Anti-Acanthamoeba castellanii activity of alkaloid-enriched extracts and lycorine from the Amaryllidaceae species

Abstract Free-living amoebae of the genus Acanthamoeba are the causative agents of granulomatous encephalitis and keratitis, severe human infections. Bioactive compounds from plants are recognized as an alternative source for the development of new drugs. The Amaryllidaceae is a botanical family able to synthesize a very specific and consistent group of biologically active isoquinoline-like alkaloids. The alkaloidal fractions from the Brazilian species Hippeastrum canastrense, H. diniz-cruziae, H. puniceum, and Crinum x amabile, along with the alkaloid lycorine, were investigated against Acanthamoeba castellanii. The in vitro assays were performed with distinct concentrations of lycorine and alkaloidal fractions, while the cell viability was evaluated by the MTT method upon MDCK cells. Chlorhexidine 0.02% was used as the positive control. The effect of alkaloid fractions was concentration dependent, and 2000 µg mL-1 of H. canastrense and H. diniz-cruziae provided a 100% inhibition. At concentrations of 250, 500, and 1000 µg mL-1, the H. diniz-cruziae alkaloidal fraction showed the lowest cytotoxic effect (5%-7%) and remarkable anti-amoebic activity, demonstrating values of IC50 285.61 µg mL-1, low cytotoxicity (5%-7%), and selectivity index (7.0). Taken together, the results are indicative of the great potential that the alkaloids from H. diniz-cruziae have as new candidates for anti-amoebicidal compounds

Actividad cicatrizante de seis extractos hidroalcohólicos de plantas en heridas incisas de Rattus norvegicus albinus / Healing activity of six hydroalcoholic extracts of plants in incised wounds of Rattus norvegicus albinus

Introducción: En la medicina militar, los agentes cicatrizantes naturales frente a heridas por incisión son relevantes en el tratamiento de los militares. Objetivo: Evaluar la actividad cicatrizante de los extractos hidroalcohólicos de Peperomia congona Sodiro (congona), Annona muricata L. (guanábana), Urtica urens L. (ortiga),Ormosia coccinea (Aubl) Jacks (huayruro), Opuntia ficus-indica L. Mill. (tuna) y Musa acuminata Colla (plátano bellaco) en heridas incisas de Rattus norvegicus albinus (rata albina). Métodos: Estudio analítico experimental de tipo prospectivo y longitudinal. Se realizó el análisis fitoquímico preliminar de los 6 extractos. Se emplearon 80 ratas albinas machos, aleatorizadas y distribuidas en 8 grupos (n = 10). Se realizó una aplicación dermal durante 10 días consecutivos posteriores a la generación de heridas incisas que se realizaron en el dorso. Grupo I (cloruro de sodio al 0,9 por ciento). Grupo II (gel cicatrizante comercial). Del Grupo III al VIII fueron tratados con extracto hidroalcohólico de congona, guanábana, ortiga, huairuro, tuna y plátano bellaco respectivamente. La evolución de la cicatrización fue seguida en los días 1, 5 y 11 y medida en milímetros. Resultados: Se detectó la presencia de alcaloides, compuestos fenólicos y flavonoides en los 6 extractos; además de cumarinas en congona, taninos en guanábana, ortiga, tuna y huayruro, y quinonas en guanábana y plátano bellaco. Se comprobó la actividad cicatrizante en los extractos de guanábana (69,77 por ciento), tuna (66,27 por ciento), plátano bellaco (64,38 por ciento), ortiga (56,73 por ciento), congona (55,74 por ciento) y huayruro (54,50 por ciento), comparados con un gel comercial (72,21 por ciento). Conclusiones: El extracto hidroalcohólico de las hojas de Annona muricata L. (guanábana) presentó la mayor actividad cicatrizante en heridas incisas de ratas albinas machos(AU)

Introduction: In military medicine, natural healing agents against incision wounds are relevant in the treatment of the military. Objective: To evaluate the healing activity of the hydroalcoholic extracts of Peperomia congona Sodiro (congona), Annona muricata L. (guanabana), Urtica urens L. (ortiga), Ormosia coccinea (Aubl) Jacks (huairuro), Opuntia ficus-indica L. Mill. (tuna) and Musa acuminata Colla (platano bellaco) in incised wounds of Rattus norvegicus albinus (albino rat). Methods: Experimental analytical study of prospective and longitudinal type. The preliminary phytochemical analysis of the 6 extracts was carried out. We used 80 male albino rats, randomized and distributed in 8 groups (n = 10). A dermal application was made during 10 consecutive days after the generation of incised wounds that were made on the back. Group I (0.9 percent sodium chloride). Group II (commercial healing gel). From Group III to VIII they were treated with hydroalcoholic extract of congona, guanábana, ortiga, huairuro, tuna and platano bellaco respectively. The evolution of healing was followed on days 1, 5 and 11 and measured in millimeters. Results: The presence of alkaloids, phenolic compounds and flavonoids was detected in the 6 extracts; in addition to coumarins in congona, tannins in guanábana, nettle, tuna and huayruro, quinonas in guanábana and plantain bellaco. The healing activity was verified in the extracts of guanabana (69.77 percent), tuna (66.27 percent), platano bellaco (64.38 percent), ortiga (56.73 percent), congona (55.74 percent) and huayruro (54.50 percent), compared with a commercial gel (72.21 percent). Conclusions: The hydroalcoholic extract of the leaves of Annona muricata L. (guanábana) showed the highest healing activity in incised wounds of male albino rats(AU)

Punarnavine, an alkaloid isolated from ethanolic extract of Boerhaavia diffusa Linn. reverses depression-like behaviour in mice subjected to chronic unpredictable mild stress.

Punarnavine (20 and 40 mg/kg) and fluoxetine (20 mg/kg) per se administered orally for 14 successive days significantly decreased immobility periods of both unstressed and stressed mice in forced swim test. These drugs also significantly decreased sucrose preference in both stressed and unstressed mice as compared to their respective controls, indicating significant antidepressant-like activity. The drugs did not show any significant effect on locomotor activity of mice. The alkaloid also significantly decreased monoamine oxidase (MAO-A) activity, malondialdehyde levels in both unstressed and stressed mice; and significantly reversed the stress-induced decrease in reduced glutathione and catalase activity. It also significantly attenuated the stress-induced increase in plasma nitrite and corticosterone levels. Thus, punarnavine showed antidepressant-like activity in unstressed and stressed mice probably through inhibition of brain MAO-A activity, decrease in plasma nitrite levels and due to its antioxidant activity. In addition, punarnavine also showed antidepressant-like activity in stressed mice possibly through decrease in plasma corticosterone levels.

Tissue distribution & elimination of capsaicin, piperine & curcumin following oral intake in rats.

Background & objectives: Curcumin, capsaicin and piperine - the bioactive compounds present in spices-turmeric (Curcuma longa), red pepper (Capsicum annuum) and black pepper (Piper nigrum) respectively, have a considerable portion of structural homology. Tissue distribution and elimination of these three structurally similar bioactive compounds was examined following their oral intake in rats. Methods: Separate sets of animals (150 - 160 g) were orally administered the three spice principles at dosages of 30 mg (capsaicin), 170 mg (piperine) and 500 mg (curcumin) / kg body weight. The tissue concentrations of administered spice compounds were determined by HPLC. Results: Maximum distribution of 24.4 per cent of administered capsaicin was seen at 1 h, while no intact capsaicin was detectable after 4 days. Absorption of capsaicin was about 94 per cent and very rapid relative to other two compounds. A maximum of 10.8 per cent of administered piperine was seen in tissues at 6 h. Absorption of the administered piperine was about 96 per cent. Curcumin concentration was maximum in the intestine at 1 h; maximum in blood at 6 h and remained at significantly higher level even at 24 h. About 63.5 per cent of the curcumin dose was absorbed. Only a small portion of the administered dose of capsaicin (< 0.1%) and curcumin (0.173 %) was excreted in urine, whereas piperine was not detectable in urine. Enhanced bioavailability of curcumin was evidenced when the same was orally administered concomitant with piperine. Intestinal absorption of curcumin was relatively higher when administered concomitantly with piperine, and it stayed significantly longer in the body tissues. Intact curcumin was detected in brain at 24, 48 and 96 h with a maximum at 48 h. Conclusions: Considerable difference exists in the bioavailability of the three test compounds. Curcumin could be traced in the brain following its administration. Bioavailability of curcumin can be improved by co-administration with piperine.

Therapeutic effects of Sophora moorcroftiana alkaloids in combination with albendazole in mice experimentally infected with protoscolices of Echinococcus granulosus

The objective of the present study was to determine if the combination of alkaloids from Sophora moorcroftiana seeds and albendazole might be effective in the treatment of experimental echinococcosisin female NIH mice (6 weeks old and weighing 18-20 g, N = 8 in each group) infected withprotoscolices of Echinococcus granulosus. Viable protoscolices (N = 6 x 103) were cultured in vitro in 1640 medium and mortality was calculated daily. To determine the in vivo efficacy, mice were inoculated intraperitoneally with viable protoscolices and then treated once daily by gavage for three months with the alkaloids (50 mg kg-1 day-1) and albendazole (50 mg kg-1 day-1), separately and in combination (both alkaloids at 25 mg kg-1 day-1 and albendazole at 25 mg kg-1 day-1). Next, the hydatid cysts collected from the peritoneal cavity of the animals were weighed and serum IL-4, IL-2, and IgE levels were analyzed. Administration of alkaloids to cultured protoscolices showed significant dose- and time-dependent killing effects. The weight of hydatid cysts was significantly decreased upon treatment with each drug (P < 0.01), but the decrease was more prominent and the rate of hydatid cyst growth inhibition was much higher (76.1 percent) in the group receiving the combined treatments (18.3 ± 4.6 mg). IL-4 and total IgE were decreased (939 ± 447 pg/mL and 2.03 ± 0.42 IU/mL, respectively) in serum from mice treated with alkaloids and albendazole compared with the untreated control (1481 ± 619 pg/mL and 3.31 ± 0.37 IU/mL; P < 0.01). These results indicate that S. moorcroftiana alkaloids have protoscolicidal effects and the combination of alkaloids and albendazole has significant additive effects.

Efecto de los glicoalcaloides totales parcialmente purificados de flor de papa sobre las lesiones gástricas producidas por etanol en ratas albinas

El presente trabajo se realizó con la finalidad de determinar la diferencia del grado de lesiones gástricas entre el grupo de animales tratados con TGA y el grupo que no recibe tratamiento. Se llevó a cabo un estudio experimental, ciego, randomizado, comparativo y controlado por placebo en el Laboratorio de Fisiología de la Facultad de Medicina de la UNSA; utilizando ratas de la especie Novergicus, de la cepa Holtzman, de ambos sexos, sanos, con pesos de 178 gr. a 289 gr. en ayunas de 12h(grupo D) y de 16h(grupos A-B-C), mantenidas con agua "adlibitum" hasta dos horas antes del experimento. Las ratas fueron distribuídas aleatoriamente en cuatro grupos, con igual número de especímenes. El primer grupo(A) recibió sólo TGA(33mg/Kg); el grupo B(Control) recibió Etanol(5ml/Kg) y posteriormente agua; el grupo C(Tratamiento) recibió etanol + TGA y el grupo D(Prevención) recibió primero TGA y después Etanol. Todos los animales fueron sacrificados a las 6 horas de haber administrado el etanol, se les extrajo el estomago y se determinó el grado de daño de la mucosa gástrica con la ayuda de un microscópio estereoscópico a través de lectura ciega. Fueron guardados los estómagos en formol al 10 por ciento, siendo procesados por metodología convencional de H-E y luego, observados microscópicamente para determinar el daño de la mucosa. La comparación del grado de daño de la mucosa gástrica se realizó con las pruebas no paramétricas de Kruskall Wallis y Mann Whitney con ajuste de Bonferroni; encontrando diferencia significativa entre el grupo control(B) y el grupo tratamiento (C); siendo iguales el grupo control con el grupo de prevención (D). Se concluye que los TGA de la papa a dosis de 33mg/Kg tienen efecto terapeútico en las lesiones gástricas producidas por etanol en ratas albinas

Toxicity of pure alkaloid of Tylophora asthamatica in male rat.

Administration of pure alkaloid of T. asthamatica, suspended in peanut oil and given in single doses (12-100 mg/kg) by gavage, to male rats caused inactivity, respiratory distress, salivation, nasal discharge and diarrhoea. The oral LD50 value of the alkaloid was 35.32 mg/kg. In short term toxicity study daily doses of the alkaloid (1.25, 2.5, 5 and 10 mg/kg) were given to male rats for 15 days. Smaller doses of the alkaloid (1.25 and 2.5 mg/kg/day) produced no signs of poisoning or death in animals; while 5 mg/kg/day produced signs of poisoning and death of two animals, 10 mg/kg/day caused death of all the animals within 7 days. Activities of glutamic oxaloacetic transaminase, glutamic pyruvic transaminase and alkaline phosphatase were significant and associated with morphological changes in liver. The alkaloid also caused marked changes in the morphology of seminiferous tubules and spermatogenic activity of experimental animals. Since the alkaloid is effective in microgram quantities, the non toxic effects observed after daily doses of 1.25 mg/kg in male rats assume great therapeutic significance.