Volatile Components, Antioxidant and Phytotoxic Activity of the Essential Oil of Piper acutifolium Ruiz & Pav. from Peru.

Piper acutifolium Ruiz & Pav is known as "matico" and belongs to the Piperaceae family, and in Peru it is traditionally used as an infusion or decoction to ameliorate wound healings or ulcers. In this study, the aim was to investigate the volatile components, the antioxidant profile, and the phytotoxic activity of the essential oil (EO) of P. acutifolium from Peru. To identify the phytoconstituents, the EO was injected into a Gas Chromatography-Mass Spectrometry (GC-MS) to obtain the chemical profile of the volatile components, followed by the antioxidant activity carried out by the reaction with three organic radicals (2,2-diphenyl-1-picrylhydrazyl (DPPH); 2,2'-azinobis-(3-ethylbenzothiazoline)-6- sulfonic acid (ABTS); ferric reducing/antioxidant power (FRAP)). Finally, the phytotoxic capabilities of the EO were tested on two model plants, Lactuca sativa seeds and Allium cepa bulbs. As a result, the analysis identified α-phellandrene as its main volatile chemical at 38.18%, followed by ß-myrcene (29.48%) and ß-phellandrene (21.88%). Regarding the antioxidant profile, the half inhibitory concentration (IC50) in DPPH was 160.12 ± 0.30 µg/mL, for ABTS it was 138.10 ± 0.06 µg/mL and finally in FRAP it was 450.10 ± 0.05 µg/mL. The phytotoxic activity demonstrated that the EO had high activity at 5% and 10% against L. sativa seed germination, the inhibition of root length, and hypocotyl length. Additionally, in A. cepa bulbs, the inhibition root length was obtained at 10%, both comparable to glyphosate, which was used as a positive control. The molecular docking on 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) revealed that α-phellandrene had -5.8 kcal/mol, being near to glyphosate at -6.3 kcal/mol. The conclusion shows that the EO of P. acutifolium presented antioxidant and phytotoxic activity and might be useful as a bioherbicide in the future.

Eficacia clínica del extracto de Matico (Piper angustifolium) en la evolución de la cicatrización de heridas por estomatitis aftosa recidivante / Clinical efficacy of Matico extract (Piper angustifolium) in the evolution of wound healing due to recurrent aphthous stomatitis

La estomatitis aftosa recurrente (EAR) es un trastorno ulcerativo doloroso cíclico que comúnmente afecta la mucosa oral. Factores locales y sistémicos como trauma, sensibilidad alimentaria, deficiencias nutricionales, condiciones sistémicas y desórdenes inmunológicos están asociados con el desarrollo de la enfermedad. Debido a las similitudes en las características de las úlceras pépticas y las úlceras aftosas orales, parece razonable suponer que Helicobacter pylori podría desempeñar un papel en el desarrollo de la EAR. Las especies de Piper son plantas aromáticas que se utilizan como especias en la cocina, pero sus metabolitos secundarios también han mostrado efectos biológicos sobre la salud humana. El objetivo del presente estudio fue determinar la relación entre H. pylori y EAR y el efecto cicatrizante del extracto Matico sobre las ulceras bucales. Se reclutó a cuarenta sujetos con EAR. Los participantes fueron asignados al azar al grupo de prueba o al grupo de control. El tamaño y número de las úlceras se registraron el día 1 (valor inicial) y el día 30 (valor final). El tamaño de la ulceras, presencia de pseudomembranas y halos eritematosos fueron estadisticamente significativas en relación a la positividad de H. pylori (p <0,05). No hubo diferencias significativas entre la morfología, número y localización de las aftas (p> 0,05). El grupo que recibió el extracto tuvo un porcentaje de mejoría en la reducción del número de aftas del 65% el día final de evaluación en comparación de sólo un 25% del grupo que no recibió el extracto Matico(AU)

Recurrent aphthous stomatitis (RAS) is a cyclic painful ulcerative disorder that commonly affects the oral mucosa. Local and systemic factors such as trauma, food sensitivity, nutritional deficiencies, systemic conditions, immunological disorders and genetic polymorphisms are associated with the development of the disease. Because of similarities in the characteristics of peptic ulcers and oral aphthous ulcers, it seems reasonable to hypothesize that Helicobacter pylori could play a role in the development of RAS. Piper species are aromatic plants used as spices in the kitchen, but their secondary metabolites have also shown biological effects on human health. The aim of the present study was to determine the relationship between H. pylori and RAS and the healing effect of Matico extract on mouth ulcers. Fuorty subjects with RAS were recruited. The participants were randomly allocated to the test group or the control group. The size and number of ulcers were recorded on day 1 (initial value) and day 30 (final value). The size of the ulcers, the presence of pseudomembranes and erythematous halos were statistically significant in relation to the positivity of H. pylori (p <0.05). There were no significant differences between the morphology, number and location of the canker sores (p> 0.05). The group that received the extract had a percentage improvement in the reduction of the number of canker sores of 65% on the final day of evaluation compared to only 25% of the group that did not receive the Matico extract(AU)

Polyphenolic Compounds Extracted and Purified from Buddleja Globosa Hope (Buddlejaceae) Leaves Using Natural Deep Eutectic Solvents and Centrifugal Partition Chromatography.

Chemical profiling of Buddleja globosa was performed by high-performance liquid chromatography coupled to electrospray ionization (HPLC-DAD-ESI-IT/MS) and quadrupole time-of-flight high-resolution mass spectrometry (HPLC-ESI-QTOF/MS). The identification of 17 main phenolic compounds in B. globosa leaf extracts was achieved. Along with caffeoyl glucoside isomers, caffeoylshikimic acid and several verbascoside derivatives (ß-hydroxyverbascoside and ß-hydroxyisoverbascoside) were identified. Among flavonoid compounds, the presence of 6-hydroxyluteolin-7-O-glucoside, quercetin-3-O-glucoside, luteolin 7-O-glucoside, apigenin 7-O-glucoside was confirmed. Campneoside I, forsythoside B, lipedoside A and forsythoside A were identified along with verbascoside, isoverbascoside, eukovoside and martynoside. The isolation of two bioactive phenolic compounds verbascoside and forsythoside B from Buddleja globosa (Buddlejaceae) was successfully achieved by centrifugal partition chromatography (CPC). Both compounds were obtained in one-step using optimized CPC methodology with the two-phase solvent system comprising ethyl acetate-n-butanol-ethanol-water (0.25:0.75:0.1:1, v/v). Additionally, eight Natural Deep Eutectic Solvents (NADESs) were tested for the extraction of polyphenols and compared with 80% methanol. The contents of verbascoside and luteolin 7-O-glucoside after extraction with 80% methanol were 26.165 and 3.206 mg/g, respectively. Among the NADESs tested in this study, proline- citric acid (1:1) and choline chloride-1, 2- propanediol (1:2) were the most promising solvents. With these NADES, extraction yields for verbascoside and luteolin 7-O-glucoside were 51.045 and 4.387 mg/g, respectively. Taken together, the results of this study confirm that CPC enabled the fast isolation of bioactive polyphenols from B. globosa. NADESs displayed higher extraction efficiency of phenolic and therefore could be used as an ecofriendly alternative to classic organic solvents.

Buddleja globosa (matico) prevents collagen-induced platelet activation by decreasing phospholipase C-gamma 2 and protein kinase C phosphorylation signaling.

Platelets play a key role in thrombosis and cardiovascular diseases. Medicinal plants could be one of the most important factors that influence risks for platelet activation. Buddleja globosa (known as "matico") is a medicinal plant with many biological activities. The high content of polyphenols suggest that matico could have antiplatelet activity. The present study was aimed at evaluating mechanisms of antiplatelet action of an extract of matico. We demonstrated that matico extract at low concentrations and in a concentration dependent manner (0.05-1 mg/mL) was a potent inhibitor of platelet aggregation in response to collagen, convulsion and ADP (IC50 values was 61 µg/mL, 72 µg/mL and 290 µg/mL, respectively). In this sense matico extract exerted the greatest antiaggregant activity induced by collagen. Similarly, matico showed a decrease in % of positive platelet for P-selectina (vehicle, 0.01, 0.05, 0.1, 0.5 and 1 mg/mL were 32 ± 2%, 29 ± 2 (p < 0.05), 19 ± 1 (p < 0.01), 15 ± 2 (p < 0.01), 10 ± 1% (p < 0.01) and 7 ± 2% (p < 0.01), respectively) and PAC-1 binding (vehicle, 0.01, 0.05, 0.1, 0.5 and 1 mg/mL were 59 ± 1, 58 ± 3 (n.s), 55 ± 2 (p < 0.05), 50 ± 2 (p < 0.01), 38 ± 1 (p < 0.01), 36 ± 2 (p < 0.01). The cellular mechanism for the antiplatelet activity of matico might be mediated by the inhibition of phospholipase C-gamma 2 and protein kinase C phosphorylation. This beneficial property of matico may be of importance in thrombosis, in which platelet activation and aggregation are important determinants of thrombus initiation and development, and may contribute to the beneficial effects of matico intake in the prevention of cardiovascular diseases.

Efecto del extracto acuoso de Piper elongatum Vahl. (matico) sobre la motilidad intestinal en ratones / Effect of aqueous extract from Piper elongatum Vahl (matico) on the intestinal motility in mice

Objetivo: evaluar el efecto del extracto acuoso de Piper elongatum Vahl. (matico) sobre la motilidad intestinal en ratones BALB/c. Métodos: estudio experimental. Se empleó el método del carbón activado. Los grupos fueron: Grupo 1 control negativo con NaCl 9.0 por ciento, Grupo 2 control positivo con loperamida 10 mg/Kg, grupos tratamiento de la planta a dosis de 125 mg/Kg (Grupo3), 250 mg/Kg (Grupo4) y 500 mg/Kg (Grupo5). Se utilizaron las pruebas estadísticas de ShapiroWilk, ANOVA y Bonferroni; se consideró p<0,05 como significativo e intervalo de confianza 95 por ciento. Resultados: el porcentaje del intestino recorrido por el carbón activado fue de 69,4±4,5; 24,9±3,0; 75,3±4,3; 71,1±4,5; 56,4±2,7 y 59,4±3,0 en los grupos del 1 al 5, respectivamente. La prueba post hoc Bonferroni no mostró diferencias entre el control negativo y los grupos tratamiento con Piper elongatum Vahl., mientras que sí existió diferencia entre el control positivo con los demás grupos y entre el grupo 3 y 5 (p<0,001). Conclusiones: el extracto acuoso de las hojas de Piper elongatum Vahl. no posee efecto sobre la motilidad intestinal a las dosis evaluadas, se requieren más estudios(AU)

Objective: to evaluate the effect of aqueous extract from Piper elongatum Vahl (matico) on the intestinal motility in BALB/c mice. Methods: experimental study that used activated carbon. The involved groups were: Group 1 with negative control with 9.0 percent NaCl; Group 2 positive control with loperamide 10mg/Kg, groups treated with the plant at doses of 125 mg/kg (Group3), 250 mg/kg (Group4) and 500 mg/Kg (Group5). ShapiroWilk, ANOVA and Bonferroni statistical tests were used; p<0.05 was considered as significant and 95 percent confidence interval. Results: the percentages of intestine explored with the active charcoal were 69,4±4,5; 24,9±3,0; 75,3±4,3; 71,1±4,5; 56,4±2,7 and 59,4±3,0 in groups 1 to 5 respectively. The ANOVA on this variable showed a p<0,001. The post hoc Bonferroni test showed no differences between the negative control and treatment groups with Piper elongatum Vahl whereas there was a difference between the positive control and the other groups and between the groups 3 and 5 (p<0.001). Conclusions: the aqueous extract from Piper elongatum Vahl has no effect on the intestinal motility at the evaluated doses, so further studies are needed(AU)

Protective Effect of Piper aduncum Capsule on DMBA-induced Breast Cancer in Rats.

The possible protective effect of Piper aduncum capsule on DMBA (dimethylbenz[α]anthracene)-induced breast cancer in rats was assessed by monitoring the tumor and lung metastases incidence and recording hematological and biochemical parameters and frequency of micronuclei. Mammary carcinogenesis was induced in 36 female Holtzman rats by providing a single subcutaneous injection of DMBA. Oral administration of P. aduncum capsule lowered adenocarcinoma and lymph node metastases incidence. Pulmonary metastasis was significantly lowered (P < 0.05). Hematological indicators showed that the triglyceride level was significantly lowered (P < 0.01) and high-density lipoprotein (HDL) level was significantly increased (P < 0.01). Also, P. aduncum capsule significantly lowered the C reactive protein (CRP) level (P < 0.01) and malondialdehyde level (P < 0.05). There was a significant decrease in the frequency of DMBA-induced micronucleated polychromatic erythrocyte (P < 0.01). Considering the antitumorigenic, hypolipidemic, anti-inflammatory, antioxidant, and antigenotoxic properties of P. aduncum capsule, we conclude that it has a protective effect on DMBA-induced breast cancer in rats.

Efecto gastroprotector y antisecretor de un fitofármaco de hojas de matico (Piper aduncum) / Gastroprotective and antisecretory effect of a phytochemical made from matico leaves (Piper aduncum)

Objetivos. Determinar el efecto gastroprotector y antisecretor del extracto etanólico de las hojas de matico (Piper aduncum) en modelos animales. Materiales y métodos. Para la evaluación del efecto gastroprotector se utilizó 220 ratones de la cepa Balb C57, los cuales fueron aleatorizados en 22 grupos de diez animales, a los cuales se les indujo la formación de úlceras gástricas con indometacina, la gastroprotección se determinó a través de tres aspectos: inflamación, número de bandas hemorrágicas y número de úlceras. Para evaluar el efecto antisecretor se utilizó 64 ratas albinas machos Holtzman, los cuales fueron aleatorizados en ocho grupos de ocho animales, un control y siete grupos de tratamiento con un nivel de dosis de los extractos y dos niveles de dosis en los fitofármacos; la antisecreción se realizó con el ensayo de ligazón pilórica. Resultados. Para la gastroprotección, los extractos de diclorometano, cloroformo, hexano y metanol, lograron una disminución de la inflamación de más del 66% (p<0,05); el extracto etanólico presenta una actividad de 100% para disminuir el número de bandas hemorrágicas (p<0,05); el extracto clorofórmico presenta una actividad antiulcerosa de 75% (p<0,05). Respecto a la antisecreción, el fitofármaco en cápsulas conteniendo el extracto etanólico logró un 72% de reducción del volumen de la secreción gástrica (p<0,01) y un incremento del pH en 104,3% (p<0,01). Conclusiones. En condiciones experimentales los extractos etanólico, sus fracciones y su fitofármaco son gastroprotectores en ratones y antisecretores en ratas.

Objectives. To determine the gastroprotective and antisecretory effect of ethanol extract from matico leaves (Piper aduncum) in animal models. Materials and methods. To evaluate the gastroprotective effect, 220 mice of the Balb C57 strain were used. They were randomized in 22 groups of ten animals each, in which the formation of gastric ulcers was induced with indomethacin. Gastroprotection was determined by evaluating three aspects: inflammation, number of hemorrhagic shocks and number of ulcers. To evaluate the antisecretory effect, 64 white male Holtzman rats were used, which were randomized in eight groups of eight animals, one control and seven groups of treatment with one extract dose level and two phytochemical dose levels. Antisecretion was obtained through the pylorus ligation. Results. Regarding gastroprotection, dichloromethane, chloroform, hexane and methanol extracts decreased inflammation to over 66% (p<0,05). The ethanolic extract shows 100% activity in reducing the number of hemorrhagic bands (p<0,05). The chloroform extract shows antiulcer activity at 75% (p<0,05). In terms of antisecretion, the phytochemical in capsules containing the ethanolic extract achieved 72% reduction of the gastric secretion volume (p<0,01) and 104,3% (p<0,01) PH increase. Conclusions. In experimental conditions, ethanolic extracts, their fractions and phytochemicals have a gastroprotective effect in mice and antisecretory effect in rats.

Antifungal activity of raw extract and flavanons isolated from Piper ecuadorense from Ecuador

The MeOH extract of Piper ecuadorense Sodiro, Piperaceae, was chosen for metabolite isolation and elucidation due to the strong antifungal activity exhibited, measured by means of the broth microdilution method. Two known flavonoids: pinostrobin (1) and pinocembrin (2) were isolated from 4.16 g. of dichloromethane extract by column chromatography, using a gradient of hexane/EtOAc. A total of 20 mg of 1 were obtained from the fraction eluted with hexane-EtOAc 95:5 v/v, and 100 mg of 2 were obtained from the fraction eluted with hexane-EtOAc 85:15 v/v. The MIC values of the MeOH extract was 31.25 µg/mL for Trichophyton mentagrophytes ATCC® 28185 and 62.5 µg/mL for Trichophyton rubrum ATCC® 28188. The MIC value of pinocembrin was 125 µg/mL for Trichophyton mentagrophytes ATCC® 28185 and Trichophyton rubrum ATCC® 28188. Pinostrobin in antifungal test was not active against fungi tested.

Efecto antihipertensivo del extracto de Piper aduncum ‘matico’ sobre la hipertensión inducida por L-NAME en ratones / Piper aduncum ‘matico’ extract antihypertensive effect on L-NAME induced hypertension in mice

Introducción: El Piper aduncum es una planta conocida como matico. Se le atribuye efectos antihipertensivo, antiinflamatorio, cicatrizante. Objetivos: Determinar el efecto antihipertensivo del extracto de Piper aduncum ‘matico’, sobre la hipertensión inducida por L-NAME, en ratones. Diseño: Experimental. Lugar: Facultades de Medicina y de Farmacia y Bioquímica, Universidad Nacional Mayor de San Marcos, Lima, Perú. Material biológico: Extracto etanólico de las hojas de Piper aduncum y ratas. Intervenciones: Se utilizó seis grupos de seis ratones Muss musculus cada uno, uno sin hipertensión (control negativo) y cuatro con hipertensión inducida por L-NAME: un control positivo y tres grupos para las dosis de 50, 150 y 300 mg/kg, respectivamente. El tratamiento se realizó por vía oral, una vez por día, durante 25 días. Las mediciones de la presión arterial sistólica (PAS), presión arterial diastólica (PAD) y presión arterial media (PAM) fueron realizadas dos veces por semana (martes y viernes) y se consideró las mediciones entre los días 19 y 23 de iniciado el tratamiento. Principales medidas de resultados: Actividad antihipertensiva. Resultados: En los días 19 y 23 se observó los mejores niveles de presión arterial en el grupo control y los experimentales, correspondiendo a las mediciones 6 y 7. La eficacia antihipertensiva para enalapril fue 24,1 a 20,6%, respectivamente, seguida por matico, entre 24,9 y 13,7% (p<0,05). Conclusiones: En las condiciones experimentales, se demostró la actividad antihipertensiva del extracto etanólico de hojas de Piper aduncum ‘matico’.

Introduction: Piper aduncum also known as matico is credited with antihypertensive, anti-inflammatory, healing effects. Objectives: To determine the antihypertensive activity of Piper aduncum ‘matico’ extract on L-NAME-induced hypertension in mice. Design: Experimental. Setting: Faculty of Medicine, and Faculty of Pharmacy and Biochemistry, National University of San Marcos, Lima, Peru. Biological material: Piper aduncum ethanolic extract leaves and mice. Interventions: Six groups of six Muss musculus mice each were used, one without hypertension (negative control) and four with L-NAME-induced hypertension: one positive control and three groups for 50, 150 and 300 mg/kg doses, respectively. Treatment was given orally once daily for 25 days. Measurements of systolic blood pressure (SBP), diastolic blood pressure (DBP) and mean arterial pressure (MAP) were performed twice weekly (Tuesday and Friday) and measurements between 19 and 23 days of treatment were considered. Main outcome measures: Antihypertensive activity. Results: Higher blood pressure levels were observed at days 19 and 23 in the control and experimental groups that corresponded to measurements 6 and 7. The antihypertensive efficacy to enalapril was 24.1 to 20.6%, followed by matico between 24.9 and 13.7% (p <0.05). Conclusions: Antihypertensive activity of Piper aduncum ‘matico’ extract was demonstrated under experimental conditions.