Experimental acute anti-inflammatory activity of preparations with complexed cannabidiol in carriers

Abstract Cannabidiol (CBD) is a bioactive compound with promising anti-inflammatory results but has low aqueous solubility. Complexation of drugs with this characteristic in carriers is an alternative to improve their efficiency. This study aimed to prepare and characterize CBD complexes in different carriers, and to evaluate the anti-inflammatory effect of such preparations using an experimental model of edema induction in rat paws. The results were compared to a reference drug, ibuprofen (IBU). The carriers evaluated were beta cyclodextrin (bCD) and activated charcoal (AC). Quantification of the drugs in the complexes was determined, and different qualitative analyses were also performed. Oral treatments in single doses with CBD showed inhibitory effects similar to that of IBU, potentiating its bioactivity without significant adverse effects. CBD*bCD doses at 4.375, 8.75, 17.5, and 35 mg/kg significantly reduced the intensity of edema compared to equivalent doses of pure bioactive. In contrast, CBD*AC did not generate benefits. There was no significant inhibitory effect on myeloperoxidase activity, requiring more specific analyses to assess this parameter. The results suggest that the CBD*bCD complexation is perfectly feasible, increasing its anti-edematogenic efficacy in the experimental model used.

Immune response and Raman scattering assessment in rats skin after contact with Fusarium oxysporum metabolites.

The secondary metabolites produced by Fusarium can cause disease and death when consumed and produce biological responses even in the absence of the microorganism. The IL-6, TNF-α and TGF-ß1 cytokines immune reactivity was associated with histopathological and physico-chemical changes in skin of immune competent rats after administration of Fusarium oxysporum crude extract. Rats were intradermally injected with 50 µl of 0.5 mg/ml crude extract and were euthanized at 3, 6, 12 and 24 h after injection. The inflammatory response was quantified by enzyme myeloperoxidase activity and by immunohistochemical method to detect the IL-6, TNF-α and TGF-ß1. Physico-chemical analysis was performed using FT-Raman Spectroscopy. The inflammatory response was most intense at 6 and 12 h after crude extract administration and the most significant histopathological changes were observed in the dermis. Myeloperoxidase activity was intense from 3 to 24 h after injection. The immunostaining of pro-inflammatory cytokines IL-6 and TNF-α peaked at 6 h. Immunostaining for TGF-ß1 was highest at 12 and 24 h. FT-Raman spectral analysis showed both, the most intense Fusarium interaction with the skin at 6 h, as revealed by the changes in the stretching of -CH bands (3100-2800 cm-1) in the dermis, and skin recovery trending after 12 h after crude extract injection. The results showed that secondary metabolites stimulated histopathologic changes and inflammatory responses even in the absence of the fungus, increasing myeloperoxidase activity and pro-inflammatory cytokine expression besides promoting physico-chemical changes.

Evaluation of the effects of autohemotransfusion in arthritic rats induced by two doses of complete Freund's adjuvant

The aim the study was to evaluate the effects of autohemotransfusion in adjuvant-induced arthritis model by injections of high and low doses of Complete Freund ́s Adjuvant (CFA). Male Holtzman rats (200-230g) were distributed in six groups: control (C); control treated by autohemotransfusion (CT); CFA induced arthritis 0.5% w/v (AIA); CFA induced arthritis 0.5% w/v treated with autohemotransfusion (AIAT); CFA induced arthritis 0.1% w/v (AS) and CFA induced arthritis 0.1% w/v treated with autohemotransfusion (AST). The number of leukocytes, the weight of different organs and the paw volume were analyzed. The autohemotransfusion without erythrocytes promoted a reduction in the number of leukocytes in AIAT and AST when compared to AIA (p 0.05). The autohemotransfusion used in this work presented positive effects on AIA as they promoted a reduction in the number of leukocytes and an increase in thymus weight and body growth. However, other types of autohemotransfusion must be tested to determine the true efficacy of this alternative method of treatment.

Treatment with Trichilia catigua ethyl-acetate fraction improves healing and reduces oxidative stress in TNBS-induced colitis in rats.

Beverages containing Trichilia catigua are commonly employed in folk medicine. T. catigua bark extracts possess antioxidant, anti-inflammatory, and bactericidal properties. These properties suggest T. catigua bark extracts as a potential treatment for inflammatory bowel diseases (IBD). Using the 2,4,6-trinitrobenzenesulphonic acid (TNBS)-induced model of colitis in rats we evaluated the effect of an ethyl-acetate fraction (EAF) of T. catigua (200 mg/kg) administered by daily oral gavage or intrarectally at different time points after TNBS challenge. TNBS treatment evoked severe colonic inflammation after 24 h that persisted for 7 days, characterized by weight loss, high levels of myeloperoxidase activity, histological and macroscopic damage, and elevated index of oxidative stress in the blood. T. catigua EAF treatment prevented the oxidative stress within 24 h and enhanced tissue recovery observed at day 7, returning histological and macroscopic damage levels to that of the control group. TNBS treatment led to loss of myenteric neurons after 28 days. T. catigua EAF was unable to prevent the neuronal loss. Oral delivery of T. catigua EAF was more effective than intrarectal administration of the extract. In conclusion, T. catigua EAF treatment normalized oxidative stress parameters in blood and reduced the degree of acute inflammation in TNBS colitis.

Oxidative state and oxidative metabolism of the heart from rats with adjuvant-induced arthritis.

The aim of the present work was to investigate, in a more extensive way, the oxidative state and parameters related to energy metabolism of the heart tissue of rats using the model of adjuvant-induced arthritis. The latter is a model for the human arthritic disease. Measurements were done in the total tissue homogenate, isolated mitochondria and cytosolic fraction. The adjuvant-induced arthritis caused several modifications in the oxidative state of the heart which, in general, indicate an increased oxidative stress (+80% reactive oxygen species), protein damage (+53% protein carbonyls) and lipid damage (+63% peroxidation) in the whole tissue. The distribution of these changes over the various cell compartments was frequently unequal. For example, protein carbonyls were increased in the whole tissue and in the cytosol, but not in the mitochondria. No changes in GSH content of the whole tissue were found, but it was increased in the mitochondria (+33%) and decreased in the cytosol (-19%). The activity of succinate dehydrogenase was 77% stimulated by arthritis; the activities of glutamate dehydrogenase, isocitrate dehydrogenase and cytochrome c oxidase were diminished by 31, 25 and 35.3%, respectively. In spite of these alterations, no changes in the mitochondrial respiratory activity and in the efficiency of energy transduction were found. It can be concluded that the adjuvant-induced arthritis in rats causes oxidative damage to the heart with an unequal intracellular distribution. Compared to the liver and brain the modifications caused by arthritis in the heart are less pronounced on variables such as GSH levels and protein integrity. Possibly this occurs because the antioxidant system of the heart is less impaired by arthritis than that reported for the former tissues. Even so, the modifications caused by arthritis represent an imbalanced situation that probably contributes to the cardiac symptoms of the arthritis disease.

Constituintes químicos e avaliação das atividades antibacteriana e antiedematogênica de Aloysia gratissima (Gillies & Hook.) Tronc. e Aloysia virgata (Ruiz & Pav.) Pers., Verbenaceae / Chemical composition and evaluation of antibacterial and antiedematogenic Activities of Aloysia gratissima (Gillies & Hook.) Tronc. e Aloysia virgata (Ruiz & Pav.) Pers., Verbenaceae

O estudo químico de Aloysia gratissima (Gillies & Hook.) Tronc. e Aloysia virgata (Ruiz & Pav.) Pers., Verbenaceae, resultou no isolamento de quatorze substâncias. Suas estruturas foram determinadas com base nos dados de IV, RMN de ¹H e de 13C uni e bidimensionais e comparação com dados da literatura. A atividade antibacteriana do extrato bruto das folhas e caules e das frações de A. virgata, bem como as substâncias hoffmaniacetona e seu monoacetato, verbascosídeo e arenariosídeo, foram avaliadas através do método de bioautografia e a atividade antiedematogênica foi avaliada pelo modelo de edema de orelha induzido para os extratos brutos e frações metanólicas de A. gratissima e A. virgata..

Phytochemical study of Aloysia gratissima (Gillies & Hook.) Tronc. e Aloysia virgata (Ruiz & Pav.) Pers., Verbenaceae, afforded fourteen compounds. The structures were established using IR, 1D and 2D NMR and by comparison of its spectroscopic data to those of literature. The antibacterial activity of the crude extract from the leaves and branches and the fractions from the crude extract of A. virgata, besides the isolated compounds hoffmaniacetone, hoffmaniacetone monoacetate, verbascoside and arenarioside were evaluated by means of bioautography and the antiedematogenic activity was evaluated using induced ear oedema model to the crude extracts and methanol fractions for A. gratissima and A. virgata.

Sugar (sucrose) and fatty acid compounds with triglycerides on the treatment of wounds: experimental study in rats.

BACKGROUND: In the last 10 years, the use of essential fatty acids (EFA) compounds for the treatment of wounds has increased in Brazil, while there has been reducing indication for the use of sugar. OBJECTIVE: The aim of this study was to evaluate the response to saline, sugar and EFA on induced wounds in rats. METHODS: A wound of 400 mm(2) was produced on the back of each Wistar rat. The rats were divided into three groups, each being treated with either saline, sugar or EFA. All the animals received a closed dressing on the wounds, changed daily. Measures were taken in four moments, and the values of wound area reduction by healing, cellular inflammatory response, collagen ordering and types I and III collagen density were assessed. RESULTS: The wound healing was equal in all the three groups, but sugar promoted an inflammatory response modulation between the 7th and 14th days. On the 20th post-operative day, there were no differences between the three treated groups concerning types I and III collagen. CONCLUSIONS: The wounds healed in the three groups. The sugar group promoted effective cellular inflammatory response modulation. There were no differences between all the treated groups regarding types I and III collagen at the end of this study.

Açúcar (sacarose) e triglicerídeos de cadeia média com ácidos graxos essenciais no tratamento de feridas cutâneas: estudo experimental em ratos / Sugar (sucrose) and fatty acid compounds with triglycerides on the treatment of wounds: experimental study in rats

FUNDAMENTOS: Na última década, as indicações de uso tópico de compostos com ácidos graxos essenciais (AGE-TG) para o tratamento de feridas aumentaram no Brasil, e houve declínio das indicações do açúcar. OBJETIVOS: Estudar o efeito da aplicação de solução fisiológica de cloreto de sódio a 0,9 por cento, de açúcar e de AGE-TG sobre feridas experimentalmente induzidas em ratos. MÉTODOS: Foi induzida uma ferida de 400 mm2 no dorso de cada rato Wistar, constituindo três grupos tratados separadamente com solução fisiológica de cloreto de sódio a 0,9 por cento, açúcar e AGE-TG. Todos os animais receberam curativo oclusivo sobre a lesão, trocado a cada 24 horas. As aferições realizadas em quatro momentos consistiram na determinação do percentual de redução das áreas das feridas, da reação inflamatória celular, da ordenação do colágeno e da densidade de colágeno dos tipos I e III nas cicatrizes. RESULTADOS: A cicatrização ocorreu igualmente nos grupos estudados, mas o açúcar modulou positivamente a reação inflamatória entre o 7º e 14º dias. No 20º dia, não houve diferenças na quantidade de colágeno dos tipos I e III entre os grupos tratados. CONCLUSÕES: As feridas cicatrizaram nos três grupos. O grupo açúcar apresentou uma modulação positiva da resposta inflamatória celular. Não houve diferenças na quantidade de colágeno dos tipos I e III ao final do experimento nos grupos tratados.

BACKGROUND: In the last 10 years, the use of essential fatty acids (EFA) compounds for the treatment of wounds has increased in Brazil, while there has been reducing indication for the use of sugar. OBJECTIVE: The aim of this study was to evaluate the response to saline, sugar and EFA on induced wounds in rats. METHODS: A wound of 400 mm2 was produced on the back of each Wistar rat. The rats were divided into three groups, each being treated with either saline, sugar or EFA. All the animals received a closed dressing on the wounds, changed daily. Measures were taken in four moments, and the values of wound area reduction by healing, cellular inflammatory response, collagen ordering and types I and III collagen density were assessed. RESULTS: The wound healing was equal in all the three groups, but sugar promoted an inflammatory response modulation between the 7th and 14th days. On the 20th post-operative day, there were no differences between the three treated groups concerning types I and III collagen. CONCLUSIONS: The wounds healed in the three groups. The sugar group promoted effective cellular inflammatory response modulation. There were no differences between all the treated groups regarding types I and III collagen at the end of this study.

Action of Canova® medicine onto peritoneal resident macrophages infected with Trypanosoma cruzi / Ação do medicamento Canova® em macrófagos peritoniais residentes infectados por Trypanosoma cruzi

Approximately 20 million of people are chronically infected with Trypanosoma cruzi in Latin America. The present work investigated the action of the homeopathic medicine Canova® in in vitro experimental infections with T.cruzi, type Y, using Swiss mice peritoneal resident macrophages. Our results demonstrated that Canova® induced a decrease in the production of H2O2 and TNF-alpha at 20% and 40% concentrations when compared to the control RPMI. However, when compared with this medicine excipient, there was a significant decrease of these nediators at 40% concentration only. The production of NO and the phagocytic activity were not affected. TNF-alpha inhibits the T.cruzi replication in peritoneal macrophages in vitro, becoming an important agent of infection control by this parasite. Within this context, Canova®, unlike what has been reported to other infections, would function as a stimulator of the infection, since it inhibited the production of TNF-alpha by peritoneal resident macrophages in vitro. Further studies should be carried out with elicited macrophages, in order to confirm the Canova® inhibitory activity on the production of TNF-alpha and other mediators in macrophages infected by T.cruzi.

Aproximadamente 20 milhões de pessoas são cronicamente infectadas pelo Trypanosoma cruzi na América Latina. O presente trabalho investigou a ação do medicamento homeopático Canova® em infecções experimentais in vitro com Trypanosoma cruzi, cepa Y, usando macrófagos residentes peritoniais de ratos Swiss. Os resultados mostraram que Canova® induz uma diminuição significativa da produção de H2O2 e TNF-alfa em concentrações de 20% e 40%, quando comparado com o controle RPMI. Quando comparado com o excipiente do medicamento, observou-se uma diminuição na concentração destes mediadores apenas na concentração de 40%. A produção de NO e a atividade fagocítica não foram afetadas. TNF-alfa inibe a replicação do protozoário em macrófagos peritoniais in vitro, mostrando-se um importante agente para o controle da infecção pelo parasita. Portanto, o medicamento Canova® poderia estimular o processo de infecção, pois promoveu inibição da produção de TNF-alfa por macrófagos peritoniais residentes in vitro. Estudos adicionais devem ser realizados com macrófagos elicitados, a fim de confirmar a atividade inibitória da Canova® sobre a produção de TNF-alfa e outros mediadores em macrófagos infectados por T. cruzi.

Prevalência de risco cardiovascular em pacientes que fazem uso de terapia anti-retroviral / Prevalence of cardiovascular risk in patients who make use of antiretroviral therapy

As manifestações cardiovasculares nos pacientes HIV são as mais diversas, conseqüentes da própria infecção pelo HIV, auto-imunidade, reação imunológica diante de outras infecções virais, inflamação crônica, neoplasias, imunossupressão prolongada, desnutrição e cardiotoxicidade dos medicamentos. Este trabalho teve como objetivo avaliar a prevalência de risco cardiovascular em uma população HIV positiva de Maringá, Paraná, Brasil. Foram selecionados 60 pacientes por meio de amostragem sistemática. A avaliação clínica baseou-se em exame clínico geral com aferição de pressão arterial, pulso, freqüência cardíaca, peso corporal, estatura, medida de cintura e quadril. O perfil laboratorial foi obtido por meio das dosagens de glicose sérica em jejum e lipidograma (Colesterol total, LDL colesterol, HDL colesterol, triglicérides). Utilizou-se a tabela de Escores de Framingham para cálculo do Risco de Doença ArterialCoronariana (DAC). Com relação aos fatores de risco aparece maior prevalência de história familiar em 80%, sedentarismo em 45% e tabagismo em 35%, dislipidemia em 36,6%. Nenhum paciente apresentou risco eminente de DAC, 34% risco > 20% em 10 anos, 25% risco < 20% e >5% em 10 anos, e 71,6% risco <5% em 10 anos. Apesar dos dados apontarem o risco relativo de doençascardiovasculares, o risco absoluto permanece baixo para a maioria dos pacientes, tornando-se significativo naqueles com múltiplos outrosfatores de risco cardiovascular.

The cardiovascular manifestations in HIV patients are the widely diversed, consequent of the HIV infection itself, autoimmunity, imunological reaction ahead other viral infections, chronic inflammation, neoplasms, prolonged immunosupression, malnutrition and the cardiotoxicity of medicines. This work had as objective to evaluate the prevalence of cardiovascular risk of a HIV positive population from Maringá, Paraná, Southern Brazil. There were selected 60 patients by systematic sample. The clinical evaluation were based in general clinical exam with arterial pressure gauging, pulse, cardiac frequency, body weight, stature, measure of waist and hip. The laboratorial profiles were obtained by dosages of seric glucose fasting, lipidogram (total cholesterol, LDL cholesterol, HDL cholesterol, triglycerides). It was used the table of Framingham Scores to calculate the Coronary Artery Disease (CAD) Risk. In relation to the risk factors, which appear with major prevalence are family history in 80%, sedentarism 45%, and tabagism 35%, dislipdemy 36,6%. No patients presented imminent risk of CAD, 34% presented risk > 20% in 10 years, 25% risk < 20% and >5% in 10 years, 71.6% presented risk <5% in 10 years. Although the data points a relative risk of cardiovascular diseases, the absolute risk remains low to the majority of the patients, becoming significant in those with multiple factors of cardiovascular risk.

Glycogen levels and energy status of the liver of fasting rats with diabetes types 1 and 2

Glycogen levels and the energy status of livers from fasting rats with diabetes types 1 and 2 were measured. After a 24 h fast, the hepatic glycogen levels of rats with diabetes1 and diabetes2 were, 18.7 and 2.6 times higher, respectively, than those of livers from the normal rats. In diabetes1 rats, the glycogen levels decreased when the fasting period was extended to 48 and 72 h. The opposite occurred with the control and diabetes2 rats. Consistently, glucose release by the perfused livers from diabetes1 rats was considerably higher during at least 60 minutes after initiating perfusion. The hepatic ATP content of diabetes1 rats was similar to that of the control rats; in diabetes2 rats, the hepatic ATP content was increased. It could be concluded that regulation of glycogen deposition and degradation in rats with diabetes1 differed markedly from that of rats with diabetes2 which, in turn, behaved similarly to normal healthy rats.

Teores de glicogênio e os estados energéticos de fígados de ratos com diabete dos tipos 1 e 2 foram medidos. Após um jejum de 24 horas os teores de glicogênio de ratos com diabete1 e diabete2 foram, respectivamente 18,7 e 2,6 vezes superiores àqueles de fígados de animais controle. Em ratos com diabete1 o conteúdo de glicogênio diminuiu quando o período de jejum foi prolongado para 48 e 72 horas. O oposto ocorreu em ratos controle e ratos com diabete2. Consistentemente, a liberação de glicose por fígados em perfusão isolada obtidos de ratos com diabete1 foi consideravelmente maior durante ao menos 60 minutos após o início da perfusão. O conteúdo hepático de ATP de ratos com diabete1 foi similar àquele de ratos controle; em ratos com diabete2 o conteúdo hepático de ATP foi maior. Pode-se concluir que a regulação da deposição e degradação do glicogênio em ratos com diabete1 difere marcadamente daquela de ratos com diabete2, os quais, por seu turno, comportam-se similarmente a ratos normais e saudáveis.

Consumo de antiinflamatórios esteróides em farmácia comunitária / Consumption of steroidal anti-inflammatory drugs in a community pharmacy

O objetivo do presente estudo foi avaliar o consumo de antiinflamatórios esteróides em uma farmácia comunitária da cidade de Umuarama-Pr. Os dados foram obtidos através de entrevistas utilizando um questionário específico, a fim de avaliar o consumo de medicamentos com e sem prescrição, relacionado com os sintomas mencionados pelos usuários e a via de administração dos antiinflamatórios consumidos. Os resultados mostraram um alto consumo de antiinflamatórios esteróides, principalmente sem prescrição, indicando a necessidade de uma discussão mais ampla sobre a orientação ao paciente que procura a farmácia comunitária nestas circunstâncias, assim como o papel do farmacêutico nesse serviço de saúde

Modelo experimental de hérnia ventral em ratos / Experimental model of ventral hernia in rats

O objetivo deste experimento foi estudar o comportamento de defeitos provocados na parede abdominal, com a finalidade de estabelecer um modelo de hérnia ventral em ratos. Cem ratos com peso entre 230 e 260 g e idade de três meses foram distribuídos em dois grupos. No grupo I, os animais foram submetidos a uma incisão padronizada na linha alba. No grupo II, os animais foram submetidos à exérese padronizada músculo-aponeurótica da região anterior do abdome. Em ambos os grupos, os animais foram redistribuídos em três subgrupos (A, B e C), conforme o tempo de observação de 15, 30 e 45 dias, respectivamente. Nas hérnias desenvolvidas, foram analisados dimensões lineares e volume, presença de aderências e aspectos histológicos de fragmentos do saco herniário. Para análise de resultados, foram empregados testes estatísticos. Os animais dos subgrupos I A, I B e I C, respectivamente, 93,3 por cento, 45 por cento e 53,3 por cento, e dos subgrupos II A, II B e II C, respectivamente, 66,7 por cento, 35 por cento e 47,7 por cento, desenvolveram hérnias. Em ambos os grupos, as medidas do eixo maior dos ânulos herniários ultrapassaram 3 centímetros e os volumes, em média, superiores a 3 mL, classificando-as como hérnias de grandes dimensões. Aderências foram encontradas praticamente em todos os animais. A mortalidade pós-operatória foi em média de 8 por cento, no grupo I, e de 18 por cento, no grupo II, sendo a evisceração a causa dos óbitos. O estudo histológico mostrou maturação do saco herniário aos 30 dias de observação em ambos os grupos. De acordo com os resultados obtidos, o modelo incisão (grupo I) foi o mais adequado para simular hérnia ventral no rato, apesar de não ter havido desenvolvimento de hérnia em todos os animais

Infecções unrinárias / Urinary infections

As infecções do trato urinário podem ser definidas como a presença ou a colonizaçäo de bactérias na urina levando à infecçäo das estruturas do aparelho urinário. Os estudos sobre o tratamento destas infecções despertam grande interesse, pois estäo entre as enfermidades mais freqüentes na prática médica. Seu tratamento engloba näo só a eliminaçäo das bactérias presentes na urina e conseqüente cura bacteriológica, como também a elucidaçäo e prevençäo de fatores predisponentes e proteçäo da funçäo renal. Embora tenha ocorrido nos últimos anos grande avanço no desenvolvimento de novos antimicrobianos, com o surgimento de agentes mais potentes, eficazes e seguros, a melhor opçäo para infecçäo ainda é näo tê-la. Desta forma, as medidas preventivas exercem um papel fundamental no controle desta patologia