Evaluación del desempeño de una prueba rápida para el tamizaje de la infección por Trypanosoma cruzi en pacientes con el virus de la inmunodeficiencia humana / Performance evaluation of a rapid test for the screening of Trypanosoma cruzi infection in patients with human immunodeficiency virus

Resumen Un diagnóstico rápido y seguro de la infección por Trypanosoma cruzi permite la administración inmediata de un tratamiento etiológico específico, en los casos clínicamente manifiestos. En el presente trabajo, en 100 sueros de pacientes con diagnóstico presuntivo de infección por T. cruzi, se evaluó el desempeño de la prueba rápida Chagas Ab Rapid SD Bioline (PDR) para el diagnóstico serológico, se la comparó con el estándar diagnóstico (par serológico) y se la valoró para su empleo en la rutina del laboratorio. La PDR reveló un índice de concordancia muy bueno respecto del estándar diagnóstico (Kappa=0,989 [IC95% 0,965-1,000]) y los parámetros de sensibilidad, especificidad, valores predictivos positivos y negativos revelaron un buen desempeño. El índice de Youden informó un buen rendimiento de la prueba (J=0,98 [IC95% 0,93-1,02]); y en el mismo sentido, tanto la razón de verosimilitud positiva (RV (+)= infinito), como la negativa (RV (-)= 0,02 [IC95% 0,00-0,16]), mostraron aumentada la probabilidad que la enfermedad blanco esté presente o ausente, cuando la técnica así lo determinaba. El empleo de una PDR, por su simpleza y buen desempeño (con resultados disponibles el mismo día) produciría una optimización de recursos, podría realizarse en lugares donde el acceso al diagnóstico es limitado y donde su incorporación no requeriría de una infraestructura importante. Por otro lado, en nuestro caso, podría ser utilizada como segunda prueba, para incrementar la especificidad del diagnóstico serológico de la infección por T. cruzi.

Abstract A quick and safe diagnosis of Trypanosoma cruzi infection allows the immediate etiological treatment in clinically manifested cases. In the present work, the performance of the rapid Chagas Ab Rapid SD Bioline (PDR) test for serological diagnosis was evaluated in 100 sera from patients with a presumptive diagnosis of T. cruzi infection, it was compared with the diagnostic standard (serological pair) and this technique was valued to be used within the laboratory routine. The PDR revealed a very good concordance index with respect to the diagnostic standard (Kappa=0.989 [95% CI 0.965-1.000]) and the parameters of sensitivity, specificity, positive and negative predictive values revealed good performance. The Youden index reported good performance of the diagnostic test (J = 0.98 [95% CI 0.93-1.02]); and in the same sense, both the positive likelihood ratio (RV (+) = infinity), and the negative (RV (-) = 0.02 [95% CI 0.00-0.16]) showed an increased probability whether the target disease was present or absent, when the technique so determined. The use of a PDR, due to its simplicity and good performance (with results available on the same day) would produce an optimization of resources, could be carried out in places where access to the diagnosis is limited and where its incorporation would not require an important infrastructure. On the other hand, in our case, it could be used as a second test, to increase the specificity of the serological diagnosis of T. cruzi infection.

Resumo Um diagnóstico rápido e seguro da infecção pelo Trypanosoma cruzi permite a administração imediata de um tratamento etiológico específico nos casos clinicamente manifestos. No presente trabalho, o desempenho do teste rápido Chagas Ab Rapid SD Bioline (PDR) para o diagnóstico sorológico foi avaliado em 100 soros de pacientes com diagnóstico provável de infecção por T. cruzi, foi comparado com o padrão diagnóstico (par sorológico) e essa técnica foi avaliada para ser utilizada na rotina do laboratório. O PDR revelou um índice de concordância muito bom em relação ao padrão diagnóstico (Kappa=0,989 [IC 95% 0,965-1.000]) e os parâmetros de sensibilidade, especificidade, valores preditivos positivos e negativos revelaram bom desempenho. O índice de Youden informou um bom desempenho do teste diagnóstico (J = 0,98 [IC 95% 0,93-1,02]); e, no mesmo sentido, tanto a razão de verossimilhança positiva (RV (+) = infinito), quanto a negativa (RV (-) = 0,02 [IC 95% 0,00-0,16]), mostraram uma probabilidade aumentada de a doença-alvo estar presente ou ausente, quando a técnica assim o determinar. A utilização de um PDR, pela sua simplicidade e por um bom desempenho (com resultados disponíveis no mesmo dia) produziria uma otimização de recursos, podendo ser realizado em locais onde o acesso ao diagnóstico é limitado, e onde a sua incorporação não exigiria um infraestrutura importante. Por outro lado, no nosso caso, poderia ser utilizado como segundo teste, para aumentar a especificidade do diagnóstico sorológico da infecção pelo T. cruzi.

Citotoxicidade e efeito indutor de mineralização de flavonoides sobre células osteoblásticas / Cytotoxicity and inducing effect of mineralization of flavonoids on osteoblastic cells

A perda óssea dentária e a formação de lesões periapicais surgem como uma consequênc ia do desequilíbrio da homeostase óssea. Os osteoblastos, juntamente com os osteoclastos e osteócitos, atuam na formação e na reabsorção óssea. Vários marcadores de formação óssea são produzidos por osteoblastos ativos e refletem diferentes aspectos da dif erenciação osteoblástica e da remodelação óssea. Com isso, muitos autores têm explorado o uso de fitoterápicos, visando obter novos compostos que apresentem propriedades terapêuticas, como os flavonoides, e que estimulem a neoformação óssea e o reparo da r egião periapical. O objetivo deste estudo foi avaliar in vitro a citotoxicidade e efeito indutor de mineralização de flavonoides sobre células osteoblásticas humanas. Para isso, células osteoblásticas da linhagem Saos expostas aos seguintes flavono2 foram ides: quercetina, miricetina e seus derivados taxifolina, isoquercitrina, rutina, ampelopsina e EGCG, além de pinocembrina, crisina e canferol, de forma isolada e combinada. Foi avaliado o efeito citotóxico, a atividade de fosfatase alcalina e indução de n mé todo de Shapiroódulos de mineralização. Os resultados foram analisados p elo Wilk, e as variáveis foram submetidas à análise de ANOVA seguida pelo teste de Tukey para comparar entre os grupos e/ou concentrações ou teste de Dunnett para comparar entre cada grupo e o controle, com nível de significância de 5%. A viabilidade da cultura de osteoblastos não teve uma redução estatisticamente significativa na presença da maioria dos compostos, exceto crisina a 100µM. Taxifolina, isoquercitrina, rutina, ampelopsina e EGCG foram os compostos que estimularam significativamente a atividade da fosfatase alcalina, juntamente com as combinações taxifolina+isoquercitrina, taxifolina+ampelopsina e taxifolina+rutina a 25/25 µM. Quanto a formação de nódulos de mine ralização, ampelopsina, isoquercitrina, rutina, pinocembrina e miricetina isolados e taxifolina+isoquercitrina, taxifolina+ampelopsina e taxifolina+rutina combinados obtiveram os melhores resultados, variando de acordo com as concentrações. Concluise que a taxifolina, isoquercitrina, rutina e ampelopsina e combinações de taxifolina com esses flavonoides são citocompatíveis e apresentam efeito indutor de mineralização em osteoblastos Saos-2(AU)

Dental bone loss and the formation of periapical lesions arise as a consequence of imbalance of bone homeostasis. Osteoblasts, together with osteoclasts and osteocytes, act in bone formation and resorption. Several markers of bone formation are produced by active osteoblasts and reflect different aspects of osteoblastic differentiation and bone remodeling. Thus, many authors have explored the use of phytotherapics in order to obtain new compounds with therapeutic properties, such as flavonoids, and also stimulate bone neoformation and periapical region repair. The objective of this study was to evaluate in vitro the cytotoxicity and inducing effect of flavonoid mineralization on human osteoblastic cells. For this, osteoblastic cells of the Saos-2 lineage were exposed to the following flavonoids: quercetin, myricetin and its derivatives taxifoline, isoquercitrin, rutin, ampelopsin and EGCG, in addition to pinocembrin, chrysin and kaempferol, in an isolated and combined manner. The cytotoxic effect, the activity of alkaline phosphatase and the induction of mineralization nodules were evaluated. The results were analyzed using the Shapiro-Wilk method, and the variables were submitted to ANOVA analysis followed by the Tukey test to compare between groups and/or concentrations or Dunnett's test to compare between each group and the control, with a level of 5% significance. The viability of the osteoblast culture did not have a statistically significant reduction in the presence of most compounds, except 100 µM chrysin. Taxifoline, isoquercitrin, rutin, ampelopsin and EGCG were the compounds that significantly stimulated the activity of alkaline phosphatase, together with the combinations taxifoline+isoquercitrin, taxifoline+ampelopsin and taxifoline+rutin at 25/25 µM. As for the formation of mineralization nodules, ampelopsin, isoquercitrin, rutin, pinocembrin and myricetin alone and taxifoline+isoquercitrin, taxifoline+ampelopsin and taxifoline+rutin combined obtained the best results, varying according to the concentrations. It is concluded that taxifoline, isoquercitrin, rutin and ampelopsin and combinations of taxifolin with these flavonoids are cytocompatible and have a mineralization-inducing effect on Saos-2 osteoblasts(AU)

Research on Q-markers of Eupatorium lindleyanum based on analytic hierarchy process-entropy weight method and network pharmacology / 中国中药杂志

The potential quality markers( Q-markers) of Eupatorium lindleyanum were studied with analytic hierarchy process(AHP)-entropy weight method(EWM) and network pharmacological method. Based on the concept of Q-markers of traditional Chinese medicine, AHP-EWM was employed to quantitatively identify the Q-markers of E. lindleyanum. AHP method was applied to the weight analysis of the validity, testability, and specificity of the first-level indexes, and EWM method was used to analyze the secondlevel indexes supported by literature and experimental data. At the same time, based on the theory and method of network pharmacology, the component-target-disease-efficacy network of E. lindleyanum was built, and the components most closely related to the efficacy of resolving phlegm and relieving cough and asthma were screened out. Through the integrated analysis of the results obtained with AHP-EWM and network pharmacological method, 13 compounds including rutin, quercetin, nepetin, cirsiliol, luteolin, hyperoside,isoquercitrin, kaempferol, caffeic acid, eupalinolide K, eupalinolide A, eupalinolide B, and eupalinolide C were comprehensively identified as the potential Q-markers of E. lindleyanum. The results provide a basis for the quality control of E. lindleyanum.

The effect of mild hypothermia plus rutin on the treatment of spinal cord injury and inflammatory factors by repressing TGF-β/smad pathway

ABSTRACT Purpose To probe the mechanism of mild hypothermia combined with rutin in the treatment of spinal cord injury (SCI). Methods Thirty rats were randomized into the following groups: control, sham, model, mild hypothermia (MH), and mild hypothermia plus rutin (MH+Rutin). We used modified Allen's method to injure the spinal cord (T10) in rats, and then treated it with MH or/and rutin immediately. BBB scores were performed on all rats. We used HE staining for observing the injured spinal cord tissue; ELISA for assaying TNF-α, IL-1β, IL-8, Myeloperoxidase (MPO), and Malondialdehyde (MDA) contents; Dihydroethidium (DHE) for measuring the reactive oxygen species (ROS) content; flow cytometry for detecting apoptosis; and both RT-qPCR and Western blot for determining the expression levels of TGF-β/Smad pathway related proteins (TGF-β, Smad2, and Smad3). Results In comparison with model group, the BBB score of MH increased to a certain extent and MH+Rutin group increased more than MH group (p < 0.05). After treatment with MH and MH+Rutin, the inflammatory infiltration diminished. MH and MH+Rutin tellingly dwindled TNF-β, MDA and ROS contents (p < 0.01), and minified spinal cord cell apoptosis. MH and MH+Rutin could patently diminished TGF-β1, Smad2, and Smad3 expression (p < 0.01). Conclusions MH+Rutin can suppress the activation of TGF-β/Smad pathway, hence repressing the cellular inflammatory response after SCI.

Effect of drying processing methods on different specifications of Sophorae Flos based on comprehensive statistical analysis / 中国中药杂志

To investigate the effects of six common drying methods on the quality of different specifications of Sophorae Flos, in order to select their suitable drying methods. According to appearance and morphology, Sophorae Flos was divided into the following three specifications: flower bud type(HL), half-open type(BK) and blooming type(SK). All specifications of samples were treated with shade-drying method(25 ℃, natural temperature), sun-drying method, hot-air-drying method(60, 105 ℃), and drying method(60 ℃) after steaming. The contents of total flavonoids, rutin, narcissus, quercetin, isorhamnetin, and Fe~(3+) reducing ability, DPPH free radical scavenging ability, ABTS free radical scavenging ability and fluorescence recovery after photobleaching(FRAP) were detected by UV, HPLC and colorimetry, respectively. Principal component analysis(PCA), cluster analysis(CA) and correlation analysis were used to comprehensively evaluate the quality of samples. According to the results, there were significant differences in the effect of drying methods on different specifications of samples. The drying method(60 ℃) after steaming was suitable for HL and BK, while the hot-air-drying method(60 ℃) was suitable for SK. When the fresh medicinal materials could not be treated in time, they should be spread out in a cool and ventilated place. Under high and low temperature conditions, the quality of three specifications of Sophorae Flos would be reduced. The hot-air-drying method(105 ℃) and shade-drying method(25 ℃) were not suitable for the treatment of fresh flowers and flower buds of Sophora japonicus. There were obviously differences of chemical compositions and antioxidant activities among the three specifications of samples. Therefore, the specifications of medicinal materials should be controlled to ensure the uniform quality. The study provided the abundant data reference for the selection of appropriate drying methods for the three specifications of Sophorae Flos, and useful exploration for the classification and processing of medicinal materials of flowers.

Research on chemical constituents from Artemisia annua Ⅰ / 中国中药杂志

Chemical constituents were isolated and purified from the water extract of Artemisia annua by column chromatography of HP-20 macroporous resin, silica gel, ODS, Sephadex LH-20, HW-40, and semi-preparative RP-HPLC. Their structures were elucidated by physicochemical properties and spectral analyses. As a result, Fifteen compounds were isolated and identified as vitexnegheteroin M(1), sibricose A5(2), securoside A(3), citrusin D(4), annphenone(5), E-melilotoside(6), esculetin(7), scopoletin-7-O-β-D-glucoside(8), eleutheroside B_1(9), chrysosplenol D(10), patuletin-3-O-β-D-glucopyranoside(11), quercetin-7-O-β-D-glucoside(12), rutin(13), apigenin 6,8-di-C-β-D-glucopyranoside(14), isoschaftoside(15), among them, compounds 1-4 were identified from Artemisia for the first time. Additionally, the isolates were evaluated for their inhibitory effects on the production of PGE_2 in LPS-simulated RAW264.7 macrophages. The results showed that compounds 1, 2, 8, and 10-15 could reduce PGE_2 levels, to a certain extent.

Research progress on rutin derivatives / 中国中药杂志

Compounds derived from natural products present satisfactory efficacy in disease prevention and treatment. The use of chemical substances in plants to promote healthhas increasingly attracted people's attention. Rutin, a typical flavonoid, is mainly found in various vegetables, fruits and Chinese herbal medicines. As a natural antioxidant, it features many pharmacological activities, such as anti-inflammation, anti-virus, anti-tumor, and prevention and treatment of cardiovascular and cerebrovascular diseases. However, the low bioavailability and poor water solubility limit its clinical application. In view of this, its structure is optimized and modified to afford rutin derivatives with good solubility, high bioavailability, stable metabolism and small toxic side effects. So far, a large number of rutin ethers, esters, and complexes have been synthesized and undergone activity testing. This paper reviews the structural modification of rutin in recent years, and the obtained derivatives have excellent properties and significant biological activity.

Enzymatic synthesis of acylated quercetin 3-O-glycosides: a review / 生物工程学报

Quercetin 3-O-glycosides (Q3Gs) are important members of quercetin glycosides with excellent pharmacological activities such as anti-oxidation, anti-inflammation, anti-cancer and anti-virus. Two representatives of Q3Gs, rutin and troxerutin, have been developed into clinical drugs, demonstrating Q3Gs have become one of the important sources of innovative drugs. However, the applications of Q3Gs in food and pharmaceutical industries are hampered by its poor bioavailability. Of the known means, selective acylation modification of Q3Gs through enzymatic catalysis to obtain Q3G esters is one of the effective ways to improve its bioavailability. Herein, the enzyme-mediated acylation of Q3Gs were reviewed in details, focusing on the four tool enzymes (acyltransferases, lipases, proteases and esterases) and the whole-cell mediated biotransformation, as well as the effect of acylations on the biological activities of Q3Gs. Furthermore, the highly efficient synthesis and diversification of acylated site for Q3G esters were also discussed. Taken together, this review provides a new perspective for further structural modifications of Q3Gs towards drug development.

Suspensão oral de rutina interfere na hiperplasia hepática em ratos / Oral rutin suspension intervene in hepatic hyperplasia in rats

ABSTRACT BACKGROUND: Rutin is a flavonol glycoside that can be found in a wide variety of vegetables and has activity, anti-cancer, anti-inflammatory and anti-diabetic properties. OBJECTIVE: This study investigated the effect of rutin oral administration on Wistar rats submitted to hepatic hyperplasia after partial hepatectomy (PH). METHODS: To achieve this, we considered the analysis of hepatic hyperplastic and plasma biochemical activity of Wistar rats, subjected to treatment with rutin 40 mg/kg/day for 10 days in group 1 (G1) or saline in group 2 (G2), followed by partial hepatectomy. RESULTS: The results indicated an increase in the number of mitoses after 24 hours and 48 hours (P=0.0022 and P=0.0152, respectively) of PH in the group that received rutin, as well as an increase in AST serum levels after 24 hours (P=0.0159) and 48 hours (P=0.0158) and alkaline phosphatase after 24 hours (P=0.015) in the same group, in relation to the respective controls. The group that received rutin showed a more evident variation than the control group when comparing the 24 hour and 48 hour results regarding AST, number of mitoses and number of apoptosis (P<0.005). CONCLUSION: It was concluded that rutin intervened in hepatic hyperplasia after 24 hours and 48 hours of PH, favoring hepatic hyperplasia.

RESUMO CONTEXTO: A rutina é um flavonoide que pode ser encontrado em grande variedade de vegetais e apresenta atividades anticâncer, anti-inflamatória e antidiabética. OBJETIVO: O objetivo deste estudo foi investigar o efeito da administração oral de rutina sobre a hiperplasia hepática em ratos Wistar submetidos à hepatectomia parcial. MÉTODOS: Foi realizada a análise da hiperplasia hepática e da bioquímica plasmática dos ratos Wistar tratados com rutina 40 mg/kg por 10 dias no grupo 1 (G1) ou salina no grupo 2 (G2), seguido da hepatectomia parcial. RESULTADOS: Os resultados indicaram aumento do número de mitoses após 24 e 48 horas (P=0,0022 e P=0,0152, respectivamente) da hepatectomia parcial no grupo que recebeu rutina, além de um aumento nos níveis séricos de AST após 24 horas (P=0,0159) e 48 horas (P=0,0158) e de fosfatase alcalina após 24 horas (P=0,015) no mesmo grupo, em relação aos respectivos controles. O grupo que recebeu rutina mostrou variação mais evidente do que o grupo controle quando se comparou os resultados de 24 horas e 48 horas em relação a AST, número de mitoses e número de apoptoses (P<0,005). CONCLUSÃO: Foi possível concluir que a rutina interferiu na hiperplasia hepática após 24 e 48 horas após a hepatectomia parcial, favorecendo a hiperplasia hepática.

Quality by design (Qbd) approach to develop stability indicating HPLC method for estimation of rutin in chitosan-sodium alginate nanoparticles

Rutin is a flavonoid glycoside, mainly consists of phenolic compounds, responsible for many biological activities. The objective of the present study was to develop and validate a precise, simple, robust, rapid and reliable reverse phase high -performance liquid chromatography (RP-HPLC) technique by using Qbd approach for evaluating the rutin in nanoparticles. Central composite design (CCD) was employed for optimizing the experimental conditions of RP-HPLC method. Buffer pH, methanol content in the mobile phase composition, flow rate, and wavelength were selected as independent variables whereas retention time, peak area, and asymmetry factor was selected as dependent variables. The retention time, peak area and asymmetric factor of rutin by using optimized independent variables were found to be 3.75 min, 1014.79 mV, and 1.26 respectively. The limit of detection and limit of quantitation values were found to be 0.005 µg/mL and 0.15 µg/mL respectively. For confirming linearity, accuracy, precision, and robustness, the optimized assay condition was validated as per ICH guidelines. The proposed method, which was optimized by QbD approach was found to be a suitable method for analyzing the rutin in chitosan-sodium alginate nanoparticles.

Tomar mate previo a la flebotomía no interfiere en las pruebas bioquímicas de rutina / Drinking mate prior to phlebotomy does not interfere with routine biochemical tests

En este estudio se evaluó el efecto de tomar mate en las pruebas bioquímicas de rutina. Se extrajo sangre a 32 mujeres voluntarias luego de 12 horas de ayuno y a la hora (T1), dos horas (T2) y tres horas (T3) posteriores a la toma de 5 mates. Se estudiaron parámetros hematológicos y analitos de química clínica. Los resultados se analizaron empleando pruebas estadísticas para muestras relacionadas. Se calculó la diferencia porcentual media (DM%) de cada analito en cada hora respecto del valor basal y se comparó con el valor de referencia del cambio (VRC). Una DM% mayor que el VRC se consideró clínicamente significativa. En T1, T2 y T3 los recuentos de neutrófilos, eosinófilos y linfocitos fueron más bajos que en T0, también los niveles de glucosa, urea, creatinina y cistatina C fueron más bajos que en T0, mientras que los valores de proteínas totales, colesterol transportado por lipoproteínas de baja densidad y la actividad enzimática de lactato deshidrogenasa fueron más altos que en T0. En todos los casos los cambios fueron estadísticamente significativos, aunque no lo fueron desde el punto de vista clínico. Tomar 5 mates antes de la flebotomía no interfiere en los resultados de las pruebas bioquímicas de rutina.

In the present study the effect of drinking mate in routine biochemical tests was evaluated. Blood was collected from 32 female volunteers after a 12 h fasting period. In addition, 1 hour (T1), 2 hours (T2), and 3 hours (T3) after drinking 5 mates, blood was collected again. Hematological parameters and clinical chemistry analytes were studied. The results were analyzed using statistical tests for related samples. Mean difference % (MD%) was calculated for each analyte and was further compared with reference change value (RCV). The MDs% higher than RCV were considered clinically significant. At T1, T2, and T3 the count neutrophils, eosinophils and lymphocytes were lower than at T0. Also glucose, urea, creatinine, and cystatin C values were lower than at T0 whereas total proteins, LDL-C, and LD enzymatic activity values were higher than at T0. In all cases, variability was statistically significant but not clinically significant. Drinking 5 mates prior to phlebotomy does not interfere with the results of routine biochemical tests.

Neste trabalho, o efeito de beber chimarrão foi avaliado em testes bioquímicos de rotina. O sangue foi extraído de 32 mulheres voluntárias após 12 horas de jejum, e uma hora (T1), duas horas (T2) e três horas (T3) após a tomada de 5 chimarrões. Parâmetros hematológicos e analitos de química clínica foram estudados. Os resultados foram analisados utilizando testes estatísticos para amostras relacionadas. A diferença percentual média% (DM%) de cada analito em cada hora foi calculada em relação ao valor basal e comparada com o valor de referência da modificação (VRM). Uma DM% maior que o VRM foi considerada clinicamente significativa. Em T1, T2 e T3 as contagens de neutrófilos, eosinófilos e linfócitos foram mais baixas que em T0, Também os níveis de glicose, ureia, creatinina e cistatina C foram mais baixos que em T0, ao passo que os valores de proteínas totais, colesterol transportado por lipoproteínas de baixa densidade e a atividade enzimática de lactato desidrogenase foram mais altos que em T0. Em todos os casos as alterações foram estatisticamente significativas, embora do ponto de vista clínico não o tenham sido. Tomar 5 chimarrões antes da flebotomia não interfere nos resultados dos testes bioquímicos de rotina.

Antidiabetic activity evaluation of Onobrychis species on alloxan-induced diabetic mice

In the current study, four Onobrychis species, O. albiflora Hub.-Mor., O. argyrea Boiss. subsp. argyrea Boiss., O. galegifolia Boiss., and O. tournefortii (Willd.) Desv. were collected from Anatolia to be evaluated for their antidiabetic activities. Methanol water extracts of the aerial parts were used for experiments. An alloxan-induced diabetic mice test model was used. Phytochemical analysis of the tested extracts was investigated using the HPLC method. The highest activity was observed with treatment of O. albiflora aerial part extract. Significant decrements were detected in the blood glucose levels as follows: 180.83±47.48 and 252.83±50.47mg/dL at 100 mg/kg and 200 mg/kg doses of O. albiflora, respectively, when compared to the isotonic saline solution control group, eliciting a blood glucose level of 494.20±27.32. Among the tested standard compounds, rutin and isoquercetin were detected in the examined species. The highest amount of rutin (1.1981±0.0017%) and isoquercetin (0.7318±0.0197%) were found in O. albiflora and O. argyrea subsp. argyrea, respectively. Antidiabetic activities of the tested Onobrychis species seem to indicate a possible correlation with their rutin and isoquercetin contents. Therefore, rutin and isoquercetin may be the antidiabetic compounds that contribute to the antidiabetic activity of the tested Onobrychis species.

Screening for active components of Sophorae Flos on inhibiting AGEs formation based on non-enzymatic glycation reaction / 中国中药杂志

Sugar-poison caused blood-heat is the pathological basis of many complications of diabetes. Advanced glycation end products( AGEs) are considered as the potential glycotoxic factor that can cause blood-heat. Sophorae Flos hold the effect of removing pathogenic heat from blood. In this study,chromatographic non-enzymatic glycation reaction system of bovine serum albumin( BSA)/methylglyoxal( MGO) and Sophorae Flos was established to identify active components in Sophorae Flos inhibiting AGEs formation. The HPLC was used to analyze chromatograms before and after the incubation of Sophorae Flos and methylglyoxal. Changes of chromatographic peaks of eight compounds was found. It is speculated that this change may be due to new substance produced by the reaction of active components in Sophorae Flos and methylglyoxal,and these active components may be flavonoid component rutin. Further investigation for the effects of rutin and MGO reaction( 1 ∶ 1,1 ∶ 3,3 ∶ 1) for 6 days on the formation of AGEs was performed. The results showed that the inhibition activity of rutin on AGEs production was most obvious when the reaction ratio was 1 ∶3,and the most inhibition was in 24 h and stabilized after 3 d. The product of the reaction of rutin with MGO was identified by LC-ESI-MS/MS,which indicated that the newly formed seven substances were the mono-and di-MGO adducts of rutin. This study showed that rutin is the active component on Sophorae Flos for removing pathogenic heat from blood by forming new compounds to inhibit the formation of sugar poison products,which provides reference for rational application of Sophorae Flos.

Efeito da rutina em modelo de depressão crônico induzido por corticosterona em camundongos / Effect of rutin on a model of chronic corticosterone-induced depression in mice

Introdução: a depressão é uma doença altamente prevalente na população mundial e apesar de não ter sua causa estabelecida, algumas teorias tentam esclarecer sua etiologia. A rutina é um flavonoide pertencente à classe dos flavonóis, que tem propriedades anti-inflamatória e antioxidante. Objetivo: avaliar os efeitos antidepressivos da rutina em um modelo crônico induzido por corticosterona em camundongos. Metodologia: foram utilizados camundongos Swiss fêmeas (25-30g) que receberam corticosterona 20mg/kg ou tween 3% por 21dias. Outros grupos receberam corticosterona por 14 dias e entre 15o ao 21o dia de tratamento, rutina (0,2, 2,0 e 10mg/kg) ou fluoxetina10mg/kg ou diazepam 1mg/Kg. Uma hora após a última administração, os animais passaram pelos testes de campo aberto, labirinto em cruz elevada e nado forçado. Após os testes, os animais foram sacrificados por decapitação e as áreas cerebrais córtex pré-frontal, hipocampo e corpo estriado dissecados. Para análise entre os grupos foi usado o teste "t" de Student e para comparação múltipla dos parâmetros utilizará a Análise de Variância (ANOVA). Resultados: a corticosterona foi capaz de induzir a depressão nos animais. No teste de campo aberto, a rutina 0,2, 2,0 e 10mg/kg reduziram a locomoção. A menor dose da rutina apresentou melhor resposta, aumentando o número de entrada e tempo de permanência no braço aberto no teste de labirinto em cruz elevada. Também reduziu significativamente o tempo de imobilização no teste de nado forçado. Conclusão: a substância apresentou atividades ansiolítica e antidepressiva.

Introduction: Depression is a highly prevalent disease in the world population, and, although its cause is not established, several theories try to clarify its etiology. Rutin is a flavonoid belonging to the subclass of flavonoids, which has anti-inflammatory and antioxidant properties. Objective: to evaluate the antidepressant effects of rutin in a chronic model induced by corticosterone in mice. Methods: Female Swiss mice (25-30g) receiving 20mg / kg corticosterone or 3% tween for 21 days were used. Other groups received corticosterone for 14 days and between 15 and 21 days of treatment, rutin (0.2, 2.0 and 10mg / kg) or fluoxetine 10mg / kg or diazepam 1mg / kg. One hour after the last administration, the animals underwent open field tests, elevated cross labyrinth and forced swimming. After the tests, the animals were sacrificed by decapitation and the cerebral areas, prefrontal cortex, hippocampus and striatum dissected. Student's t-test was used for the analysis between the groups, and Variance Analysis (ANOVA) was used for multiple comparison of the parameters. Results: Corticosterone was able to induce depression in animals. In the open field test, routine 0.2, 2.0 and 10mg / kg reduced the locomotion. The lower dose of rutin presented better response, increasing the number of entry and length of stay in the open arm in the high cross maze test. It also significantly reduced stall time in the forced swim test. Conclusion: the substance presented anxiolytic and antidepressant activities.

Analysis of radioprotection and antimutagenic effects of Ilex paraguariensis infusion and its component rutin

DNA repair pathways, cell cycle checkpoints, and redox protection systems are essential factors for securing genomic stability. The aim of the present study was to analyze the effect of Ilex paraguariensis (Ip) infusion and one of its polyphenolic components rutin on cellular and molecular damage induced by ionizing radiation. Ip is a beverage drank by most inhabitants of Argentina, Paraguay, Southern Brazil, and Uruguay. The yeast Saccharomyces cerevisiae (SC7Klys 2-3) was used as the eukaryotic model. Exponentially growing cells were exposed to gamma rays (γ) in the presence or absence of Ip or rutin. The concentrations used simulated those found in the habitual infusion. Surviving fractions, mutation frequency, and DNA double-strand breaks (DSB) were determined after treatments. A significant increase in surviving fractions after gamma irradiation was observed following combined exposure to γ+R, or γ+Ip. Upon these concomitant treatments, mutation and DSB frequency decreased significantly. In the mutant strain deficient in MEC1, a significant increase in γ sensitivity and a low effect of rutin on γ-induced chromosomal fragmentation was observed. Results were interpreted in the framework of a model of interaction between radiation-induced free radicals, DNA repair pathways, and checkpoint controls, where the DNA damage that induced activation of MEC1 nodal point of the network could be modulated by Ip components including rutin. Furthermore, ionizing radiation-induced redox cascades can be interrupted by rutin potential and other protectors contained in Ip.

Desenvolvimento, caracterização e avaliação da formação de complexos de inclusão de rutina, avobenzona e ρ-metoxicinamato de octila na presença de ciclodextrinas / Development, characterization and evaluation of inclusion complexes between rutin, avobenzone and octyl ρ-methoxycinnamate with cyclodextrins

A radiação UV pode causar danos à pele humana, e, evitar estes danos, é uma preocupação crescente para a população e um desafio à comunidade científica. Para uma ação efetiva de fotoproteção, a associação de filtros, como avobenzona (BMBM) e ρ-metoxicinamato de octila (EHMC), são empregados. Devido à semelhança estrutural com os filtros solares químicos, a rutina (RUT), tal como outros flavonoides, apresenta atividade fotoprotetora. Apesar da disponibilidade de diferentes classes de filtros solares, o desenvolvimento de fotoprotetores contendo filtros químicos é um desafio, devido à instabilidade inerente a certos filtros orgânicos. As ciclodextrinas (CDs) são oligossacarídeos cíclicos, de formato tronco-cônico, cuja estrutura externa é hidrófilica e sua cavidade interna central hidrofóbica, com a capacidade de acomodar substâncias lipofílicas, formando complexos de inclusão. A formação dos complexos de inclusão pode levar à alterações de propriedades físico-químicas da molécula hóspede, tais como, solubilidade, fotoestabilidade e biodisponibilidade. O objetivo deste trabalho foi desenvolver, caracterizar e avaliar a formação de complexos de inclusão entre RUT, BMBM e EHMC e as CDs (HPßCD e SBEßCD). Os complexos de inclusão (RUT:HPßCD, RUT:SBEßCD, BMBM:HPßCD, BMBM:SBEßCD, EHMC:HPßCD e EHMC:SBEßCD) foram obtidos pelo método de liofilização e quantificados por cromatografia líquida de alta eficiência (CLAE). Os sistemas binários foram caracterizados em solução, pelo método de equilíbrio de solubilidade, e, no estado sólido, empregando calorimetria exploratória diferencial (DSC), termogravimetria (TG/DTG) e difração de raios-X de pó (PDRX). As substâncias isoladas e os complexos binários foram avaliados quanto à fotoestabilidade em estado sólido, e, em solução. Incremento na solubilidade (X mcg mL-1) foi observado para os complexo RUT:HPßCD (4,13x); RUT:SBEßCD (4,38x); BMBM:HPßCD (43,3x); BMBM:SBEßCD (53,3x); EHMC:HPßCD (12,7x); e EHMC:SBEßCD (70,0x). Os ensaios de DSC, TG/DTG, e P-DRX indicaram a formação de complexos de inclusão para os todos os sistemas, onde a supressão dos eventos endotérmicos característicos das substâncias isoladas foram observados; porém, nos complexos de BMBM, a presença de avobenzona livre no meio foi detectada, sugerindo, que a complexação não foi completa. A formação dos complexos de inclusão promoveu o aumento da fotoestabilidade em todos os sistemas avaliados, tanto no estado sólido, como em solução. Os resultados reportados neste estudo, indicaram que a complexação de substâncias fotoprotetoras com HPßCD e SBEßCD, pode representar, uma estratégia promissora quanto ao aumento da solubilidade e da fotoestabilidade

UV radiation may cause demage on human skin, and preventing it, is an increasing worry for the population and a challenge to the scientific community. For an effective action of photoprotection, the association of filters, like avobention (BMBM) and octyl ρ-methoxycinnamate (EHMC), are used. Due to the structural similarity with the chemical solar filters, the rutin (RUT), like other flavonoids, shows photoprotective activity. Despite the availability of different classes of sunscreens, the development of photoprotectors containing chemical filters is a challenge, due to the inherent instability of certain organic filters. The cyclodextrins (CD) are cyclic oligosaccharides of truncated conical structure, which external structure is hydrophilic and its internal central hydrophobic cavity, with capacity to accommodate lipophilic substances, forming inclusion complexes. The formation of the inclusion complexes can lead to changes in physicalchemical properties of the host molecule, such as, solubility, photostability and bioavailability. The objective of this work was to develop, characterize and evaluate the formation of the inclusion complexes between RUT, BMBM and EHMC and the CDs (HPßCD and SBEßCD). The inclusion complexes (RUT:HPßCD, RUT:SBEßCD, BMBM:HPßCD, BMBM:SBEßCD, EHMC:HPßCD and EHMC:SBEßCD) were obtained by the lyophilization method and quantified by high performance liquid chromatography (HPLC). The binary systems were characterized in solution, by solubility equilibrium method and in solid state, using differential scanning calorimetry (DSC), thermogravimetry (TG/DTG) and powder X-ray diffraction (P-XRD). The isolated substances and binary complexes were evaluated the photostability in solid state, and in solution. The increase in solubility (X mcg mL-1) was observed for the complexes RUT:HPßCD (4.13x); RUT:SBEßCD (4.38x); BMBM:HPßCD (43.3x); BMBM:SBEßCD (53.3x); EHMC:HPßCD (12.7x); and EHMC:SBEßCD (70.0x). The analysis of DSC, TG/DTG, and P-DRX indicated the formation of inclusion complexes for all systems, where the suppression of the endothermic events characteristic of the isolated substances were observed; however, in the BMBM complexes, the presence of free avobenzone was detected, suggesting that the complexation was not complete. The formation of inclusion complexes promoted the increase of photostability in all evaluated systems, as in solid state as in solution. The results reported in this study indicated that the complexation of photoprotective substances with (HPßCD e SBEßCD). may represent a promising strategy for increasing solubility and photostability

Influence of rutin on the effects of neonatal cigarette smoke exposure-induced exacerbated MMP-9 expression, Th17 cytokines and NF-κB/iNOS-mediated inflammatory responses in asthmatic mice model

Allergic asthma is one of the most enduring diseases of the airway. The T-helper cells and regulatory T-cells are critically involved in inflammatory responses, mucus hypersecretion, airway remodelling and in airway hyper-responsiveness. Cigarette smoke (CS) has been found to aggravate inflammatory responses in asthma. Though currently employed drugs are effective, associated side effects demand identification and development of novel drugs with negligible or no adverse effects. Rutin, plant-derived flavonoid has been found to possess antioxidant and anti-inflammatory effects. We investigated the ability of rutin to modulate T-cells and inhibit inflammation in experimentally-induced asthma in cigarette smoke exposed mice. Separate groups of neonatal mice were exposed to CS for 10 days from post-natal days 2 to 11. After 2 weeks, the mice were sensitized and challenged with ovalbumin (OVA). Treatment group were given rutin (37.5 or 75 mg/kg body weight) during OVA sensitization and challenge. Rutin treatment was found to significantly inhibit cellular infiltration in the airways and Th2 and Th17 cytokine levels as well. Flow cytometry revealed effectively raised CD4⁺CD25⁺Fox3⁺ Treg cells and supressed Th17 cell population on rutin treatment. Airway hyper-responsiveness observed following CS and OVA challenge were inhibited by rutin. NF-κB and iNOS, chief regulators of inflammatory responses robustly activated by CS and OVA were down-regulated by rutin. Rutin also inhibited the expression of matrix metalloproteinase 9, thereby aiding in prevention of airway remodelling in asthma thereby revealing to be a potent candidate in asthma therapy.

Identification and HPLC Quantification of a Phytoecdysone and Three Phenolic Glycosides in Lamium takesimense Nakai

The herbs of Lamium takesimense Nakai (Lamiaceae) is used to treat spasmodic and inflammatory disease. The four polar compounds, ecdysterone, isoacteoside, rutin and lamiuside C, were isolated and identified from the BuOH fraction of the L. takesimense MeOH extract. HPLC quantification was performed on a Capcell Pak C18 column (5 µm, 4.6 mm × 250 mm) with a gradient elution of H₂O and 0.05% acetic acid in MeOH. The HPLC method was validated in terms of linearity, sensitivity, stability, precision, and accuracy. The quantitative level in plant material was determined as the following order: lamiuside C (4, 3.75 mg/g dry weight) > ecdysterone (1, 1.93 mg/g) > isoacteoside (2, 1.32 mg/g) > rutin (3, 0.97 mg/g).

Content Analysis of Rutin in the Leaves of Boehmeria nivea Harvested in Different Regions of South Korea by HPLC-UV

Phytochemical analysis of Boehmeria nivea (Bn) leaves by medium pressure liquid chromatography led to the isolation of a flavonoid glycoside identified by spectroscopic analysis as rutin. The amount of rutin in the leaves of Bn harvested from nine regions in South Korea (Bn 1–9) which were collected on the months of June, July, August, and September was determined by HPLC-UV analysis. A gradient elution program that utilizes a Discovery® C18 (4.6 × 250 mm, 5 µm) column and mobile phase composed of 1% acetic acid-water: acetonitrile (90:10 to 60:40 for min) was followed. The injection volume and flow rate were 10 µl and 1 mL/ min, respectively. UV detection was set at 350 nm. Results show that Bn-8 harvested in September reported the highest content of rutin among the samples analyzed. This study provides a basis for the optimal harvest time of Bn which maximizes the yield of rutin.

Efeitos pós-ingestivos de flavonóides no epitélio do intestino médio de larvas de Anticarsia gemmatalis (Hübner 1818) (Lepidoptera: Erebidae) / Post-ingestive effects of flavonoids in the midgut epithelium of Anticarsia gemmatalis (Hübner 1818) (Lepidoptera: Erebidae) larvae

An alternative control to soybean caterpillar has been the use of insect resistant plants that present phenolic flavonoids. The midgut is the main access pathway of food and chemical substances ingested. The present study examined morphological alterations in the midgut epithelium of the soybean caterpillar, after the ingestion of soybean genotypes containing the flavonoids rutin and genistin. The caterpillars and genotypes (BRS 257 - control, BR 16, Dowling, PI 229358, IAC 100 and PI 227687) were obtained from the insect rearing facility at the Embrapa Soja. The midguts were collected, fixed in Karnovsky, processed and analyzed under a light microscope. All treatments caused alterations in the midgut epithelial cells. These alterations in the columnar cells were more assiduous than the on other cell types, showing an increase of cytoplasmic protrusions and vacuoles. The goblet cells showed few changes for all genotypes tested, while the regenerative cells presented alterations mainly for the Dowling and PI 227687 treatments. The peritrophic membrane was absent for genotypes IAC100 and PI 227687. We conclude that the Dowling and PI 227687 genotypes were effective and promoted great morphological alterations in the midgut of the soybean caterpillars, being able to be very effective for the control of this plague.

Um controle alternativo para a lagarta da soja tem sido o uso de plantas resistentes a insetos que contém flavonoides fenólicos. O intestino médio é a principal via de acesso do alimento e substâncias químicas ingeridas. O presente estudo examinou as alterações morfológicas no epitélio do intestino médio da lagarta da soja, após a ingestão de genótipos de soja contendo os flavonoides rutina e genistina. As lagartas e os genótipos (BRS 257 - controle, BR 16, Dowling, PI 229358, IAC 100 e PI 227687) foram obtidos do laboratório de criação de insetos da Embrapa Soja. Os intestinos médios foram coletados, fixados em Karnovsky, processados e analisados ao microscópio de luz. Todos os tratamentos causaram alterações nas células epiteliais do intestino médio. As alterações foram mais assíduas nas células colunares do que nos demais tipos celulares. Essas apresentaram grande quantidade de protrusões citoplasmáticas e de vacúolos. As células caliciformes apresentaram poucas alterações para todos os genótipos testados, enquanto as células regenerativas apresentaram alterações principalmente nos tratamentos Dowling e PI 227687. A membrana peritrófica estava ausente para os genótipos IAC100 e PI 227687. Conclui-se que os genótipos Dowling e PI 227687 foram bastante efetivos e promoveram grandes alterações morfológicas no intestino médio das lagartas da soja, podendo ser bastante eficaz para o controle desta praga.