Preclinical studies of hydroalcoholic extract of Calea uniflora Less / Estudios preclínicos de extracto hidroalcohólico de Calea uniflora Less

Calea uniflora Less known popularly as Arnica in Brazil, is a native plant from Brazil, popular used by coastal populations from south of Santa Catarina. The purpose of this study was to verify the safety profile in of hydroalcoholic extract of C. uniflora in florescences.The hydroalcoholic extract of C. uniflora in florescences was evaluated for its acute and sub-acute toxicity. Acute topical toxicity was performed using the methodology of guideline 402 from OECD. Acute oral toxicity was performed using the methodology of guideline 423 from OECD and sub-acute toxicity was performed using the methodology adapted of guideline 407 from OECD. The single dose for oral or topical administration of C. uniflora showed DL50> 5000 mg/kg b.w. The sub-acute treatment induced animal death in groups, which was administered extract in the doses 100, 250, 500 and 1000 mg/kg. The main signs of toxicity observed were respiratory difficulty, increase in lung weigh, lung damage and muscular relation. The topical or oral administration of C. uniflora extract in short period did not caused toxicological effects in animals, however, when administered for a longer period and in concentrations of 250, 500 and 1000 mg/kg (oral.) caused lung damage and even the death of the animal.

Calea uniflora Less conocida popularmente como Arnica en Brasil, es una planta nativa de Brasil, popularmente utilizada por poblaciones costeras del sur de Santa Catarina. El objetivo de este estudio fue verificar el perfil de seguridad del extracto hidroalcoólico de inflorescencias de C. uniflora. El extracto hidroalcoólico de inflorescencias de C. uniflora fue evaluado en cuanto a su toxicidad aguda y subaguda. La toxicidad tópica aguda se realizó utilizando la metodología de la directriz 402 de la OECD. La toxicidad oral aguda fue realizada usando la metodología de la directriz 423 de la OECD y la toxicidad subaguda fue realizada usando la metodología adaptada de la directriz 407 de la OECD. La dosis única para administración oral o tópica de C. uniflora mostro DL50> 5000 mg/kg. El tratamiento subagudo indujo la muerte de animales en grupos a los que se administró extracto en las dosis de 100, 250, 500 y 1000 mg/kg. Los principales signos de toxicidad observados fueron dificultad respiratoria, aumento del peso del pulmón, daño pulmonar y relación muscular. La administración tópica oral del extracto de C. uniflora a corto plazo no causó efectos toxicológicos en los animales, mientras que, cuando se administró por un período mayor y en las concentraciones de 250, 500 y 1000 mg/kg (oral) causaron danos en los pulmones y hasta la muerte del animal.

Chromenes from leaves of Calea pinnatifida and evaluation of their leishmanicidal activity

Calea pinnatifida (R. Br.) Less., Asteraceae, is popularly known as “quebra-tudo”, “cipó-cruz” or “aruca”. This species is used in the folk medicine for the treatment of stomach pain, giardiasis and amoebiasis. The aim of this study was to isolate and identify chromenes from leaves of C. pinnatifida and evaluate their leishmanicidal activity. A fraction from leaves of C. pinnatifida was analyzed for their chemical constituents, resulting in the isolation and characterization of four known chromenes: 6-acetyl-7-hydroxy-2,2-dimethylchromene (1), 6-acetyl-7-methoxy-2,2-dimethylchromene (2), 6-(1-hydroxyethyl)-7-methoxy-2,2-dimethylchromene (3) and 6-(1-ethoxyethyl)-7-methoxy-2,2-dimethylchromene (4). Structure identification of isolated compounds involved analysis of spectral data of 1D and 2D-NMR. The isolated compounds are here reported for the first time in C. pinnatifida, and the chromenes 1 and 3 show a moderate leishmanicidal activity.

Anatomical Characters of Leave and Stem of Calea serrata Less., Asteraceae

The genus Calea belongs to the tribe Heliantheae and presents about 125 species. Calea serrata, popularly known as erva-de-cobra, chá-amargo and quebra-tudo, is an endemic species found in southern Brazil and is used in traditional medicine to treat ulcers and livers problems. The present work aimed to study the pharmacobotanical characters of leaves and stems of C. serrata for quality control purposes. The plant material was processed according to standard methods of light and scanning electron microscopy. Glandular capitate-stalked and capitate-sessile, uniseriate multicellular non-glandular trichome with tapered apical cell, conical non-glandular trichome, isobilateral mesophyll, secretory ducts near the endoderm and circular shape with six ribs in the stem were important characters, which contributed to the identification of the species.

Effects of the butanol-containing fraction from leaves of Calea prunifolia H.B.K. on blood pressure and smooth muscle responses of Wistar rats / Efecto de la fracción butanólica de las hojas de Calea prunifolia H.B.K. sobre la presión arterial y la relajación muscular en ratas wistar

The butanol-containing fraction from leaves of Calea prunifolia H.B.K. was obtained in order to examine cardiovascular effects on two distinct rat preparations. The responses elicited in isolated aorta and isolated vas deferens, were examined, as were the changes in blood pressure in anesthetized and conscious Wistar rats. In addition, the effects on angiotensin-converting enzyme activity in plasma and the effects on cytosolic calcium in cardiomyocytes and uterine cells were measured. Results show that the butanol-containing fraction from C. prunifolia (0.1-100 µg/mL) relaxes phenylephrine-induced contractions in isolated aorta (CI50: 58 µg/mL), decreases the contractile response induced by noradrenaline in vas deferens, and shows a competitive antagonism profile (pA2: 4.48, m: 1024). The butanol-containing fraction from C. prunifolia also decreases blood pressure in anesthetized rats in a dose-dependent manner (1-100 mg/kg, i.v.), yet is devoid of hypotensive effects in normotensive, conscious rats. In addition, C. prunifolia does not modify the hypertensive effects induced by the nitric oxide synthase inhibitor L-NAME, and it also does not affect the angiotensin-converting enzyme plasma activity nor the cytosolic calcium in cardiomyocytes and uterine cells. According to these results, C. prunifolia relaxes smooth muscle and vascular tone, favoring the decrease in blood pressure in rats via mechanisms related to alpha adrenergic inhibition.

Se evaluó el efecto cardiovascular en ratas inducido por la fracción butanólica de las hojas de Calea prunifolia H.B.K., examinando el efecto generado en preparaciones de anillos aislados de aorta y conducto deferente, y los cambios de presión arterial en ratas wistar anestesiadas y despiertas. También se cuantificó el efecto sobre la enzima convertidora de angiotensina (ECA) en plasma y sobre el calcio citosólico en cardiomiocitos y uteromiocitos. Los resultados mostraron que la fracción butanólica de C. prunifolia (0,1-100 µg/mL) relajaba anillos de aorta contraídos con fenilefrina (CI50: 58 µg/mL) y disminuía la contracción del conducto deferente inducido por noradrenalina con un perfil de inhibición competitiva (pA2: 4.48, m: 1.024). La fracción butanólica de C. prunifolia disminuye la presión sanguínea en ratas anestesiadas en función de la dosis (1-100 mg/kg, i.v.), si bien estuvo desprovista de efectos hipotensores en ratas despiertas. C. prunifolia tampoco modificó el efecto hipertensor inducido por el inhibidor de óxido nítrico sintetasa L-NAME, ni los niveles de ECA, ni la concentración de calcio citosólico en cardio y uteromiocitos. De acuerdo con estos resultados, C. prunifolia relaja el músculo liso y disminuye el tono vascular favoreciendo la disminución de la presión arterial en ratas por medio de mecanismos vinculados con la inhibición alfa adrenérgica.

ACTIVIDAD ANTIINFLAMATORIA DE EXTRACTOS Y FRACCIONES DE Myrcianthes leucoxila, Calea prunifolia, Curatella americana Y Physalis peruviana EN LOS MODELOS EDEMA AURICULAR POR TPA, EDEMA PLANTAR POR CARRAGENINA Y ARTRITIS INDUCIDA POR COLÁGENO / ANTIINFLAMMATORY ACTIVITY OF EXTRACTS AND FRACTIONS OF Myrcianthes leucoxila, Calea prunifolia, Curatella americana Y Physalis peruviana OBTAINED FROM ON TPA-INDUCED EAR OEDEMA, CARRAGEENANINDUCED PAW OEDEMA AND COLLAGEN-INDUCED ARTHRITIS

Introducción. El empleo etnofarmacológico de plantas en el manejo de procesos inflamatorios crónicos y la necesidad su caracterización farmacológica, promueven la evaluación de actividad antiinflamatoria de sustancias en modelos in vivo. Materiales y métodos. Evaluación de extractos y fracciones de Calea prunifolia (CP), Curatella americana (CA), Myrcianthes leucoxila (ML) y Physalis peruviana (PP) sobre los modelos edema auricular por acetato de tetradecanoilforbol (TPA) en ratón albino ICR y edema plantar por carragenina en ratas Wistar, seleccionando un extracto para valorar su actividad antiartrítica en el modelo artritis inducida por colágeno en ratones DBA. Resultados. Las fracciones con mayor porcentaje de inhibición del edema en el modelo edema auricular por TPA fueron ML etanólica total (82±6%), CP rica en terpenos (81±6%) y CA rica en terpenos (81±7%) (P<0,05). No se obtuvo actividad antiinflamatoria significativa sobre el modelo edema plantar por carragenina. Se evaluó la actividad antiartrítica de la fracción rica en terpenos de ML sobre el modelo artritis inducida por colágeno, sin encontrarse efecto significativo sobre edema de patas traseras, peso corporal, escala histopatológica de severidad de artritis ni inmunohistoquímica para factor de necrosis tumoral alfa (P>0,05). Discusión. La actividad antiinflamatoria en el modelo de inflamación aguda edema auricular por TPA para los extractos y fracciones de CP, CA y ML se puede relacionar con la afectación de mediadores relacionados con fosfolipasa A2 dado el nivel de efecto similar a indometacina encontrado. La fracción terpénica de ML no mostró actividad antiartrítica ni modificó la expresión de TNF-a en el modelo de artritis crónica autoinmune empleado, por lo cual no posee actividad inmunomoduladora ni antiinflamatoria en la dosis evaluada. Conclusión: Las fracciones terpénicas de los extractos de CA y CP y los extractos metanólicos de ML mostraron una actividad antiinflamatoria significativa en el edema auricular inducida por TPA. Estos extractos tuvieron poca actividad sobre el edema inducido por carragenina en la pata. La fracción terpénica del extracto ML no presentó actividad antiartrítica en el modelo de artritis inducido por el colágeno.

Introduction. Ethnopharmacological use of plants in management of chronic inflammatory diseases and the need to have their pharmacological characterization promote the evaluation of anti-inflammatory activity over in vivo models. Materials and methods. Evaluation of extracts and fractions of Calea prunifolia (CP), Curatella americana (CA), Myrcianthes leucoxila(ML) and Physalis peruviana (PP) on auricular edema by tetradecanoylphorbol acetate (TPA)-in ICR albino mice and carrageenan-induced leg edema in Wistar rats selecting an extract to evaluate its anti-arthritic activity in collagen-induced arthritis model in DBA mice. Results. The fractions with greater edema inhibition percentage on TPA-induced ear edema included whole ethanolic fraction of ML (82±6%), CP terpenes rich fraction (81±6%) and CA terpenes rich fraction (81±7%) (P<0.05). Significant antiinflamatory activity was not obtained on the carrageenan-induced leg edema. Evaluation of antiarthritic activityof the ML terpenes rich fraction was carried out on collagen induced arthritis. Without finding any significant effect on back leg edema, corporal weight, arthritis histopathology severity scale or immunohistochemical tumoral necrosis factor alfa immunohistochemical evaluation (P>0.05). Discussion. Anti-inflammatory activity in TPAinduced acute ear edema model for CP, CA, and ML extracts and fractions can be interfered by phospholipase A2 related mediators due to similar effect with indomethacin was found. The ML terpens rich fraction neither show any anti-arthritic activity nor affected TNF-a expression on the autoimmune chronic arthritis model used, reason why it does not have immunomodulatory/anti-inflammatory effect on the evaluated dose. Conclusion. CA and CP terpenic rich fractions and ML ethanol extract showed significant anti-inflammatory activity in TPA-induced ear edema. They did have some activity in carrageenan-induced leg edema. The ML terpenic rich fraction did not have anti-arthritic activity in the collagen-induced arthritis model.

Caracteres anatômicos de folha e caule de Calea uniflora Less., Asteraceae / Anatomical characters of the leaf and stem of Calea uniflora Less., Asteraceae

Diversas espécies de Calea são utilizadas como medicinais e vários estudos químicos têm sido conduzidos para identificar compostos relevantes no gênero, no entanto, poucos enfocam aspectos morfoanatômicos. Este trabalho tem por objetivo caracterizar morfologicamente a folha e o caule de Calea uniflora Less., a fim de fornecer subsídios para a identificação da espécie. O material foi submetido às microtécnicas fotônica e eletrônica de varredura usuais. A lâmina foliar possui epiderme uniestratificada revestida por cutícula delgada e estriada. Em ambas as faces, ocorrem estômatos anomocíticos e anisocíticos, além de tricomas glandulares e tectores. Estes são pluricelulares e unisseriados, com ápice agudo. Os glandulares podem ser pluricelulares e plurisseriados ou capitados e inseridos em depressão na epiderme. O mesofilo é isobilateral e a nervura central é biconvexa, sendo percorrida por um feixe vascular colateral. O caule mostra secção circular e epiderme unisseriada, com tricomas similares aos da folha. Colênquima angular e clorênquima alternam-se no córtex, fibras perivasculares apõem-se ao floema, o arranjo vascular é colateral e a medula compõe-se de células parenquimáticas. Dutos secretores acompanham o sistema vascular na folha e no caule.

Various species of Calea are considered medicinal and several investigations have been carried out to identify relevant chemical substances in the genus, however, few studies have dealt with morpho-anatomical aspects. This work aimed to analyse the morphological characters of the leaf and stem of Calea uniflora Less., in order to contribute for the species identification. The plant material was prepared according to usual light and scanning microtechniques. The blade has uniseriate epidermis coated with thin and striate cuticle. Anomocytic and anisocytic stomata are seen on both surfaces, as well as glandular and non-glandular trichomes. The latter is pluricellular and uniseriate, with acute apex. The glandular trichomes are pluricellular and multiseriate or capitate and located in epidermal depression. The mesophyll is isobilateral, and the midrib is biconvex and traversed by a collateral vascular bundle. The stem shows circular transection and uniseriate epidermis, with trichomes similar to the leaf. Angular collenchyma and chlorenchyma alternate in the cortex, perivascular fibres adjoin the phloem, the vascular arrangement is collateral and the pith consists of parenchymatic cells. Secretory ducts are associated with the vascular system of the leaf and stem.