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Phytomodulatory proteins from the latex of the medicinal plant Calotropis procera has been shown to be implicated in many pharmacological properties. However there is no current ...information about their activity on glucose metabolism, although the latex is used in folk medicine for treating diabetes. In this study the phytomodulatory proteins (LP) from C. procera latex were assessed on glycemic homeostasis.
Control animals received a single intravenous dose (5 mg/kg) of LP or saline solution (CTL). Four hours after treatment, the animals were euthanized and their livers were excised for analysis by western blot and RT-PCR AMP-activated protein kinase (AMPK), phosphoenolpyruvate carboxykinase (PEPCK) and tumor necrosis factor alpha (TNF-α). In vivo tests of intraperitoneal tolerance to insulin, glucose and pyruvate were also performed, and the effect of LP administration on fed glycemia was studied followed by blood analysis to determine serum insulin levels.
Treatment with LP reduced glycemia two hours after glucose administration (LP: 87.2 ± 3.70 mg/dL versus CTL: 115.6 ± 8.73 mg/dL). However, there was no change in insulin secretion (CTL: 14.16 ± 0.68 mUI/mL and LP: 14.96 ± 0.55 mUI/mL). LP improved the insulin sensitivity, represented by a superior glucose decay constant during an insulin tolerance test (kITT) (4.17 ± 0.94%/min) compared to the CTL group (0.82 ± 0.72%/min), and also improved glucose tolerance at 30 min (105.2 ± 12.4 mg/dL versus 154.2 ± 18.51 mg/dL), while it decreased hepatic glucose production at 15 and 30 min (LP: 75.5 ± 9.31 and 52.5 ± 12.05 mg/dL compared to the CTL: 79.0 ± 3.02 and 84.5 ± 7.49 mg/dL). Furthermore, there was a significant inhibition of gene expression of PEPCK (LP: 0.66 ± 0.06 UA and CTL: 1.14 ± 0.22 UA) and an increase of phosphorylated AMPK (LP: 1.342 ± 0.21 UA versus CTL: 0.402 ± 0.09 UA). These findings confirm the effect of LP on glycemic control and suggest LP may be useful in diabetes treatment. However, the pharmacological mechanism of LP in PEPCK modulation still needs more clarification.
This study aimed at investigating the structural properties and mechanisms of the antifungal action of CpOsm, a purified osmotin from Calotropis procera latex. Fluorescence and CD assays revealed ...that the CpOsm structure is highly stable, regardless of pH levels. Accordingly, CpOsm inhibited the spore germination of Fusarium solani in all pH ranges tested. The content of the secondary structure of CpOsm was estimated as follows: α-helix (20%), β-sheet (33%), turned (19%) and unordered (28%), RMSD 1%. CpOsm was stable at up to 75°C, and thermal denaturation (Tm) was calculated to be 77.8°C. This osmotin interacted with the negatively charged large unilamellar vesicles (LUVs) of 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-rac-1-glycerol (POPG), inducing vesicle permeabilization by the leakage of calcein. CpOsm induced the membrane permeabilization of spores and hyphae from Fusarium solani, allowing for propidium iodide uptake. These results show that CpOsm is a stable protein, and its antifungal activity involves membrane permeabilization, as property reported earlier for other osmotins and thaumatin-like proteins.
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►In this study we reported new insights into structure and antifungal properties of a latex protein. ►The protein was highly stable and active regardless pH conditions. ►It interacted and induced permeabilization of negatively charged large unilamellar vesicles. ►It also induced the membrane permeabilization of spores and hyphae from Fusarium solani, allowing for propidium iodide uptake.
A Disbiose Intestinal é um estado no qual a microbiota gera efeitos prejudiciais, causadas por meio de mudanças qualitativas e quantitativas na própria microbiota. A Disbiose é um distúrbio cada vez ...mais relevante, podendo agir como causa ou coadjuvante no desenvolvimento de Doenças Crônicas Não Transmissíveis. O objetivo deste estudo é avaliar a prevalência de Disbiose intestinal através da detecção de sinais e sintomas na população de acadêmicos do curso de Nutrição de uma Instituição de Ensino Superior de Fortaleza – CE e identificar as Doenças Crônicas Não Transmissíveis mais associadas. Trata-se de um estudo transversal, com dados coletados em 2016, em uma amostra de 91 pessoas. A coleta de dados foi realizada através da aplicação de dois questionários: um questionário elaborado sobre Doenças Crônicas Não Transmissíveis e o Questionário de Rastreamento Metabólico do Institute for Functional Medicine. Os resultados sugerem que é alta a prevalência de sinais e sintomas de Disbiose Intestinal entre os acadêmicos de Nutrição (53,84%). Dentre os 33 participantes que apresentaram >40 pontos no Questionário de Rastreamento Metabólico, 45,45% (n=15) apresentaram Doenças Crônicas Não Transmissíveis e fatores de risco associados, 30,30% (n=10) apresentaram somente Doenças Crônicas Não Transmissíveis, 21,21% (n=7) apresentaram somente fatores de risco associados e apenas 3,03% (n=1) não teve nenhum critério associado. Portanto, pode-se observar que a prevalência de Disbiose entre os estudantes de Nutrição é altíssima e este distúrbio esteve prevalentemente associado a presença de Doenças Crônicas Não Transmissíveis e/ou com o seu possível desenvolvimento. ABSTRACT Prevalence of intestinal disbiosis and its relationship with chronic diseases not transmissible in students of a higher education institution of Fortaleza-CE The aim of this study is to evaluate the prevalence of intestinal Dysbiosis through detection of signs and symptoms in population of academics of the nutrition course of an institution of higher education in Fortaleza-CE and identify the chronic non-communicable diseases associated. This is a cross-sectional study, with data collected in 2016, in a sample of 91 people. The data were collected through the application of two questionnaires: a questionnaire elaborated on chronic non-communicable diseases and metabolic trace Survey Institute for Functional Medicine. The results suggest that the high prevalence of signs and symptoms of Intestinal Dysbiosis between academics of nutrition (53.84%). Among the 33 participants who presented > 40 points in the questionnaire of Metabolic Trace, 45.45% (n = 15) showed non-communicable chronic diseases and associated risk factors, 30.30% (n = 10) showed only non-communicable chronic diseases, 21.21% (n = 7) showed only risk factors and only 3.03% (n = 1) had no associated criteria. Therefore, we can see that the prevalence of Dysbiosis among students of nutrition is very high and this disorder was predominantly associated with the presence of chronic non-communicable diseases and/or with your possible development.
An osmotin (CpOsm) from the latex of Calotropis procera has been crystallized in both tetragonal and trigonal forms suitable for structure determination. Crystallographic studies of CpOsm are of ...great interest because limited information is available concerning the structure of latex proteins and CpOsm has previously been shown to interact with the spore membranes of some plant pathogenic fungi, thus impairing spore germination and hyphal growth. CpOsm crystals were grown using 0.1 M HEPES buffer pH 7.5, 26% PEG 4000, 0.2 M ammonium sulfate (space group P43) or using 0.1 M HEPES buffer pH 7.5, 35% MPD, 0.7 M ammonium sulfate (space group P3112). X‐ray diffraction data were collected to 2.17 Å (P43) and 1.80 Å (P3112) resolution and molecular‐replacement analyses produced initial phases for both crystal forms.
The aim of this study is to evaluate the prevalence of intestinal Dysbiosis through detection of signs and symptoms in population of academics of the nutrition course of an institution of higher ...education in Fortaleza-CE and identify the chronic non-communicable diseases associated. This is a cross-sectional study, with data collected in 2016, in a sample of 91 people. The data were collected through the application of two questionnaires: a questionnaire elaborated on chronic non-communicable diseases and metabolic trace Survey Institute for Functional Medicine. The results suggest that the high prevalence of signs and symptoms of Intestinal Dysbiosis between academics of nutrition (53.84%). Among the 33 participants who presented > 40 points in the questionnaire of Metabolic Trace, 45.45% (n = 15) showed non-communicable chronic diseases and associated risk factors, 30.30% (n = 10) showed only noncommunicable chronic diseases, 21.21% (n = 7) showed only risk factors and only 3.03% (n = 1) had no associated criteria. Therefore, we can see that the prevalence of Dysbiosis among students of nutrition is very high and this disorder was predominantly associated with the presence of chronic noncommunicable diseases and/or with your possible development. Key word: Dysbiosis. Chronic diseases. Risk factors. A Disbiose Intestinal e um estado no qual a microbiota gera efeitos prejudiciais, causadas por meio de mudancas qualitativas e quantitativas na propria microbiota. A Disbiose e um disturbio cada vez mais relevante, podendo agir como causa ou coadjuvante no desenvolvimento de Doencas Cronicas Nao Transmissiveis. O objetivo deste estudo e avaliar a prevalencia de Disbiose intestinal atraves da deteccao de sinais e sintomas na populacao de academicos do curso de Nutricao de uma Instituicao de Ensino Superior de Fortaleza--CE e identificar as Doencas Cronicas Nao Transmissiveis mais associadas. Trata-se de um estudo transversal, com dados coletados em 2016, em uma amostra de 91 pessoas. A coleta de dados foi realizada atraves da aplicacao de dois questionarios: um questionario elaborado sobre Doencas Cronicas Nao Transmissiveis e o Questionario de Rastreamento Metabolico do Institute for Functional Medicine. Os resultados sugerem que e alta a prevalencia de sinais e sintomas de Disbiose Intestinal entre os academicos de Nutricao (53,84%). Dentre os 33 participantes que apresentaram >40 pontos no Questionario de Rastreamento Metabolico, 45,45% (n=15) apresentaram Doencas Cronicas Nao Transmissiveis e fatores de risco associados, 30,30% (n=10) apresentaram somente Doencas Cronicas Nao Transmissiveis, 21,21% (n=7) apresentaram somente fatores de risco associados e apenas 3,03% (n=1) nao teve nenhum criterio associado. Portanto, pode-se observar que a prevalencia de Disbiose entre os estudantes de Nutricao e altissima e este disturbio esteve prevalentemente associado a presenca de Doencas Cronicas Nao Transmissiveis e/ou com o seu possivel desenvolvimento. Palavra-chave: Disbiose. Doencas Cronicas. Fatores de Risco.
OLIVEIRA, Raquel Sombra Basílio de. Caracterização bioquímica e mecanismo de ação do efeito protetor in vivo de proteínas do látex de Calotropis procera (Ait.) R. Br. sobre infecção letal induzida ...por Salmonella enterica subespécie enterica sorotipo Typhimurium. 2010. 109 f. Dissertação (Mestrado em Bioquímica) - Universidade Federal do Ceará, Fortaleza-CE, 2010.
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Previous issue date: 2010
Latex of the medicinal plant Calotropis procera (Apocynaceae) possesses molecules displaying different pharmacological activities, including pro- and anti-inflammatory. In this study immune inflammatory modulation of the soluble protein fraction (LP) recovered from the latex was investigated in experimental models of sepsis induced by CLP and inoculation of the S. Typhimurium. Biochemical aspects of laticifers proteins were investigated and biochemical, pharmacological and histopathological parameters were evaluated in experimental animals. LP protected animals and a unique dose (30 mg/Kg; i.p.) lead to 100% survival while non-treated infected animals (Salmonella group) reached 100% mortality within 24 h. Protection was only observed when LP was given 24 earlier of infection. LP did not exhibit in vitro bactericide activity suggesting indirect mechanism of action, probably by immune modulation. After 4 and 24 h of infection bacteria was similarly disseminated in liver, spleen and peritoneal fluid, local of bacteria inoculum. However, in blood viable bacteria was reduced only in animals given laticifers proteins. The protective effect of LP was also observed in its three sub-fraction, denominated of PI, PII and PIII, obtained after protein fractionation by ion exchange chromatography on a CM-Sepharose performed at pH 5.0. Neither, heat-treatment (100 C, 30 min) nor inhibition of its endogenous proteolytic enzymes, by iodoacetamide, eliminated the protective effect of LP. Nitric oxide (NO) an important signaling molecule was augmented in serum of non-treated animals whereas it was unaltered in control and LP/PI-treated animals 24 h after infection. Increasing of NO in serum is known to directly contribute to inhibit neutrophil migration in septic animals. Accordingly, failure of neutrophil migration to the infectious focus in septic animals was confirmed. Conversely, in animals given LP and PI, influx of neutrophils was evident. In addition, NO was also elevated in the infectious focus of LP/PI treated animals, probably due neutrophil activity as part of their microbicidal activity. Activity of adenosine deaminase (ADA) was measured locally after 4 and 24 h of infection was initiated. ADA was augmented in LP/PI treated animals and unaltered in non treated animals. Thrombocytopenia was observed in non treated animals but not in LP/PI protected animals. Neutrophilia and lymphopenia were documented in non-treated animals. Neutrophilia would result of high NO content in serum, which inhibits neutrophil migration and lymphopenia is part of immune suppression caused by sepsis. Neutrophilia was observed 7 days after infection in animals given LP/PI, suggesting a hematopoiesis stimulus. Lymphopenia first observed (24 h) in septic animals was only detected after 7 days in PL/PI treated animals. Activity of oxalacetic-glutamic transaminase was altered in all groups while glutamic-pyruvic transaminase not. Lactate dehydrogenase was augmented in all groups. Histopathological examination of liver and spleen revealed tissue damage caused by sepsis, but these alterations appeared later (7 days) in PL/PI-treated animals. Results reported in this study suggest that PL modulates immune inflammatory activity in septic animals by an indirect mechanism that remains to be investigated. Activity of LP leads to animal survival, even under infection. It is proposed that LP modulates NO synthesis, reducing NO levels in serum of infected animals and abolishing the failure of neutrophil migration. The pharmacological properties of LP previously described in classical models of inflammation, is now confirmed in a model of systemic inflammatory response elicited by microbial infection.
O látex da planta medicinal Calotropis procera (Apocynaceae) possui moléculas com diferentes atividades farmacológicas, dentre estas, pró- e antiinflamatória. Neste trabalho, o efeito imunomodulatório da fração de proteínas solúveis do látex (PL) foi investigado em modelos experimentais de sepse induzida por CLP e por inoculação de S. Typhimurium. Aspectos bioquímicos das proteínas do látex foram investigados e parâmetros bioquímicos, hematológicos e histopatológicos foram determinados em animais experimentais. PL exibiu um significativo efeito protetor nos animais. Uma dose de 30 mg/Kg levou à sobrevivência de até 100%, comparada com 100% de mortalidade no grupo de animais infectados e não tratados (grupo Salmonella). Este efeito foi somente observado quando PL foi administrado 24 horas antes da indução da sepse. PL não apresentou atividade antibacteriana in vitro sugerindo um mecanismo de ação indireto, possivelmente intervindo nos processos imunes. A população microbiana no fígado, baço e fluido peritoneal - local do foco infeccioso - estava similar entre todos os grupos, 4 e 24 horas após inoculação bacteriana. No sangue, entretanto, a quantidade de bactérias viáveis estava reduzida apenas nos animais tratados com proteínas do látex. O efeito protetor de PL foi revisto através de suas três sub-frações protéicas, denominadas de PI, PII e PIII, obtidas através de fracionamento por cromatografia de troca iônica em coluna de CM-Sepharose a pH 5,0. As três sub-frações protéicas apresentaram efeito protetor e de forma similar. Nem o tratamento térmico de PL (100 °C, 30 min.) ou inibição de suas proteases endógenas com iodoacetamida alterou o efeito protetor observado. O óxido nítrico (NO), um importante sinalizador molecular, estava aumentado no soro de animais do grupo Salmonella enquanto que animais protegidos com doses de PL e PI apresentaram níveis similares ao controle após 24 horas da infecção. Este resultado corroborou com a observação de que animais com sepse letal apresentaram falência na migração de neutrófilos para o foco infeccioso enquanto que, de forma evidente, animais tratados com PL ou PI favoreceram o influxo de células para a cavidade peritoneal. Além disto, nestes animais, o nível de NO estava aumentado no foco infeccioso, uma provável atividade dos neutrófilos presentes como parte de sua atividade microbicida. A atividade da adenosina desaminase (ADA) mensurada no foco infeccioso, após 4 e 24 horas de infecção, estava aumentada apenas nos animais tratados com PL ou PI, e inalterada em animais do grupo Salmonella. Plaquetopenia foi observada em animais com sepse, mas não em animais protegidos com PL ou PI. Neutrofilia e linfopenia ocorreram em animais do grupo Salmonella. Neutrofilia seria concordante com o elevado teor de NO, um inibidor da migração de neutrófilos, enquanto que a linfopenia é parte da imunossupressão estabelecida na sepse. Em animais tratados com PL ou PI foi observada neutrofilia sete dias após a infecção, sugerindo um estímulo na hematopoiese. Linfopenia, observada no início da sepse (24 h), só ocorreu aos sete dias nos animais tratados com PL ou PI. Atividade de transaminase glutâmico oxalacética estava aumentada em todos os grupos. Transaminase glutâmico pirúvica não teve atividade alterada. A atividade da lactato desidrogenase estava aumentada em todos os grupos. Análises histopatológicas do fígado e baço mostraram que os danos teciduais causados pela sepse foram retardados nos animais tratados com PL, como observado após sete dias. A análise integrada de todos os resultados sugere que as proteínas do látex modulam a resposta imunoinflamatória por mecanismo indireto, revertendo à falência da migração de neutrófilos causada pela sepse letal e, desta forma, induzem uma resposta imunológica ainda não esclarecida, que permite a sobrevivência dos animais, mesmo sob infecção. Os resultados sugerem que a modulação de NO estaria diretamente envolvida no efeito protetor final observado. As propriedades farmacológicas de PL previamente descritas em modelos clássicos de inflamação são agora confirmadas em um modelo de resposta inflamatória sistêmica resultante de um processo infeccioso previamente estabelecido.
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