Oxytocin neuron activation by acute infusion of Montanoa genus plants in the Wistar rats / Activación de neuronas de oxitocina por la infusión aguda de plantas del género de Montanoa en la rata Wistar

In tradition al Mexican medicine, plants from the Montanoa genus, family Asteraceae ( Montanoa tomentosa , Montanoa grandiflora , and Montanoa frutescens ) have been used to induce labor owing to their uterotonic properties like those produced by oxytocin (OXT). However, w hether infusions of these plants can activate hypothalamic OXT - producing neurons is unknown. To test this possibility, five independent groups of Wistar rats (n=4) were included: intact, vehicle, and three groups that received 50 mg/kg p.o. of M. tomentosa , M. grandiflora , and M. frutescens infusions, respectively. Ninety min after treatment, the brains were obtained and processed using double - labeled immunohistochemistry for Fos protein and oxytocin (Fos/OXT - ir). Rats that received Montanoa infusions had s ignificantly greater number of Fos/OXT - ir cells in the paraventricular (PVN) and supraoptic (SON) nuclei, with respect to intact and vehicle groups. These findings demonstrate that Montanoa infusions activated OXT neurons, an effect that may be related to the reported pharmacological properties.

En la medicina tradicional mexicana, plantas del género Montanoa , familia Asteraceae ( Montanoa tomentosa , Montanoa grandiflora y Montanoa frutescens ), se han utilizado para inducir el parto debido a sus propiedades uterotónicas, aparentemente similares a las producidas por la hormona oxitocina (OXT). Sin embargo, se desconoce si las infusiones de estas plantas pueden activar neuronas hipotalámicas productoras de OXT. Para probar esta posibilidad, se incluyeron cinco grupos independientes (n=4): intacto, vehículo y tres grupos que recibieron 50 mg/kg p.o. de infusiones de M. tomentosa , M. grandiflora , y M. frute scens , respectivamente. Noventa minutos después del tratamiento, los cerebros fueron obtenidos y procesados por doble marcaje de inmunohistoquímica para la proteína Fos y oxitocina (Fos/OXT - ir). Las ratas que recibieron infusiones de Montanoa aumentaron si gnificativamente el número de células Fos/OXT - ir en los núcleos paraventricular (PVN) y supraóptico (SON), respecto a los grupos intacto y vehículo. Estos hallazgos demuestran que las infusiones de Montanoa activan neuronas de OXT, lo que podría estar rela cionado con sus propiedades farmacológicas

Antibacterial Activity and Reversal of Multidrug Resistance of Tumor Cells by Essential Oils from Fresh Leaves, Flowers, and Stems of Montanoa quadrangularis Schultz Bipontinus (Asteraceae) Collected in Mérida-Venezuela.

Essential oils obtained by hydrodistillation of Montanoa quadrangularis leaves, flowers, and stems were analyzed by GC and GC/MS techniques revealing myrcene, limonene, ß-phellandrene, and sabinene among the main components. The aim of the present study was to evaluate the MDR modulator activity on human MDR1 gene transfected mouse lymphoma cell line and the antimicrobial activity on the essential oils obtained from different parts of the species under investigation. The results revealed that MQL caused a similar increase in the fluorescence activity of the cells at 0.02 µL/mL comparing to the Verapamil® value. The antimicrobial assay was carried out according to the disc diffusion method. Five different bacterial strains (Staphylococcus epidermidis, Bacillus subtilis, Pseudomonas aeruginosa, Escherichia coli AG 100, and Escherichia coli AG100A) were treated with the essential oils and the zones of inhibition were determined on TSA plates and TSA agar plates supplemented with Tween 20. MQF and MQL showed activity against B. subtilis, S. epidermidis, and E. coli AG 100A while MQS was only active against E. coli AG 100A on TSA agar plates experiment. In case of TSA agar plates supplemented with 0.1 v/v% Tween 20 detergent, MQF showed inhibition on B. subtilis, S. epidermidis, and E. coli AG 100A; MQL was active against B. subtilis, E. coli AG 100, and E. coli AG 100A while MQS was only active against E. coli AG 100A.

Distribución potencial de dos subespecies de Montanoa tomentosa (Asteracerae) en México / Potential distribution of two subspecies of Montanoa tomentosa (Asteracerae) in Mexico

Introducción: la Montanoa tomentosa es un recurso fitogenético con propiedades medicinales utilizado en México. A pesar de su relevancia botánica, la información específica de las distribuciones de taxones infraespecíficos y la los factores que influyen en su desarrollo aún son limitados. Objetivo: Informar el potencial geográfico distribución y las variables ambientales asociadas a nichos ecológicos de dos subespecies de M. tomentosa (M. tomentosa subsp. Tomentosa y M. tomentosa subsp. Xanthiifolia) en territorio mexicano. Métodos: A Se creó una base de datos de registros de herbario, se realizó una búsqueda sistemática in situ de ambas subespecies desde Octubre de 2015 a marzo de 2016, durante la temporada de floración y fructificación. Posteriormente, recorridos exploratorios (Guanajuato, Querétaro, Hidalgo, Ciudad de México, Estado de México, Puebla, Oaxaca y Chiapas). Un total de 181 Se muestrearon y georreferenciaron individuos, generando nuevos registros para la actualización de la base de datos. Tres hábitat modelos de idoneidad (complejo M. tomentosa, M. tomentosa subsp. tomentosa y M. tomentosa subsp. xanthiifolia) se generaron utilizando el software MaxEnt. Este software estima la probabilidad de distribución de especies. utilizando datos georreferenciados y características ambientales como predictores, mediante el logaritmo de entropía máxima y el método bayesiano. Resultados: De 22 variables ambientales utilizadas como predictores, cinco (estacionalidad de temperatura, isotermalidad, temperatura máxima del mes más cálido, precipitación del más cálido trimestre y precipitación del trimestre más húmedo) contribuyeron con el 70,2% de la contribución porcentual total. los El modelo mostró una distribución potencial de 77 688 km2, aproximadamente el 4% del territorio de la República Mexicana. Ambos La subespecie creció entre 500 y 2 500 m. Se asociaron a vegetación xerófila y heliófila, a veces también estaban ubicadas en la periferia o incrustadas en aberturas de bosques templados y subhúmedos. No estuvieron presentes regiones secas del norte del país. Conclusiones: estacionalidad de la temperatura, isotermalidad y la temperatura máxima del mes más cálido determinan la distribución de la especie. Precipitación del cuarto más cálido y la precipitación del cuarto más húmedo define el hábitat entre M. tomentosa subsp. tomentosa y M. tomentosa subsp. xanthiifolia. Estos hallazgos aclaran las preferencias ecológicas y la delimitación los taxones infraespecíficos de M. tomentosa.

Introduction: Montanoa tomentosa is a phytogenetic resource with medicinal properties used in Mexico. Despite its botanic relevance, the specific information of infraespecific taxa distributions and the environmental factors that influence their development is still limited. Objective: To report the potential geographic distribution and the environmental variables associated to ecological niches of two subspecies of M. tomentosa (M. tomentosa subsp. tomentosa and M. tomentosa subsp. xanthiifolia) in the Mexican territory. Methods: A database of herbarium records was created, an in situ systematic search of both subspecies was carried out from October 2015 to March 2016, during the flowering-fruiting season. After that, exploratory tours (Guanajuato, Querétaro, Hidalgo, Mexico City, State of Mexico, Puebla, Oaxaca and Chiapas) were realized. A total of 181 individuals were sampled and georeferenced, generating new records for the database upgrading. Three habitat suitability models (M. tomentosa complex, M. tomentosa subsp. tomentosa and M. tomentosa subsp. xanthiifolia) were generated using the MaxEnt software. These software estimates the probability of species distribution using geo-referenced data and environmental characteristics as predictors, through the maximum entropy logarithm and the Bayesian method. Results: From 22 environmental variables used as predictors, five (seasonality of temperature, isothermality, maximum temperature of the warmest month, precipitation of the warmest trimester and precipitation of the wettest trimester) contributed 70.2 % of the total percentage contribution. The model showed a potential distribution of 77 688 km2, approximately 4 % of the Mexican Republic territory. Both subspecies grew between 500 and 2 500 m. They were associated to xerophilous and heliophiles vegetation, sometimes they were located in the periphery or embed in openings of temperate and sub-humid forests too. They were not present dry regions from the North of the country. Conclusions: Temperature seasonality, isothermality and maximum temperature of the warmest month determine the distribution of the species. Precipitation of the warmest quarter and precipitation of wettest quarter defines the habitat between M. tomentosa subsp. tomentosa and M. tomentosa subsp. xanthiifolia. These findings clarify the ecological preferences and delimitation the infraspecific taxa of M. tomentosa.

GABAA/benzodiazepine receptor complex mediates the anxiolytic-like effect of Montanoa tomentosa.

ETHNOPHARMACOLOGICAL RELEVANCE: Montanoa tomentosa also named Cihuapatli is a native plant of Mexico that has been used in traditional medicine for the last five centuries mainly as a remedy for reproductive impairments. However, there are reports indicating that this plant was also consumed by Mexican ancient people for its relaxing properties. In order to corroborate this information, the present study was conducted to evaluate the effect of Montanoa tomentosa lyophilisate (MT) on rat׳s anxiety-like behavior and to analyze its mechanism of action. MATERIALS AND METHODS: The anxiolytic-like action of MT (1.5, 3.0, 6.0 and 12.0 mg/kg) was investigated in male Wistar rats tested in three animal models of anxiety: the burying behavior, the elevated plus maze and the hole-board tests. As a positive control, the anti-anxiety effects of different doses of the selective GABAA receptor agonist muscimol were also analyzed. In order to evaluate the participation of the GABAA and oxytocin receptors in the anxiolytic-like actions of MT, the GABAA receptors blockers picrotoxin (0.25 and 0.50 mg/kg), bicuculline (2.0 mg/kg) and flumazenil (5.00 and 10.0 mg/kg), the neurosteroid inhibitor finasteride (50.0 and 100 mg/kg) and the oxytocin receptor antagonist atosiban (0.25 µg) were used. Finally, to evaluate general activity, and motor coordination, the open field and rota-rod tests were used. RESULTS: MT at 3.0 mg/kg showed anxiolytic-like effects in the three anxiety paradigms without affecting reactivity, general motor activity or motor coordination; however, at higher doses sedative effects were observed. Picrotoxin (0.25 and 0.50 mg/kg), flumazenil (10.0 mg/kg) and finasteride (100 mg/kg) antagonized the anxiolytic-like actions of MT in the burying behavior test. In the plus maze and hole-board tests bicuculline (2.0 mg/kg) blocked the effects of the plant as well. Atosiban (0.25 µg) did not antagonize the anxiolytic-like actions of MT. CONCLUSIONS: The results corroborate the anxiolytic-like actions of Montanoa tomentosa and suggest that this effect is mediated through GABAA receptors but not oxytocin receptors.

Anti-fertility and other biological activities of zoapatle (Montanoa spp. )with biotechnological application / Anti-fertilidad y otras actividades biológicas del zoapatle (Montanoa spp. ) con aplicación biotecnológica

Species from the Montanoa genus have been used for hundreds of years in Mexican traditional medicine. The Náhuatl name given by Aztecs to this plant was “cihuapahtli”, meaning “woman’s medicine” (cihua, woman; pahtli, medicine), because M. tomentosa has been used to treat female health disorders since pre-Hispanic times due to its uterotonic properties. Later on, the Spanish pronunciation transformed the word into “zoapatle”, the name still used by herb sellers in México. Despite its use in folk medicine, intensive scientific research on this plant did not begin until the early 1970’s. The only two available reviews exclusively focused on the Montanoa species cover the period from 1529 to 1985. However, in the last two decades, important phytochemical, biochemical and biotechnological advances were achieved. This is a review of the history, botany, most relevant chemistry, biological activity and biotechnology of the secondary metabolites from the Montanoa genus so far.

Especies del género Montanoa han sido empleadas por siglos en la medicina tradicional Mexicana. El nombre asignado a esta planta por los Aztecas fue “cihuapahtli”, ó “medicina para la mujer” (cihua, mujer; pahtli, medicina). Específicamente, M. tomentosa es usada para tratar desordenes de la mujer desde tiempos prehispánicos por sus propiedades uterotónicas. La castellanización transformó esta palabra en “zoapatle”, este nombre sigue siendo usado por los curanderos en México. A pesar de su uso en la medicina folklórica, la investigación científica intensa en esta planta comenzó desde inicios de 1970. Las únicas dos revisiones disponibles y dedicadas exclusivamente a las especies de Montanoa cubren el periodo de 1529 a 1985. Sin embargo, en las últimas dos décadas, se han logrado importantes avances fitoquímicos, bioquímicos y biotecnológicos. Este trabajo presenta una revisión de la historia, botánica, química más relevante, actividades biológicas y biotecnología de los metabolitos secundarios del género Montanoa a la fecha.

Montanoa frutescens and Montanoa grandiflora extracts reduce anxiety-like behavior during the metestrus-diestrus phase of the ovarian cycle in Wistar rats.

In previous studies, the anxiolytic-like effects of Montanoa tomentosa and Montanoa frutescens were reported in male rats, but the potential anxiolytic-like effects of Montanoa plants during the different phases of the ovarian cycle in rats remain to be explored. The anxiolytic-like effects of the aqueous crude extracts of M. frutescens (25 and 50 mg/kg) and M. grandiflora (25 and 50 mg/kg) in the elevated plus maze were investigated in Wistar rats during the estrous cycle and compared with 2 mg/kg diazepam as a reference anxiolytic drug. To investigate any motor effect (i.e., hyperactivity, no changes, or hypoactivity) associated with the treatments, the rats were evaluated in the open field test. The M. frutescens (25 and 50 mg/kg) and M. grandiflora (50 mg/kg) extracts exerted anxiolytic-like effects during the metestrus-diestrus phase, similar to diazepam, without disrupting spontaneous motor activity. No significant effects of the extracts were detected in either behavioral test during the proestrus-estrus phase, whereas diazepam produced motor hypoactivity in the open field test. These results indicate that the M. frutescens and M. grandiflora extracts possess anxiolytic-like effects that depend on the ovarian cycle phase, supporting the Mexican ancient medicinal use of these plants to ameliorate anxiety disorders.

Montanoa tomentosa glandular trichomes containing kaurenoic acids chemical profile and distribution.

Montanoa tomentosa has been used in traditional medicine in Mexico to treat diverse female health disorders; it is particularly useful in inducing childbirth. Microscopic analysis of leaf surfaces of M. tomentosa revealed the presence of glandular trichomes. The chemical profile and distribution of glandular trichomes from different developmental stages of M. tomentosa leaves were investigated. Two diterpenic acids, kaurenoic and grandiflorenic were detected in glandular trichomes through the glandular microsampling technique and GC/MS analysis. In the glandular trichomes of the leaves also up to twenty-six volatile terpenes were identified, where beta-eudesmol and valencene were the most abundant terpenes.

Pro-ejaculatory effect of the aqueous crude extract of cihuapatli (Montanoa tomentosa) in spinal male rats.

In the present study, the pro-sexual effect of the cihuapatli (Montanoa tomentosa) and its possible pro-ejaculatory properties in spinal male rats were examined. Systemic administration of the aqueous crude extracts of Montanoa tomentosa exerted a pro-ejaculatory effect and produced an increase in the number of discharges in the ejaculatory motor patterns in the spinal rats. The cihuapatli-induced ejaculatory responses included the expression of penile erections and penile movements and the potent expulsion of urethral contents and in some cases the expulsion of seminal plugs. The cihuapatli-induced ejaculatory motor patterns were similar to that obtained after systemic oxytocin. Cihuapatli- and oxytocin-induced ejaculatory motor responses and the penile erections and movements were abolished by the pre-treatment with hexamethonium, a selective oxytocin antagonist. Present data show that the cihuapatli extract acts directly at the spinal system in charge of the expression of the ejaculatory motor patterns and suggest that the aqueous crude extract exerts its aphrodisiacs properties by increasing sexual potency acting as an oxytocic agent.

Photoperiodic induction of synchronous flowering near the Equator.

In tropical rainforests, 30-65% of tree species grow at densities of less than one individual per hectare. At these low population densities, successful cross-pollination relies on synchronous flowering. In rainforests with low climatic seasonality, photoperiodic control is the only reliable mechanism for inducing synchronous flowering. This poses a problem because there is no variation in day length at the Equator. Here we propose a new mechanism of photoperiodic timekeeping based on the perception of variation in sunrise or sunset time, which explains and predicts the annually repeated, staggered, synchronous and bimodal flowering of many tree species in Amazonian rainforests near the Equator.

Aphrodisiac properties of Montanoa tomentosa aqueous crude extract in male rats.

Cihuapatli, the Mexican zoapatle (Montanoa tomentosa) has an extensive ethnomedical history of use as a traditional remedy for reproductive impairments. During the study of the ejaculatory function in rats and by testing a set of Mexican plants with medicinal properties, we observed that crude extracts of M. tomentosa facilitated ejaculation. Thus, we decided to analyze the possibility that this plant possessed sexual stimulant properties. To that aim, copulatory behavior of sexually active male rats receiving doses of 38, 75 and 150 mg/kg of the aqueous crude extract of M. tomentosa, as it is prepared in traditional medicine, was assessed. In addition, we evaluated the effect of the 75-mg/kg dose of the extract on males with anesthetization of the genital area and on sexual behavior of sexually inactive male rats (noncopulators). Results showed that acute oral administration of crude extracts of M. tomentosa facilitates expression of sexual behavior in sexually active male rats, significantly increases mounting behavior in genitally anesthetized animals and induces the expression of sexual behavior in noncopulating males. Altogether, these data reveal a facilitatory action of this extract on sexual activity and particularly on sexual arousal. Present findings provide experimental evidence that the crude extract preparation of M. tomentosa, used as a traditional remedy, possesses aphrodisiac properties.

Sesquiterpene lactones from Montanoa hibiscifolia that inhibit the transcription factor NF-kappa B.

The reinvestigation of the aerial parts of Montanoa hibiscifolia afforded four new eudesmanolides (1-4), three of them with a rare endoperoxide structural element and the fourth with a rare carbonyl function. It also afforded three unusual montabibisciolides (5-7), two (5 and 7) of which are new natural compounds. Additionally, seven germacrolides (8-10 and 12-15) and one melampolide (11) could be isolated, including two new germacrolides (8 and 9). Their structures were elucidated using 1D and 2D NMR measurement as well as ESI, CIMS, and HRMS analyses. Low-energy conformations were obtained by molecular mechanics calculations. The (13)C NMR data of five compounds are reported for the first time. Six sesquiterpene lactones (4, 6, 10, 11, 12, and 14) were investigated for their inhibitory activity on DNA binding of the transcription factor NF-kappa B using Jurkat T cells as well as RAW 264.7 cells. Besides the alpha-methylene-gamma-butyrolactone moiety the epoxy group in the acyl residue might take part in the NF-kappa B inhibitory activity of sesquiterpene lactones.

Two new sesquiterpene lactones from Montanoa tomentosa ssp. microcephala.

Two new sesquiterpene lactones: 8alpha-(4'-acetoxymethacryloyloxy)-3alpha,9beta-dihydroxy-1(10)E,4Z,11(13)-germacratrien-12,6alpha-olide (1) and 8alpha-(2'E)-(2'-acetoxymethyl-2'-butenoyloxy)-3alpha,9beta-dihydroxy-1(10)E,4Z,11(13)-germacratrien-12,6alphaolide (2), together with the known zoapatanolide A were isolated from the aerial parts of Montanoa tomentosa Cerv. in La Llave et Lex ssp. microcephala (Sch. Bip. In K. Koch) V.A. Funk (Asteraceae). The structures of all compounds were established on the basis of 1D, 2D NMR, and EIMS analysis.

The zoapatle II--botanical and ecological determinants.

A collection of Montanoa (Cerv) specimens was conducted throughout Mexico. Twenty-one specimens were classified, some of them grown in the greenhouse and transplanted in an agricultural experimental field station in the Valley of Mexico. In vitro uterotonic potency was assayed and the results expressed as equivalents of oxytocine, by using estrogenized guinea pig uterine strips. A great variation of uterotonic potency was observed among the wild plants. A clear decrease and uniformity of uterotonic potency was found in plants grown in the experimental field. The study points out the importance of ecological variations in expression of the plant's biological activity.

PIP: 21 specimens of the zoapatle Montanoa (Cerv) plant were collected from sites throughout Mexico with a variety of climatic, humidity, soil composition, and altitude conditions to assess their uterotonic properties. Some plants were then grown in an agricultural experimental field station of the National Institute of Agricultural Research, where all the variables were known, allowing for a valid comparison between species and plants. In vitro uterotonic potency was assayed and the results expressed as equivalents of oxytocin. The wild plants were found to vary greatly in uterotonic potency, perhaps reflecting differences in ecological conditions. The most active uterotonic zoapatle aqueous crude extract was the Montanoa frutescens, and the least active was M. mollissima, the former having approximately 4 times the uterotonic potency of the latter. The plants grown in the agricultural field station showed a considerable decrease in uterotonic potency, but valid comparisons must await assay in winter to match the stage in the plant life when the wild samples were assayed. Plant age, the part of the plant from which the uterotonic principle is extracted, climate, altitude, and soil constituents are other possible influences on uterotonic potency. A sample of the zoapatle aqueous crude extract with the highest uterotonic potency will be assayed for its luteolytic activity in an animal model.

The zoapatle I--a traditional remedy from Mexico emerges to modern times.

A review of publications from 1529 to 1982 related to the zoapatle was made. Historical, biological, ethnobotanical, agricultural, clinical and chemical studies were noted; most of the early publications were written by Mexican scientists, in Spanish. More recent ones appear in the form of patents dealing with the isolation, characterization and total synthesis of some of its constituents. From this review, ideas emerged that might contribute to the understanding and rational use of this plant, or its chemical constituents, as a new, safe, reliable, inexpensive and - hopefully soon - a widely used interceptive agent.

PIP: Zoapatle, the common name for Montanoa tomentosa, is a well-known plant in Mexico that has been used since the 16th century to induce labor or to increase both the tone and frequency of uterine contractions during labor. Since 1866, the plant has been recognized as a potential menses inducer. Pharmacological observations of zoapatle were made in Mexico in 1945 and focused on the administration of zoapatle aqueous crude extract (ZACE). Animal studies demonstrated increased uterine contractility. 9 compounds have been isolated from zoapatle. Much of the recent work with the material has been detailed in several patent applications. In 2 of 4 clinical studies on the use of ZACE, an increase in uterine contractility and cervical dilation among pregnant women was noted, and in another study, a cramping sensation was reported by nonpregnant women. The need to control for ecological variables which effect the plants seasonally was mentioned by 1 researcher who found working with the crude extract difficult and unpredictable. The lack of toxic effects was demonstrated in a study using animal models. Different effects were noted in a study which used zoapatanol obtained by total synthesis, including the lack of uterotonic response in rabbit tissue which warrants further investigation. The plant or its derivatives are potentially useful as interceptive agents but further research under controlled conditions is needed.