Variation in the Floral Morphology of Prosthechea karwinskii (Orchidaceae), a Mexican Endemic Orchid at Risk.

Prosthechea karwinskii is an orchid endemic to Mexico, threatened by the destruction of its habitat and the extraction of specimens to meet its demand for ornamental and religious use. Most of its populations, including the most locally abundant ones, are found in Oaxaca state. Variations in some floral traits have been observed in these populations. We implemented a morphometric analysis to assess their floral variation and identify the most significant characters in the morphological patterns of this orchid. Floral samples were collected from 17 populations of P. karwinskii in Oaxaca, as well as from specimens used as ornaments during Easter in an Oaxacan community (Zaachila), whose origin is unknown. Sampling of natural populations covered the environmental, geographic, and morphological variation of the species. We performed an analysis of variance (ANOVA), principal component analysis (PCA), canonical variate analysis (CVA), and cluster analysis, including 185 individuals and 45 variables (12 of them were discarded in the multivariate analyses due to high correlation). Characters of the column, lateral sepal, and labellum were most informative for the observed morphological patterns. Albarradas showed the greatest morphological differentiation, mainly due to the column. In general, individuals from the same locality tended to overlap more, especially the populations of Jaltianguis and Yahuiche, which were different from the geographically close population of Etla. Teposcolula presented the highest values in perianth characters, unlike Sola_Rancho Viejo. The specimens recovered from religious ornaments were morphologically more similar to those from Yanhuitlan and Etla. This morphometric analysis identified characters as potential taxonomic markers for P. karwinskii and related species, showing its potential to associate specimens of unknown origin with their probable geographical region. Our work encourages working on collaborative conservation strategies to ensure the long-term permanence of both the species and its traditional uses.

Medicinal Orchids of Mexico: A Review.

Some species of the Orchidaceae family are used in Mexican traditional medicine. However, there are no current and critical compilations of the medicinal uses and pharmacological effects of the members of the Orchidaceae family. This review provides a current, critical, and comprehensive analysis of the traditional medicinal uses, pharmacological reports, and active compounds isolated from Mexican orchids. A total of 62 Mexican orchids with medicinal potential have been recorded, of which 14 have scientific evidence. The remaining 48 plant species have ethnomedicinal information but have not been validated with scientific studies. These orchids are distributed in 14 states of the Mexican Republic, mainly in the southern region of Mexico. The most common pharmacological activities reported are anti-inflammatory, vasorelaxant, antinociceptive, antioxidant, spasmolytic, antihypertensive, and hallucinogenic activities. It is necessary to increase the number of pharmacological, phytochemical, and toxicological studies with medicinal orchids from Mexico because there are scientific studies on only 22.5% of these species. In further studies, it will be possible to evaluate the pharmacological effects of Mexican orchids in clinical trials. In addition, the mechanisms of action by which plant extracts and their active compounds exert medicinal effects remain to be studied. Plant extracts from orchids and their active compounds show promising antinociceptive and spasmolytic effects, respectively.

Composition of essential oil from the bark and leaves of two morphotypes of Salmea scandens (Asteraceae), a medicinal plant in Oaxaca, Mexico.

The chemical composition of the essential oils (EOs) from leaves and bark of two morphotypes, white (WM) and black (BM) of Salmea scandens obtained by microwave-assisted extraction (MAE) were analyzed by GC-MS. The major components of the bark EOs were aliphatic hydrocarbons (38.0% in WM, 48.6% in BM) and oxygenated sesquiterpenes (27.6% in WM, 11.3% in BM); in leaves EOs major components were oxygenated sesquiterpenes (43.9% in WM, 45.7% in BM) and oxygenated aliphatics (13.7% in WM, 1.1% in BM) stood out. Nine components have been reported to have antimicrobial and anti-inflammatory potential. The agglomerative hierarchical clustering and principal component analysis confirmed the variability of the EOs. These results suggest that WM may have a better therapeutic effect for treating infectious and inflammatory processes in traditional medicine.

Microwave-Assisted Hydrodistillation of Essential Oil from Plectranthus amboinicus: Evaluation of Its Antifungal Effect and Chemical Composition.

Fusarium wilt, a vascular syndrome in a wide range of plants, is caused by the pathogen Fusarium Oxysporum. The objective of this investigation was to evaluate the antifungal effect of four essential oils (EOs) (Plectranthus amboinicus, Syzygium aromaticum, Lippia alba, and Rosmarinus officinalis), which were obtained by using microwave-assisted hydrodistillation (MAH), against F. oxysporum. The yield obtained from P. amboinicus with the use of MAH was 0.2%, which was higher than that of a conventional extraction; its extraction time was also shorter. For concentrations of 100 and 300 µL/L, P. amboinicus caused an inhibition rate of 27.2 and 55.7%, respectively, while S. aromaticum caused an inhibition rate of 23.1 and 87.3%, respectively. It was observed that increasing the concentration also increased the % inhibition rate. The extracts of L. alba and R. officinalis caused an inhibition rate of 14.5 and 14.7% at 500 µL/L, respectively, at 10 days of incubation, while at this concentration, P. amboinicus and S. aromaticum achieved 100%. The major chemical compounds of P. amboinicus were carvacrol (41.20%), o-cymene (11.61%), caryophyllene (11.45%), α-bergamotene (7.71%), and caryophyllene oxide (4.62%), and these monoterpene hydrocarbons were responsible for the biological activity. The essential oil of P. amboinicus in appropriate concentrations is a potent antifungal agent that could be used for the control of F. oxysporum.

Effect of Microwave and Ultrasound during the Killing Stage of the Curing Process of Vanilla (Vanilla planifolia, Andrews) Pods.

The curing process (CP) of Vanilla planifolia pods, which is a long and tedious process, is necessary to obtain the natural vanilla extract. This research evaluated the application of microwave (M) and ultrasound (U) during the "killing" stage of the CP and its effect on vanillin content and ß-glucosidase activity. The pods were immersed in a container with water or with moistened samples for the M treatments. In U treatments, the pods were immersed in an ultrasonic bath. After this stage, the samples were subjected to an additional U treatment. The results show that the application of these technologies significantly improves vanillin yield (p < 0.05) and the curing time is reduced to 20 days. U treatments subjected to additional sonication at 38 °C obtain more than double the yield of vanillin regarding control. The effect of M and U on cell structure damage increases with additional sonication, but at 15 min, ß-glucosidase inactivation decreases the final yield. Disposition of samples in M also affects the final vanillin content. There is no significant correlation between ß-glucosidase and vanillin in the different treatments. The application of M and U with the appropriate parameters reduces the CP time without affecting the compounds of interest.

Role of LTB4 and nitric oxide in the gastroprotective effect of Prosthechea karwinskii leaves extract in the indomethacin-induced gastric injury in the rat.

Gastric injury is mainly described by inflammation of the gastric epithelium. Recently, our group of work demonstrated that Prosthechea karwinskii leaves extract induces both an in vitro antioxidative action and an in vivo gastroprotective effect in a rat. However, the molecules involved in the gastroprotective action by Prosthechea karwinskii are not known. Thus, the aim of this study is to determine whether Prosthechea karwinskii extract modifies anti-inflammatory and antioxidative biomarkers in an in vivo rat model of indomethacin-induced gastric injury. Rats were orally administered with indomethacin and Prosthechea karwinskii leaf extract. Our results suggest that the gastroprotective effect of Prosthechea karwinskii leaf extract is related to the reduction in leukocyte infiltration and antioxidative action in a model of indomethacin-induced gastric injury. Further studies are warranted to investigate the role of the compounds identified in the gastroprotective action of Prosthechea karwinskii leaves extract.

Validation of the traditional medicinal use of a Mexican endemic orchid (Prosthechea karwinskii) through UPLC-ESI-qTOF-MS/MS characterization of its bioactive compounds.

Ethnopharmacological relevance: The orchid Prosthechea karwinskii is a medicinal orchid in Oaxaca, Mexico, used to treat diabetes, cough, wounds, and burns, prevent miscarriage and assist in labor. Each part of the plant (leaves, pseudobulbs, or flowers) is used by healers for certain treatment conditions, indicating that each part has different biocompounds with specific pharmacological activity. Aim of the study: To characterize the biocompounds in extracts from leaves, pseudobulbs, and flowers of P. karwinskii and evaluate their ROS inhibition capacity to associate it with medicinal uses. Materials and methods: The compounds present in extracts from leaves, pseudobulbs, and flowers of P. karwinskii were identified by UPLC-ESI-qTOF-MS/MS. The chemical differentiation of each extract was tested by principal component analysis (PCA) using compound intensity values. For each extract, total phenol and flavonoid contents were quantified. Their antioxidant capacity was evaluated ex vivo by inhibition of ROS with DCFH-DA and in vitro with DPPH radical. Results: Based on the PCA, it was observed that some compounds were completely separated from others according to the correlation that they presented. The compounds common to all three plant parts were quinic, malic, succinic, azelaic, and pinellic acids. Among the compounds identified, two were exclusive to leaves, four to pseudobulbs, and ten to flowers. Some of the identified compounds have well-known antioxidant activity. The leaves had the highest content of total phenols and flavonoids, and the highest in vitro and ex vivo antioxidant capacity. A strong correlation was observed between phenol and flavonoid contents, and antioxidant capacity ex vivo and in vitro. Conclusions: It was found that the bioactive compounds and antioxidant capacity of each part of the plant were associated with its traditional medicinal use. A pharmacological potential was also found in P. karwinskii for further biological studies because of the type of compounds it contained.

Bioactive compounds from Prosthechea karwinskii decrease obesity, insulin resistance, pro-inflammatory status, and cardiovascular risk in Wistar rats with metabolic syndrome.

ETHNOPHARMACOLOGICAL RELEVANCE: The orchid Prosthechea karwinskii is a species endemic to Mexico, which is used in traditional medicine to lower glucose levels in patients with diabetes, and to treat inflammation-related problems. Recent studies have shown that this orchids can reduce glucose, cholesterol, and triglyceride levels in Wistar rats, which were previously induced to have metabolic syndrome (MS). AIM OF THE STUDY: To evaluate the effect of P. karwinskii leaves extract on the components of metabolic syndrome: obesity, insulin resistance, pro-inflammatory status, and cardiovascular risk in a Wistar rat model, and to identify the bioactive compounds in the extract. MATERIALS AND METHODS: UPLC-ESI-qTOF-MS/MS was used to identify the compounds present in the extract. MS was induced in Wistar rats through administration of a 40% sucrose diet for 20 weeks. The rats were divided into five groups that received different treatments for 4 weeks: one group without any treatment, one group receiving metformin (200 mg/kg p.o.), and three groups receiving different doses of P. karwinskii leaves extract (100, 200, and 300 mg/kg p.o.). The animals' body weights were recorded weekly, and at the end of the experiment, they were sacrificed after fasting for 18 h to determine the levels of glucose, insulin, insulin resistance index, total cholesterol, triglycerides, and adiponectin in the serum, as well as levels of TNF-α and HS-CRP in the serum and liver homogenates. The abdominal and pericardial fat weights were also recorded. RESULTS: The main bioactive compounds of the extract were quinic acid, neochlorogenic acid, chlorogenic acid, rutin, kaempferol-3-o-ß-rutinoside, and embelin, known to exhibit MS-related bioactivity. Oral administration of P. karwinskii leaves extract at a dose of 300 mg/kg decreased weight gain, abdominal and pericardial fat deposits, and insulin resistance. At the end of the treatment, levels of triglycerides, TNF-α, HS-CRP, and adiponectin returned to levels similar to normal. CONCLUSION: P. karwinskii extract (300 mg/kg) had an anti-obesity effect, decreased insulin resistance, pro-inflammatory status, and cardiovascular risk in rats with induced MS by increasing adiponectin levels and decreasing TNF-α and HS-CRP levels. The compounds identified in the extract could be responsible for these effects, acting alone or in synergy, as several compounds in the extract are known to have MS-related bioactivity. The foliar extract of P. karwinskii has potential as an effective alternative to a cocktail of drugs used to treat problems associated with MS.

Prosthechea karwinskii, an orchid used as traditional medicine, exerts anti-inflammatory activity and inhibits ROS.

ETHNOPHARMACOLOGICAL RELEVANCE: Prosthechea karwinskii (Mart.) J.M.H. Shaw is a Mexican orchid used in traditional medicine by some indigenous communities to treat issues related to inflammation (cough, wounds, burns, and diabetes). Pharmacological research of this orchid could validate its therapeutic uses and demonstrate its potential for treating other health conditions of high prevalence in Mexico, including those associated with oxidative stress such as diabetes, cancer, atherosclerosis, and hypertension as well as inflammation. AIM OF THE STUDY: The leaf extract from P. karwinskii was examined to identify its compounds and elucidate its inhibitory effect on reactive oxygen species as well as its anti-inflammatory activity and gastroprotective effects in an animal model. MATERIALS AND METHODS: Compounds were identified via ultra-high-performance liquid chromatography coupled with electrospray ionization with quadrupole time of flight-mass spectrometry. Inhibition of reactive oxygen species was determined ex vivo in peripheral blood mononuclear cells with 2',7'-dichlorodihydrofluorescein diacetate. The anti-inflammatory activity was assessed using a carrageenan-induced paw edema model in Wistar rats; nitric oxide and tumor necrosis factor alpha levels were quantified. The gastroprotective effect was evaluated in Wistar rats with indomethacin-induced gastric injury. RESULTS: Nine compounds were identified in the P. karwinskii leaf extract. Most compounds, such as quinic acid, malic acid, neochlorogenic acid, chlorogenic acid, rutin, embelin, pinellic acid, and azelaic acid, were reported to exhibit antioxidant and/or anti-inflammatory activity. The extract was also found to inhibit reactive oxygen species in the ex vivo model. Unlike other anti-inflammatory drugs, the extract exerted a dual effect: anti-inflammatory activity and protection of the gastric mucosa. The results showed that the extract could significantly inhibit the release of nitric oxide without a dose-response relationship. CONCLUSION: P. karwinskii leaf extract inhibited reactive oxygen species and exerted an anti-inflammatory effect. Moreover, this extract did not induce gastric damage in the animals. The bioactivity of the species was found to support its use in traditional medicine. This orchid could be used to treat inflammatory diseases without causing the side effects associated with nonsteroidal anti-inflammatory drugs. It can also be employed to treat other pathological conditions associated with oxidative stress. The findings herein form the basis for the future discovery of natural products that may serve as safe alternative therapies for inflammatory disorders.