to treat gastric pain and its analgesic effect had been demonstrated experimentally for the aqueous extract. This study aimed to purify and characterize the polysaccharides from X. americana barks and assess its antinociceptive effect and acute pancreatitis in mice. The barks powder (5 g) was depigmented with methanol, filtered and the residual material used in the extraction of polysaccharides (0.1 M NaOH; 3x). The supernatant of centrifugation was neutralized and the total polysaccharides (TPL) precipitated with Ethanol. TPL were dissolved in distilled water (2:1, w:v) and applied in ion exchange chromatography (DEAE-cellulose) and acidic polysaccharide fractions was eluted with NaCl (0.1-1 M). The majority polysaccharide fractions (FI and FII) were dialyzed and lyophilized. TPL and fractions were analyzed to assed the carbohydrate, uronic acid and protein contents; the purity, molecular weight and the presence of uronic acid (electrophoresis in agarose and polyacrylamide gels) and their monosaccharide composition (gas chromatography-mass spectrum). Male Swiss mice (20-30 g, CEUA/UECE 12.783.679-9) received the TPL or FII by intravenous (0.1, 1 and 10 mg/kg, i.v.) or oral (100 mg/kg, p.o.) routs and evaluated in models/tests of nociception (formalin, writhing, hot plate and von Frey). Acute pancreatitis (AP) was induced by 10 intraperitoneal injections of cerulein (50 μg/kg) and experimental groups received PLT (10 mg/kg, i.v.) in a single dose before induction, or two administrations, before and after induction of AP. After induction it was analyzed: plasma levels of amylase and lipase, activity of pancreatic myeloperoxidase-MPO and histology of the pancreas. Abdominal hypernociception was evaluated in 1st, 24th and 36th h. The motor coordination (rota-rod test) and subchronic toxicity (14 days) were evaluated after treatment of the animals with TPL. Mean ± SEM (n=6-8), ANOVA and Bonferroni's test (p<0.05). The polysaccharides showed high contents of total carbohydrates (TPL-47%, FI-39%; FII-44.5%), uronic acid (TPL-21%, FI-27.5%, FII-39%) and low protein content (TPL-6.5%, FI-2%, FII-1.5%). Agarose gel electrophoresis revealed a higher degree of purification for FII, which is composed predominantly of arabinose, galactose and glucose. TPL inhibited by 48% (1 mg/kg) and 78% (10 mg/kg) the 1st and by 50% (0.1 mg/kg) and 93% (10 mg/kg) the 2nd phase of formalin. For FII this effect was 41% (0.1 mg/kg) and 60% (1 mg/kg) only in the second phase of the test. In writhing test, the inhibition was by 69% (10 mg/kg, i.v.), 44% (100 mg/kg, p.o.) for PLT, 72% (0.1 mg/kg), and 58% (1 mg/kg) for FII. In von Frey test, TPL (10 mg/kg) increased by 35% (2nd h) and 50% (3rd h) the hypernociception threshold and FII (1 mg/kg) increased by 69% in 1st; 52.8% in 2nd and 63% in 3rd h. TPL and FII showed no effect in the hot plate test. TPL (10 mg/kg, 1 or 2 doses) increased the threshold by 44% and 42% in abdominal hypernociception, respectively. TPL (two doses) inhibited the inflammatory process in pancreas (MPO and histology) and decreased serum levels of amylase and lipase. TPL did not alter animals motricity, body/organs mass and hematological marker (renal and hepatic). The polysaccharides of X. americana show high levels of carbohydrates and uronic acid, especially arabinose, galactose and glucose. These polysaccharides inhibit chemical and mechanical nociception and reduced the severity of acute pancreatitis, without affecting motor animals coordination and induce toxicity. Keywords: Ximenia americana; polysaccharides; nociception; pancreatitis 10
Tipo do Trabalho:
Dissertação
Referência:
SILVA, KAIRA EMANUELLA SALES DA. POLISSACARÍDEOS DA CASCA DE Ximenia americana: ISOLAMENTO, CARACTERIZAÇÃO QUÍMICA E ATIVIDADES NA NOCICEPÇÃO E DOR PANCREÁTICA. 2014. 97 f. Dissertação (Mestrado Acadêmico ou Profissional em 2014) - Universidade Estadual do Ceará, , 2014. Disponível em: Acesso em: 28 de abril de 2024
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