Composición química y caracterización de flavonoides de extractos metanólicos de hojas de dos tipos de Ruta chalepensis L / Chemical composition and flavonoids characterization of the methanolic extract of leaves of two types of Ruta Chalepensis L

Objetivos. Detectar metabolitos secundarios y caracterizar estructuras químicas de flavonoides de los extractos metanólicos de hojas de dos tipos de Ruta chalepensis L. «ruda macho¼ y «ruda hembra¼. Materiales y métodos. Se elaboraron extractos metanólicos de hojas de Ruta chalepensis L. «ruda macho¼ y «ruda hembra¼. Se llevó a cabo la prueba de solubilidad de los extractos obtenidos utilizando solventes de polaridad creciente. Posteriormente, se detectaron los metabolitos secundarios presentes en los extractos mediante la ejecución del tamizaje fitoquímico, donde se utilizaron diversos reactivos Shinoda, Dragendorff, Borntrager, entre otros. Se utilizó el método de cromatografía en capa fina, espectrofotometría UV/Vis y reactivos de desplazamiento para caracterizar las estructuras químicas de los flavonoides presentes en los extractos metanólicos de hojas de Ruta chalepensis L. «ruda macho¼ y «ruda hembra¼. Resultados. El extracto metanólico de hojas de Ruta chalepensis L. «ruda macho¼ presentó una mayor afinidad por solventes polares, mientras que en el extracto metanólico de hojas de Ruta chalepensis L. «ruda hembra¼ fue soluble en solventes medianamente polares. Se detectaron metabolitos secundarios tales como: taninos, alcaloides y flavonoides en ambos tipos. Por otro lado, se caracterizaron diez estructuras químicas tipo flavonoides a través del análisis de los espectros UV/Vis y utilizando reactivos de desplazamiento, de las cuales cinco corresponden a Ruta chalepensis L. «ruda macho¼ y las restantes a Ruta chalepensis L. «ruda hembra¼. Conclusiones. Se detectaron algunos metabolitos secundarios caracterizándose diez estructuras químicas de flavonoides en los extractos metanólicos de hojas de Ruta chalepensis L. «ruda macho¼ y «ruda hembra¼. Asimismo, la presencia de rutina en «ruda hembra¼ es la principal característica que la diferencia de «ruda macho¼.

Objective. To detect the secondary metabolites and characterize the chemical structures of the flavonoids in the methanolic extracts of leaves of the Ruta chalepensis L. "ruda macho" and "ruda hembra" types. Materials and methods. We elaborate the methanolic extracts of Ruta chalepensis L. leaves "ruda macho" and "ruda hembra". Then, the solubility test of the obtained extracts was carried out using solvents of increasing polarity. Subsequently, the secondary metabolites present in the extracts were detected by executing the phytochemical screening, various reagents were used: Shinoda, Dragendorff, Borntrager, among others. The method of thin layer chromatography, UV / Vis spectrophotometry and displacement reagents was used to characterize the chemical structures of the flavonoids in the methanolic extracts of leaves of the Ruta chalepensis L. "ruda macho" and "ruda hembra". Results. The methanolic extract of Ruta chalepensis L. "ruda macho" leaves showed a greater affinity for polar solvents, while the methanolic extract of Ruta chalepensis L. "ruda hembra" leaves was soluble in moderately polar solvents. Secondary metabolites were detected, such as: tannins, alkaloids and flavonoids in both types. On the other hand, ten flavonoidtype chemical structures were characterized through the analysis of UV / Vis spectra and using displacement reagent, of which five of them correspond to Ruta chalepensis L. "ruda macho" and the others to Ruta chalepensis L. "ruda hembra". Conclusions. Some secondary metabolites were detected and ten chemical structures of flavonoids were characterized in the methanolic extracts of Ruta chalepensis L. "ruda macho" and "ruda hembra" leaves. Likewise, the presence of rutina in "ruda hembra" is the main characteristic that differentiates it from "ruda macho".

Relaxant effects of a hydroalcoholic extract of Ruta graveolens on isolated rat tracheal rings

BACKGROUND: Ruta graveolens L. (R. graveolens) is a medicinal plant employed in non-traditional medicines that has various therapeutic properties, including anthelmintic, and vasodilatory actions, among others. We evaluated the trachea-relaxant effects of hydroalcoholic extract of R. graveolens against potassium chloride (KCl)- and carbachol-induced contraction of rat tracheal rings in an isolated organ bath. RESULTS: The results showed that the airway smooth muscle contraction induced by the depolarizing agent (KCl) and cholinergic agonist (carbachol) was markedly reduced by R. graveolens in a concentration-dependent manner, with maximum values of 109 ± 7.9 % and 118 ± 2.6 %, respectively (changes in tension expressed as positive percentages of change in proportion to maximum contraction), at the concentration of 45 µg/mL (half-maximal inhibitory concentration IC50: 35.5 µg/mL and 27.8 µg/mL for KCl- and carbachol-induced contraction, respectively). Additionally, the presence of R. graveolens produced rightward parallel displacement of carbachol dose-response curves and reduced over 35 % of the maximum smooth muscle contraction. CONCLUSIONS: The hydroalcoholic extract of R. graveolens exhibited relaxant activity on rat tracheal rings. The results suggest that the trachea-relaxant effect is mediated by a non-competitive antagonistic mechanism. More detailed studies are needed to identify the target of the inhibition, and to determine more precisely the pharmacological mechanisms involved in the observed biological effects.

Effects of the chronic ingestion of an infusion of Ruta chalepensis on the vasomotor responses of rat aortic rings / Efectos de la ingestión crónica de una infusión de Ruta chalepensis sobre las respuestas vasomotoras de anillos de aorta de rata

Ruta chalepensis, is used, in traditional medicine, as emmenagogue, abortive, and analgesic. We analyzed, in male Wistar rats, the effects of the chronic intake of an infusion of Ruta chalepensis (20 g/L) on the vasomotor responses of, either intact or endothelium-denuded aortic rings, to phenylephrine or carbachol. Only in rings with endothelium significant effects were observed. The infusion induced a leftward shift of the concentration-response curve to phenylephrine and an increase in maximal tension development. These effects were abolished by indomethacin. In these rings, inhibiting the synthesis of nitric oxide, in the presence of indomethacin, induced a leftward shift of the concentration response curve to phenylephrine, as well as an increase in maximal tension. These results suggest that the chronic ingestion of a Ruta chalepensis infusion induces an endothelium dependent increase in the synthesis/release of cyclooxygenase-dependent vasoconstrictor prostanoids, and an increase in the basal synthesis/release of nitric oxide.

Ruta chalepensis se utiliza en la medicina tradicional como emenagogo, abortivo y analgésico. Se analizaron, en ratas Wistar macho, los efectos de la ingesta crónica de una infusión de Ruta chalepensis (20 g /L), sobre las respuestas vasomotoras de anillos de aorta con y sin endotelio, a la fenilefrina o al carbacol Se observaron efectos significativos sólo en anillos con endotelio. La infusión indujo un desplazamiento a la izquierda de la curva de concentración-respuesta a fenilefrina y un incremento en la tensión máxima desarrollada. Estos efectos fueron abolidos por la indometacina. La inhibición de la síntesis de óxido nítrico, en presencia de indometacina, produjo un desplazamiento a la izquierda de la curva de concentración-respuesta a la fenilefrina, así como un incremento en la tensión máxima. Estos resultados sugieren que la ingesta crónica de una infusión de Ruta chalepensis induce un incremento en la síntesis/liberación de prostanoides vasoconstrictores dependientes de la ciclooxigenasa y un aumento en la síntesis /liberación basal de óxido nítrico.

Fagodisuasión de un extracto de ruda (Ruta chalepensis, Rutaceae) y sus particiones sobre larvas de Hypsipyla grandella (Lepidoptera: Pyralidae)

Phagodeterrence by a crude extract of common rue (Ruta chalepensis, Rutaceae) and its partitions on Hypsipyla grandella (Lepidoptera: Pyralidae) larvae. Hypsipyla grandella (Zeller) larva is maybe the main forest pest in Latin America and the Caribbean, as it bores into the main shoot of trees providing precious woods, such as mahoganies (Swietenia spp.) and cedars (Cedrela spp.). In the search for a preventative method for managing it, the crude leaf extract of common rue (Ruta chalepensis L.), as well as four partitions thereof (water, hexane, dichlorometane, and ethyl acetate), were tested for phagodeterrence. Laboratory bioassays involved increasing concentrations of the crude extract (0.1, 0.32, 1.0, 3.20 and 10.0%w/v) as well as each one of the partitions (in accordance to the yield obtained from the partitioning process), plus rutin, a flavone glycosid. A randomized complete block design, with four replicates, was used. H. grandella instar III larvae were exposed for 24 h to cedar (Cedrela odorata) leaf discs dipped into the respective treatment, after which disc consumption was measured. Strong phagodeterrence was detected at concentrations as low as 0.32 and 0.074%w/v for the crude extract and the hexane partitions, respectively; the ethyl acetate (0.24%w/v) and the water partition (for all of its concentrations), as well as the rutin (starting at 0.064%w/v), caused phagodeterrence, too. Moreover, the crude extract was submitted to a phytochemical screening by means of a number of qualitative tests, to determine possible metabolites causing phagodeterrence, the most important being alkaloids, triterpenes, coumarins and rutin. A particular phytochemical screening was carried out for the hexane partition, which was the most active. Rev. Biol. Trop. 58 (1): 1-14. Epub 2010 March 01.

La larva de Hypsipyla grandella (Zeller) es quizás la principal plaga forestal en América Latina y el Caribe, por perforar el brote principal de árboles de maderas reciosas, como caobas (Swietenia spp.) y cedros (Cedrela spp.). En la búsqueda de un método preventivo para su manejo, se estudió la actividad fagodisuasiva del extracto crudo y de cuatro particiones fitoquímicas (agua, hexano, diclorometano y acetato de etilo) del follaje de ruda (Ruta chalepensis L.). Se realizaron bioensayos de laboratorio con concentraciones crecientes del extracto crudo (0.1; 0.32; 1.0; 3.20 y 10.0%m/v) y con cada una de las particiones (según el rendimiento del proceso de particionamiento), así como con el glicósido flavónico rutina. Para ello se empleó un diseño de bloques completamente al azar, con cuatro repeticiones y se expusieron larvas de instar III de H. grandella a discos de cedro (Cedrela odorata) impregnados con el respectivo tratamiento, por 24h. Se midió el porcentaje de consumo de cada disco. Hubo un fuerte efecto fagodisuasivo en el extracto crudo y en la partición hexano, a concentraciones tan bajas como 0.32% y 0.074% m/v, respectivamente; las particiones de acetato de etilo (0.24%m/v) y la acuosa (en todas sus concentraciones), así como la rutina (a partir del 0.064%m/v) también causaron fagodisuasión. Además, se realizó un tamizaje fitoquímico del extracto crudo, utilizando varias pruebas cualitativas para detectar la presencia de los principales grupos de metabolitos secundarios, entre los cuales los más importantes fueron alcaloides, triterpenos, cumarinas y la rutina. Asimismo, se realizó una marcha fitoquímica a la partición hexano, por ser la más activa.

Actividad fagodisuasiva y sistémica de una formulación derivada de un extracto de ruda (Ruta chalepensis, Rutaceae) sobre larvas de Hypsipyla grandella (Lepidoptera: Pyralidae)

Phagodeterrent and systemic activity of a fomulation derived from an extract of common rue (Ruta chalepensis, Rutaceae) on Hypsipyla grandella (Lepidoptera: Pyralidae) larvae. A key neotropical pest of mahoganies (Swietenia spp.) and cedars (Cedrela spp.), the larva of Hypsipyla grandella (Zeller) commonly bores into the main shoot of these valuable timber species, causing forking of the stems. Therefore, it would be desirable to count upon a phagodeterrent and systemic product which could readily protect young tissues. Since crude extracts of common rue foliage (Ruta chalepensis L.) have phagodeterrent activity on H. grandella larva, a product was formulated by combining such an extract with a number of coadjuvants, and was assessed on 75-100cm tall Spanish cedar (Cedrela odorata L.) trees, inside a greenhouse. A completely randomized design was used, with 10 trees per each one of the following treatments: the formulated product, an absolute control (distilled water), a relative control (carbofuran, 1%m/v in water), and the relative control without the crude extract. Number of attacks to the main shoot, fallen leaves and frass piles were recorded, as well as tunnel length. Regarding these variables, the formulated product provided excellent protection to treated trees, with only 0.1 attacks to the main shoot, on the average, at the end of the evaluation period. Likewise, when the systemic activity of the crude extract was assessed, substances present in it were able to translocate and get assimilated by the trees, thereafter causing phagodeterrence to larvae. Finally, an espectrophotometry analysis (performed at a wavelength of 355nm) allowed us to determine the concentration (403±1mg/l) and percentage (0.40%) of the flavonoid rutin in the crude extract, which eventually may be used as an analytical marker compound, should an industrial formulation be developed. Rev. Biol. Trop. 58 (1): 15-29. Epub 2010 March 01.

Para el manejo de Hypsipyla grandella (Zeller), cuya larva barrena el brote principal de caobas (Swietenia spp.) y cedros (Cedrela spp.), sería deseable contar con un producto fagodisuasivo y con buena actividad sistémica, para que proteja rápidamente los tejidos jóvenes. Por tanto, conociendo que un extracto crudo del follaje de ruda (Ruta chalepensis L.) posee actividad fagodisuasiva contra dicho insecto, se formuló un producto combinando dicho extracto con varios coadyuvantes, y se evaluó en árboles de cedro amargo (Cedrela odorata L.) de 75- 100cm de altura, en un invernadero. Se utilizó un diseño completamente al azar, con 10 árboles por cada tratamiento, los que correspondieron al producto formulado, a un testigo absoluto (agua destilada), un testigo relativo (carbofurán al 1%m/v en agua) y el testigo relativo a la formulación sin el extracto crudo. Las variables medidas fueron los ataques al brote principal, el número de hojas caídas, el número de montículos y la longitud de los túneles. Para dichas variables, el producto formulado aportó excelente protección a los árboles tratados, con apenas 0.1 ataques al brote principal, en promedio, al final del período de evaluación. Asimismo, al evaluar la actividad sistémica del extracto, las sustancias presentes en él pudieron desplazarse y ser asimiladas por los árboles, causando fagodisuasión en las larvas. Finalmente, el análisis espectrofotométrico (a una longitud de onda de 355nm) permitió determinar la concentración (403±1mg/l) y el porcentaje (0.40%) del flavonoide rutina en el extracto crudo, lo cual eventualmente podría utilizarse como un compuesto marcador analítico, si llegara a desarrollarse una formulación industrial.

Inhibition of chemically induced carcinogenesis by drugs used in homeopathic medicine.

Homeopathy is considered as one modality for cancer therapy. However, there are only very few clinical reports on the activity of the drugs, as well as in experimental animals. Presently we have evaluated the inhibitory effects of potentized homeopathic preparations against N'-nitrosodiethylamine (NDEA) induced hepatocellular carcinoma in rats as well as 3-methylcholanthrene-induced sarcomas in mice. We have used Ruta, Hydrastis, Lycopodium and Thuja, which are commonly employed in homeopathy for treating cancer. Administration of NDEA in rats resulted in tumor induction in the liver and elevated marker enzymes such as gamma-glutamyl transpeptidase, glutamate pyruvate transaminase, glutamate oxaloacetate transaminase and alkaline phosphatase in the serum and in liver. Concomitant administration of homeopathic drugs retarded the tumor growth and significantly reduced the elevated marker enzymes level as revealed by morphological, biochemical and histopathological evaluation. Out of the four drugs studied, Ruta 200c showed maximum inhibition of liver tumor development. Ruta 200c and phosphorus 1M were found to reduce the incidence of 3-methylcholanthrene-induced sarcomas and also increase the life span of mice harboring the tumours. These studies demonstrate that homeopathic drugs, at ultra low doses, may be able to decrease tumor induction by carcinogen administration. At present we do not know the mechanisms of action of these drugs useful against carcinogenesis.

Anti-tumour activity of Ruta graveolens extract.

An extract of Ruta graveolens was found to be cytotoxic to Dalton's lymphoma ascites (DLA), Ehrlich ascites carcinoma (EAC) and L929 cells in culture (IC100=16 mg/ml) and also to increase the lifespan of tumour bearing animals. The extract further decreased solid tumours developing from DLA and EAC cells when given simultaneously with elongation of the lifespan of tumour-bearing animals. A homeopathic preparation of Ruta graveolens (200 c) was equally effective. Neither was effective for reducing already developed tumours. The Ruta graveolens extract was found to scavenge hydroxyl radicals and inhibit lipid peroxidation at low concentrations. However, at higher concentrations the extract acted as a prooxidant as inhibition of lipid peroxidation and scavenging of hydroxyl radical was minimal. These data indicates that the prooxidant activity of Ruta graveolens may be responsible for the cytocidal action of the extract and its ability to produce tumour reduction.

Effect of Ruta graveolens L. and Euphorbia peplus L. anti-inflammatory extracts on nutritional status of rats and the safety of their use.

A significant increase in body weight with remarkable increase in total food intake and significant increase in protein efficiency ratio were observed following oral administration of R. graveolens ether extract (500 mg/kg body wt) to growing rats for 3 weeks. Serum albumin was significantly decreased after administration of declofenac (15 mg/kg body wt). Albumin/globulin ratio decreased significantly on administration of E. peplus ether extract (500 mg/kg body wt). No significant changes were observed in other biochemical and nutritional parameters on administration of either of the extracts or declofenac. However, only a significant elevation of alkaline phosphatase was noticed during treatment with R. graveolens. The results suggest that both plant extracts have no harmful effect on nutritional status and are safe towards kidney functions, while Euphorbia is more safe than Ruta in relation to liver functions.