ESTUDOS PRÉ-CLÍNICOS DE ATIVIDADE GIARDICIDA DE Chenopodium ambrosioides L. E A PADRONIZAÇÃO DOS EXTRATOS NA PESQUISA E DESENVOLVIMENTO DE FITOTERÁPICOS

Chenopodium ambrosioides L. (Chenopodiaceae), conhecida como erva-de-Santa-Maria ou mastruz, representa espécie exótica, de grande ocorrência no estado do Maranhão, muito utilizada na prática popular para diversos fins terapêuticos. Estudos farmacológicos têm comprovado o potencial da espécie para desenvolvimento de bioprodutos como alternativa terapêutica. Assim, o objetivo deste trabalho foi desenvolver metodologia analítica para padronização dos extratos de C. ambrosioides empregando ensaios químicos e biológicos de atividade anti-Giardia, visando o desenvolvimento de tecnologia farmacêutica na obtenção de novos fármacos no tratamento da giardíase. Foram obtidos extratos das folhas de C. ambrosioides por planejamento fatorial dos parâmetros: natureza do solvente (etanol 70% e água), operação de extração (maceração, percolação e extração por Soxhlet) e hidromódulo; submetidos a análise química e avaliação da atividade giardicida in vitro. Os resultados permitem concluir que a obtenção dos extratos hidroalcoólicos das folhas de C. ambrosioides pelo procedimento extrativo de percolação e maceração representam os extratos com melhor perfil químico e/ou atividade giardicida; evidenciando que a natureza do solvente, o procedimento extrativo e o hidromódulo são variáveis que  infuenciam na obtenção dos extratos da espécie em estudo.Descritores: Chenopodium ambrosioides; Padronização de Extratos; Atividade Giardicida; in vitro.Abstract: Chenopodium ambrosioides L., Chenopodiaceae, (herb-of-Santa-Maria or mastruz), represents high occurrence alien species, in the State of Maranhão, widely used in popular practice for many therapeutic purposes. Pharmacological studies have demonstrated the potential of species to develop therapeutic bioproducts such as new alternatives. Thus, the goal of this work was to developanalytical methodology for standardized extracts of Chenopodium. ambrosioides employing chemical and biological tests of giardicidal activity, aiming at the development of pharmaceutical technology in getting new drugs for Giardia treatment. Were obtained extracts of leaves of C. ambrosioides by factorial planning factors: nature of solvent (ethanol 70 and water), operation of extraction (maceration, percolation and Soxhlet extraction) and hydromodule (plant:solvent); subjected to chemical analysis and evaluation of giardicidal activity  in vitro. The results suggest that the obtaining of hydroalcoholic extracts from the leaves of C. ambrosioides extractive of percolation and maceration represents the extract with better chemical profle and/or giardicidal activity; evidencing that the nature of the solvent, extractive procedure and hydromodule are variables that infuence in obtaining extracts of the species under study.Descriptors: Chenopodium ambrosioides; Standardization of Extracts; Giardicidal Activity; in vitro

Effect of Arrabidaea Chica Verlot Hydroalcoholic Extract on Monosodium Iodoacetate-Induced Osteoarthritis of Rat Knees / Efeito do Extrato Hidroalcoólico de Arrabidaea chica Verlot na osteoartrite induzida por Monoiodoacetato de Sódio em joelhos de ratos

Introdution: The Arrabidaea chica Verlot (A. chica, ACV), with well-demonstrated anti-inflammatory properties,  appears as an option with therapeutic potential for the osteoarthritis; thus, validating its use is highly relevant. Method: 72 rats were allocated to 3 groups: control, osteoarthritis and phytotherapy {these last two were subjected to osteoarthritis induction, and treated orally with 0.9% normal saline (0.1 mL/100 g) and ACV hydroalcoholic extract (500 mg/kg), respectively, from days 7 to 28}. The 3 groups were subjected to weekly (days 7, 14, 21, 28) assessments including clinical tests (weight-bearing and von Frey), radiological and histopathological analyses.  Fractionation of the ACV´s hydroalcoholic extract was performed and it´s fractions were analysed. Results: The evaluation of the values of the osteoarthritis and phytotherapy groups showed significant difference, with p <0,05 : weight-bearing- on days 14 (29,64 x 35,52), 21 (32,62 x 42,53) and 28 (33,56 x 47,14), von Frey- on days 14 (31,12 x 37,80), 21 (30,24 x 41,48) and 28 (35,78 x 46,09),  x-ray- on days 21 (2,17 x 1,20) and 28 (2,33 x 1,40), and histopathological analysis- on day 28 (0,03 x 2,20). The fractionation of the extract obtained the FH (hexane), FC (chloroform), FAE (ethyl acetate) and FB (butanolic) fractions. The FAE had highest total polypnenolic contents and the FH had the highest concentration of total flavonoids. Conclusion: The ACV extract promoted a reduction in static incapacitance, allodynia, radiological score and degree of synovitis, and FAE and FH fractions are probably the fractions responsible for the anti-inflammatory and analgesic activities of the ACV extract

Prophylactic and Therapeutic Use of Strontium Ranelate Reduces the Progression of Experimental Osteoarthritis

Introduction: Strontium ranelate (SrRan) has the potential to interfere in the progression of osteoarthritis (OA), multifactorial disease associated with mechanical problems and articular inflammatory changes.Objectives: This study aimed to test the effects of prophylactic and therapeutic use of SrRan on clinical parameters of pain, the inflammatory process, and degradation of the articular cartilage.Methods: This was an experimental study, using a model of knee OA induced by intra-articular injection of monoiodoacetate. Thirty Wistar rats were divided into five groups and treated as indicated: control, without intervention; prophylactic, received SrRan at a daily oral dose of 250 mg/kg for 28 days before OA induction; SrRan treatments, administered 250 or 500 mg/kg/day for 28 days after the induction; and model control, received saline solution after the induction. Behavioral tests (joint incapacity, mechanical hyperalgesia, tactile sensitivity, and forced ambulation), histological evaluation of articular cartilage, and determination of inflammatory cytokines in the synovial fluid (interleukin [IL]-6, IL-10, tumor necrosis factor [TNF]-α, and interferon [INF]-γ) were performed.Results: Both prophylactic and therapeutic treatments improved the articular discomfort. A prophylactic dose of 500 mg/kg/day also improved mechanical hyperalgesia and the same dose was beneficial on tactile sensitivity. SrRan did not improve ambulation. Levels of IL-6, IL-10, TNF-α, and IFN-γ in SrRan-treated groups with OA were not significantly different compared with those in the normal control animals. The histopathological evaluation showed less articular damage in the SrRan-treated and control groups compared to the saline-treated group.Conclusion: The prophylactic and therapeutic administration of SrRan was associated with improved behavioral patterns of pain, especially joint discomfort. SrRan administration mitigated histological changes in the articular cartilage and reduced the inflammatory process, which beneficially reduced the progression of OA in the experimental model studied

AVALIAÇÃO DOS EFEITOS DAS ISOFLAVONAS DE SOJA EM RATAS COM HIPOESTROGENISMO INDUZIDO

O climatério é o período da vida feminina no qual o organismo sofre mudanças no metabolismo decorrentes da queda estrogênica, resultando em efeitos sobre os principais sistemas, influenciando na qualidade de vida e comportamento da mulher. Para tratamento, se utiliza a reposição hormonal de estrógeno. Contudo, essa opção tem trazido muitos efeitos colaterais, inclusive o desenvolvimento de neoplasias. Como alternativa a aliviar tais efeitos, têm sido utilizados fitoestrógenos, pois exercem ação agonista do hormônio em seus receptores sem causar os efeitos colaterais indesejados. Este trabalho tem como objetivo avaliar os efeitos das isoflavonas de soja sobre o útero, consumo alimentar, peso corporal e ciclo estral de ratas ooforectomizadas. Foram utilizadas ratas wistar submetidas à ooforectomia e por 90 dias foram tratadas diariamente de acordo com o grupo de estudo: ooforectomizadas tratadas com placebo,ooforectomizadas tratadas com isoflavonas de soja, ooforectomizadas tratadas com benzoato de estradiol e grupo falso-operado. As ratas tratadas com isoflavonas de soja tiveram o ciclo estral regular semelhante ao grupo falso-operado. Porém, apresentaram perda ponderal similar ao grupo do tratamento hormonal.Palavras-chave: Isoflavonas. Útero. Ratas. Hormonal.EVALUATION OF THE EFFECTS OF SOY ISOFLAVONES IN RATS WITH INDUCED HYPOESTROGENISMAbstract: The climacteric is the period of life in which the female body goes through changes in metabolism following a strogenic fall resulting in effects on the major systems influencing the quality of life and behavior of women. For treatment is used estrogen replacement therapy. Yet it has brought many side effects,including cancer development. As an alternative to alleviate such effects, phytoestrogens have been used because exert hormone agonist to its receptors without causing unwanted side effects. This study aims to evaluate the effects of soy isoflavones on the uterus, food intake, body weight and estrous cycle in ovariectomized rats. We used Wistar rats underwent oophorectomy and were treated for 90 days daily according to the study group: ovariectomized and treated with placebo, ovariectomized treated with soy isoflavones, ovariectomized and treated with estradiol benzoate and false-operated group. The rats treated with soy isoflavones had regular estrous cycle similar to the false-operated group. However, showed similar weight loss to the group of hormone treatment.Keywords: Isoflavones. Uterus. Rats. Hormone.EVALUACIÓN DE LOS EFECTOS DE LAS ISOFLAVONAS DE SOJA EN RATAS CON HIPOESTROGENISMO INDUCIDAResumen: El climaterio es el período de la vida en la que el cuerpo de la mujer pasa por cambios en el metabolismo como consecuencia de una caída estrogénica y afectando los sistemas principales que influyen en la calidad de vida y comportamiento de la mujer. Como tratamiento se emplea la terapia de reemplazo de estrógeno. Sin embargo, esto ha traído muchos efectos secundarios, incluyendo el desarrollo de cáncer. Como una alternativa para aliviar esas consecuencias han sido utilizados fitoestrógenos, los cuales ejercen una acción agonistas del hormonio en sus receptores, sin causar efectos secundarios no deseados. Este estudio tiene como objetivo evaluar los efectos de las isoflavonas de la soja en el útero, en la ingestión de alimentos, en el peso corporal y en el ciclo estral en ratas ovariectomizadas. Se utilizaron ratas Wistar que se sometieron a ovariectomización y fueron tratadas diariamente durante 90 días de acuerdo con el grupo de estudio: ovariectomizadas y tratadas con el placebo, ovariectomizadas tratadas con isoflavonas de soja, ovariectomizadas y tratadas con benzoato de estradiol y el grupo de falso-operado. Las ratas tratadas con isoflavonas de soja con presentaron un ciclo estral regular de manera semejante al grupo de falso-operado. Sin embargo, mostraron una pérdida de peso similar al grupo de tratamiento hormonal.Palabras clave: Isoflavonas. Útero. Ratas. Hormonal

Investigação dos efeitos toxicos e anti-hipertensivo de Arrabidaea chica Verlot (Bignoniaceae)

The species Arrabidaea chica (Bignoniaceae) is indicated to treatment of skin illnesses, renal stones, systemic arterial hypertension, as an antiinflammation and antianemia. In this paper it was avaliated the toxical effect of the extract of leaves of Arrabidaea chica Verlot (EAC), the chemical composition of EAC and the performance of EAC n the cardiovascular system of normotense and hypertense rats SHR. To the preparation of the EAC it was used dried pulverized leaves, macerated in ethanol 70%. After that the obstention of the EAC this was fractioned when it obtained several fractions among them the F63, this because of presenting in chromatrography compared and thinned. It was cleaner and in a major quantity. It was analysed in farmacological experiments, 1RMNH and CLAE. In the chemical composition of the EAC, it was detected the presence of flobafenical tanines, flavonos and steroidal componnds. In the the fraction F63 it was observed characteristic signals as to the alifatical componnds and aromatical. Among several possibilities they can beflavonoidal. To the avaliation of the acute toxicity (DL50) adult mice of both sexes received saline (0,1ml/10 g v. o) or EAC (0,5m 1,0 e 2,0 g/kg, ip), this didn t cause the death of animals as well it didn t get the behaviour alterations or macroscopic alterations of organs if compored to the control group. The treatment of rats with saline (0,1 mL/ 100g) and EAC (0,5,-5,0 g/kg, v.o), didn t cause deaths, nor behaviour or macroscopic alterations of the organs after antopsy. Finished the period of 14 days of treatment the blood was collected to its hematological and biochemical analysis, there wasn t difference between the control groups and treated in relation to the ration consume, corporal weight as well it wasn t detected alterations of behaviour or in the macroscopic analysis of the organs. The EAC (1,0 g/ kg) promoted meanful alteration in the VCM and in the doses of hemoglobine. The EAC of Arrabidaea chica Verlot (1 g/kg) promoted meanful alterations in the creatine, urea and cholesterol (HDL), in the dose of 0,5 g/Kg. It promoted, also, meanful alterations in the urea and in the HDL. The EAC (2,0 g/ kg) altered the glicosis, AST and total cholesterol, as to the dose of 3,5 g/Kg the EAC altered glicosis and total cholesterol. The EAC (5,0 g/kg) promoted alterations of glicosis, AST, urea and HDL the EAC (0,1; 0,2 e 0,5 g/kg, v.o.) administrated to hypertense rats SHR produced hypotension (38,2%; 49,9% and 34,4%) and bradicardia (13,4%; 15,35 % and 15,25%) as to the controls. In normotense animals there was reduction of the tension pressorical levls without alterations of the cardial frequence. In the menseterical superior arteries isolated of normotense rts or hypertense SHR submitted to chronical treatment with EAC for 60 days it was possible to detect reductions of the maximal effect and displacement of the curve of presence of noradrenalin when compared to the tissues of the respective control groups. In the analysis in vitro performed in rings of mesenterical superior artery isolated of rats with and without functional endothelial, in the presence and absence of the EAC (100-500 μg/mL) and F63 (10-30μg/mL) ocorred reduction of the maximal effect in the three used doses, in a way that the dependent dose and statistically meanful action in preparations of the mensenteric pre-contractd artery with moradrenalin, the EAC (10-500 μg/mL) and F63 (10-50μg/mL) promoted the dependent relaxing of dose both in the presence of the endotelial and in the absence of it. In the presence of L-NAME or indometacine, the EAC didn t promoted difference of the vasorelaxing. The EAC promoted a relaxing in precontracted preparations with KCI 80 mM bolh in the presence and in the absence vascular endotelial. The EAC (50-500 μg/mL) and F63 (10-30 μg/mL) reducted the maximal induced contractions per CaCl2. In precontracted preparations with S-(-)BayK 8644, the EAC (10-500 μg/mL) and F63 (10-300 μg/mL) promoted relaxing. The induced contractions per caffeine in a free ambience of calcium were inhibited by EAC, but the F63 didn t promoted alterations of the induced contractions by noradreline or caffeine. In a set these results suggest that the hypotension produced by the EAC of Arrabidaea chica Verlot can be realted to a diminution of the periferical total resistence in consequence of the participation of the bhlocade of the canaes CaV1.2 in cells of the vascular smooth muscle as well as the blocade of the rianodine channels.Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorA espécie vegetal Arrabidaea chica Verlot (Bignoniaceae) é indicada para o tratamento de enfermidades da pele, cálculos renais, hipertensão arterial sistêmica, como antiinflamatório e antianêmico. Neste trabalho avaliou-se extrato das folhas de Arrabidaea chica Verlot (EAC), a composição química do EAC, sua toxicidade e atividade no sistema cardiovascular de ratos normotensos e hipertensos SHR. Para a preparação do EAC usou-se folhas secas pulverizadas e maceradas em etanol 70%. Após a obtenção do EAC este foi fracionado obtendo-se várias frações, dentre as quais a F63, que por apresentar em cromatrografia comparativa delgada, mais limpa e em maior quantidade, foi analisada em experimentos farmacológicos, 1RMNH e CLAE. Na composição química do EAC, detectou-se a presença de taninos flobafênicos, flavononois, flavonononas e compostos esteroidais. Na fração F63, observou-se sinais característicos para compostos alifáticos e aromáticos que dentre várias possibilidade, podem ser flavonóides. Para a avaliação da toxicidade aguda (DL50) camundongos adultos, de ambos os sexos, receberam salina (0,1 ml/ 10g) ou EAC (0,5, 1,0 e 2,0 g/kg, ip), o que não produziu morte dos animais, bem como, não causou alterações comportamentais, ponderal ou alterações macroscópicas dos órgãos comparando-se ao grupo controle. O tratamento dos ratos com salina (0,1 mL/ 100g) e EAC não causou mortes, alterações comportamentais ou macroscópicas dos órgãos após autópsia. Ao final do período de tratamento de 14 dias, o sangue foi coletado para análise hematológica e bioquímica. Não houve diferença entre os grupos controle e tratado em relação ao consumo de ração, de água, peso corpóreo, bem como, não foram detectadas alterações de comportamento ou na análise macroscópica dos órgãos. O EAC (1,0 g/ kg) promoveu alteração significativa no VCM e na dosagem de Hemoglobina. O EAC de Arrabidaea chica Verlot (1 g/kg) promoveu alterações significativas na creatinina, uréia e HDL, na dose de 0,5 g/Kg, promoveu alteração significativa na uréia e no HDL. O EAC (2,0 g/ kg) alterou a glicose, AST e colesterol total, já na dose de 3,5 g/Kg o EAC alterou glicose e colesterol total. O EAC (5,0 g/kg) promoveu alteração de glicose, AST, uréia e HDL. O EAC (0,1; 0,2 e 0,5 g/kg, v.o.) administrado a animais SHR produziu hipotensão (38,2%; 49,9% e 34,4%) e bradicardia (13,4%; 15,35 % e 15,25%), com relação aos controles. Em animais normotensos houve redução dos níveis tensionais pressóricos sem alteração da freqüência cardíaca. Nas artérias mesentérica superior isoladas de ratos normotensos ou hipertensos SHR submetidos ao tratamento crônico com EAC por 60 dias foi possível detectar redução do efeito máximo e deslocamento da curva na presença de noradrenalina quando comparado aos tecidos dos respectivos grupos controles. Na análise in vitro realizadas em anéis de artéria mesentérica superior isolada de rato com e sem endotélio funcional, na presença e ausência do EAC (100- 500 μg/mL) e F63 (10 - 30μg/mL) ocorreu redução do efeito máximo nas três doses utilizadas, de maneira dose dependente e estatisticamente significante em preparações com endotélio intacto. Já preparações de artéria mesentérica pré-contraídas com noradrenalina, o EAC (10-500 μg/mL) e F63 (10-50μg/mL) promoveram relaxamento dependente de dose tanto na presença do endotélio quanto na ausência do mesmo. Na presença de L-NAME ou indometacina, o EAC não promoveu diferença do vasorelaxamento. O EAC promoveu um relaxamento em preparações pré-contraídas com KCl 80 mM, tanto na presença quanto na ausência do endotélio vascular. O EAC (50-500 μg/mL) e F63 (10-30 μg/mL) reduziram as contrações máximas induzidas por CaCl2. Em preparações pré-contraídas com S-(-)BayK 8644, o EAC (10-500 μg/mL) e F63 (10-300 μg/mL) promoveram relaxamento. As contrações induzidas por cafeína em meio livre de cálcio foram inibidas pelo EAC, porém a F63 não promoveu alterações das contrações induzidas por noradrenalina ou cafeína. Em conjunto, estes resultados sugerem que a hipotensão produzida pelo EAC de Arrabidaea chica Verlot pode estar relacionada a uma diminuição da resistência periférica total em conseqüência da participação do bloqueio dos canais CaV1.2 em células da musculatura lisa vascular, bem como, bloqueio dos canais de rianodina

Ethyl Acetate Fraction of Bixa orellana and Its Component Ellagic Acid Exert Antibacterial and Anti-Inflammatory Properties against Mycobacterium abscessus subsp. massiliense

Mycobacterium abscessus subsp. massiliense (Mabs) causes chronic infections, which has led to the need for new antimycobacterial agents. In this study, we investigated the antimycobacterial and anti-inflammatory activities of the ethyl acetate fraction of Bixa orellana leaves (BoEA) and ellagic acid (ElAc). In silico analysis predicted that ElAc had low toxicity, was not mutagenic or carcinogenic, and had antimicrobial and anti-inflammatory activities. Apparently, ElAc can interact with COX2 and Dihydrofolate reductase (DHFR) enzymes, which could explain both activities. In vitro analysis showed that BoEA and ElAc exerted antimicrobial activity against Mabs (minimum inhibitory concentration of 1.56, 1.56 mg/mL and bactericidal concentration of 6.25, 3.12 mg/mL, respectively. Clarithromycin showed MIC and MBC of 1 and 6 µg/mL). Treatment with BoEA or ElAc increased survival of Tenebrio molitor larvae after lethal infection with Mabs and reduced carrageenan-induced paw edema in mice, around 40% of edema volume after the fourth hour, similarly to diclofenac. In conclusion, BoEA and ElAc exert antimicrobial effects against Mabs and have anti-inflammatory effects, making them potential sources of antimycobacterial drugs. The biological activities of ElAc may be due to its high binding affinities predicted for COX2 and DHFR enzymes

“K-Powder” Exposure during Adolescence Elicits Psychiatric Disturbances Associated with Oxidative Stress in Female Rats

Ketamine, also called ‘K-powder’ by abusers, an analog of phencyclidine, primarily acts as an antagonist of N-methyl-D-aspartic acid (NMDA) receptors, therapeutically used as an anesthetic agent. Ketamine also stimulates the limbic system, inducing hallucinations and dissociative effects. At sub-anesthetic doses, ketamine also displays hallucinatory and dissociative properties, but not loss of consciousness. These behavioral consequences have elicited its recreational use worldwide, mainly at rave parties. Ketamine is generally a drug of choice among teenagers and young adults; however, the harmful consequences of its recreational use on adolescent central nervous systems are poorly explored. Thus, the aim of the present study was to characterize the behavioral and biochemical consequences induced by one binge-like cycle of ketamine during the early withdrawal period in adolescent female rats. Adolescent female Wistar rats (n = 20) received intraperitoneally administered ketamine (10 mg/kg/day) for 3 consecutive days. Twenty-four hours after the last administration of ketamine, animals were submitted to behavioral tests in an open field, elevated plus-maze, and forced swimming test. Then, animals were intranasally anesthetized with 2% isoflurane and euthanized to collect prefrontal cortex and hippocampus to assess lipid peroxidation, antioxidant capacity against peroxyl radicals, reactive oxygen species, reduced glutathione, and brain-derived neurotrophic factor (BDNF) levels. Our results found that 24 h after recreational ketamine use, emotional behavior disabilities, such as anxiety- and depression-like profiles, were detected. In addition, spontaneous ambulation was reduced. These negative behavioral phenotypes were associated with evidence of oxidative stress on the prefrontal cortex and hippocampus