Phytochemical Constituents, Antioxidant, Cytotoxic, and Antimicrobial Activities of the Ethanolic Extract of Mexican Brown Propolis.

Propolis is a complex mixture of natural sticky and resinous components produced by honeybees from living plant exudates. Globally, research has been dedicated to studying the biological properties and chemical composition of propolis from various geographical and climatic regions. However, the chemical data and biological properties of Mexican brown propolis are scant. The antioxidant activity of the ethanolic extract of propolis (EEP) sample collected in México and the isolated compounds is described. Cytotoxic activity was evaluated in a central nervous system and cervical cancer cell lines. Cytotoxicity of EEP was evaluated in a C6 cell line and cervical cancer (HeLa, SiHa, and CasKi) measured by the 3-(3,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium (MTT) assay. The antibacterial activity was tested using the minimum inhibitory concentration (MIC) assay. Twelve known compounds were isolated and identified by nuclear magnetic resonance spectroscopy (NMR). Additionally, forty volatile compounds were identified by means of headspace-solid phase microextraction with gas chromatography and mass spectrometry time of flight analysis (HS-SPME/GC-MS-TOF). The main volatile compounds detected include nonanal (18.82%), α-pinene (12.45%), neryl alcohol (10.13%), and α-pinene (8.04%). EEP showed an anti-proliferative effect on glioma cells better than temozolomide, also decreased proliferation and viability in cervical cancer cells, but its effectiveness was lower compared to cisplatin.

Effect of Pinocembrin Isolated from Mexican Brown Propolis on Diabetic Nephropathy.

Propolis is a resinous beehive product that has been used worldwide in traditional medicine to prevent and treat colds, wounds, rheumatism, heart disease and diabetes. Diabetic nephropathy is the final stage of renal complications caused by diabetes and for its treatment there are few alternatives. The present study aimed to determine the chemical composition of three propolis samples collected in Chihuahua, Durango and Zacatecas and to evaluate the effect of pinocembrin in a model of diabetic nephropathy in vivo. Previous research demonstrated that propolis of Chihuahua possesses hypoglycemic and antioxidant activities. Two different schemes were assessed, preventive (before renal damage) and corrective (once renal damage is established). In the preventive scheme, pinocembrin treatment avoids death of the rats, improves lipid profile, glomerular filtration rate, urinary protein, avoid increases in urinary biomarkers, oxidative stress and glomerular basement membrane thickness. Whereas, in the corrective scheme, pinocembrin only improves lipid profile without showing improvement in any other parameters, even pinocembrin exacerbated the damage. In conclusion, pinocembrin ameliorates diabetic nephropathy when there is no kidney damage but when it is already present, pinocembrin accelerates kidney damage.

In vitro antileishmanial activity of Mexican medicinal plants.

AIM OF THE STUDY: To evaluate the anti-leishmanial activity and cytotoxicity of aqueous and organic extracts of ten plants used in Mexican traditional medicine as anti-parasitics. MATERIALS AND METHODS: For the organic extracts, plant material was macerated in dichloromethane (CH2Cl2) and dichloromethane/methanol (CH2Cl2/MeOH) (1:1) during two weeks; the aqueous extracts were prepared by infusion. The extracts were tested against promastigotes and intracellular amastigotes of Leishmania amazonensis. The cytotoxicity was assayed in parallel on peritoneal macrophages of BALB/c mice. RESULTS: Four of the thirty extracts tested were active and selective against L. amazonensis promastigotes: Schinus molle (CH2Cl2 and CH2Cl2/MeOH), Lantana camara (CH2Cl2) and Prosopis laevigata (aqueous). These extracts had a median inhibitory concentration (IC50) against intracellular amastigotes under 50 µg/mL and a selectivity index (SI) higher than 5, which indicates that they constitute valuable candidates to obtain secondary metabolites with leishmanicidal activity. CONCLUSIONS: The results derived from this study indicate that L. camara, P. laevigata, and S. molle might provide interesting new leads for the development of antileishmanial drugs.

Analysis of volatile components from Melipona beecheii geopropolis from Southeast Mexico by headspace solid-phase microextraction.

A head space solid-phase microextraction method combined with gas chromatography-mass spectrometry was developed and optimised to extract and analyse volatile compounds of Melipona beecheii geopropolis. Seventy-three constituents were identified using this technique in the sample of geopropolis collected. The main compounds detected include ß-fenchene (14.53-15.45%), styrene (8.72-9.98%), benzaldehyde (7.44-7.82%) and the most relevant volatile components presents at high level in the geopropolis were terpenoids (58.17%).

Mexican propolis flavonoids affect photosynthesis and seedling growth.

As a continuous effort to find new natural products with potential herbicide activity, flavonoids acacetin (1), chrysin (2) and 4',7-dimethylnarangenin (3) were isolated from a propolis sample collected in the rural area of Mexico City and their effects on the photosynthesis light reactions and on the growth of Lolium perenne, Echinochloa crus-galli and Physalis ixocarpa seedlings were investigated. Acacetin (1) acted as an uncoupler by enhancing the electron transport under basal and phosphorylating conditions and the Mg(2+)-ATPase. Chrysin (2) at low concentrations behaved as an uncoupler and at concentrations up to 100 µM its behavior was as a Hill reaction inhibitor. Finally, 4',7-dimethylnarangenin (3) in a concentration-dependent manner behaved as a Hill reaction inhibitor. Flavonoids 2 and 3 inhibited the uncoupled photosystem II reaction measured from water to 2,5-dichloro-1,4-benzoquinone (DCBQ), and they did not inhibit the uncoupled partial reactions measured from water to sodium silicomolybdate (SiMo) and from diphenylcarbazide (DPC) to diclorophenol indophenol (DCPIP). These results indicated that chrysin and 4',7-dimethylnarangenin inhibited the acceptor side of PS II. The results were corroborated with fluorescence of chlorophyll a measurements. Flavonoids also showed activity on the growth of seedlings of Lolium perenne and Echinochloa crus-galli.

Ethnobotanical survey and antibacterial activity of plants used in the Altiplane region of Mexico for the treatment of oral cavity infections.

ETHNOPHARMACOLOGICAL RELEVANCE: The ethnobotanical survey conducted in this study showed 47 plant species used in the Altiplane region of Mexico for the treatment of dental diseases such as toothache, dental caries, periodontal disease and gingivitis. MATERIALS AND METHODS: Information was collected by performing interviews. The following data were recorded: name of the patients or herbalists with their age, sex; date and place of gathering information; pathology of persons interviewed; name of the drug (vernacular name); parts used (leaves, bark, fruits, seeds, aerial parts), mode of preparation and administration, and possible combinations. The inhibitory effects of the aqueous and ethanolic extracts of the medicinal plants detected during the survey the on the growth of Streptococcus mutans and Phrophyromonas gingivalis were determined using microdilution method. The minimum bactericidal concentrations (MBC) were determined from the wells of microplate with no visible bacterial growth. RESULTS: In total, tree places of the Altiplane region of Mexico were visited and five healers and 100 patients were questioned. 47 wild and cultivated species were recorded. The most frequent uses were to treat tooth pain, gum diseases, bad breath and cavities. Infusions were the most frequently prepared formulation. Other applied preparations mentioned with decreasing frequency were decocts, syrups, tinctures, direct application of the plant material without prior preparation and finally macerations. The ethanolic extracts of Haematoxylon brasiletto, Punica granatum, Iostephane heterophyla, Bursera simaruba, Cedrela odorata and Rhus standleyi (12.5-65.0 µg/mL) as well as water extracts of Haematoxylon brasiletto, Punica granatum, Iostephane heterophyla, Amphipterygium adstringens, Argemone mexicana, Cedrela odorata, Eysenhardtia polystachya, Persea americana, Syzygium aromaticum, Cinnamomun zeylanicum, Cnidoscolus multilobus and Rhus standleyi (10.5-78.0 µg/mL) showed the highest inhibitory effect against Streptococcus mutans and Porphyromonas gingivalis. CONCLUSIONS: Many plants are used in the Mexican traditional medicine to treat oral bacterial diseases by the healers or patients. Our study demonstrated that most of the medicinal plants showed an antibacterial effect in vitro, and justified at least in part their use in traditional medicine. These results encourage further investigations to extract and identify the active chemical compounds responsible for the antibacterial effect observed.

Ent-trachyloban-19-oic acid isolated from Iostephane heterophylla as a promising antibacterial agent against Streptococcus mutans biofilms.

From the roots of Iostephane heterophylla, six known compounds, namely, ent-trachyloban-19-oic acid (1), the mixture of ent-kaur-16-en-19-oic acid (2) and ent-beyer-15-en-19-oic acid (3), xanthorrhizol (4), 16α-hydroxy-ent-kaurane (5) and 16α-hydroxy-ent-kaur-11-en-19-oic acid (6) were isolated using a bioassay-guided fractionation method. The known compounds (1-6) were identified by comparison of their spectroscopic data with reported values in the literature. In an attempt to increase the resultant antimicrobial activity of 1 and 4, a series of reactions was performed on ent-trachyloban-19-oic acid (1) and xanthorrhizol (4), to obtain derivatives 1a, 1b, and 4a-4d. All the isolated compounds (1-6) and the derivatives 1a, 1b, and 4a-4d were evaluated for their antimicrobial activity against two oral pathogens, Streptococcus mutans and Porphyromonas gingivalis associated with caries and periodontal disease, respectively. Compounds 1, 1b, 2+3, 4 and 4d inhibited the growth of S. mutans with concentrations ranging from 4.1 µg/mL to 70.5 µg/mL. No significant activity was found on P. gingivalis except for 4 with an MIC of 6.8 µg/mL. The ability of 1, 1b, 2+3, 4 and 4d to inhibit biofilm formation by S. mutans was evaluated. It was found that 1, 1b, 4 and 4d interfered with the establishment of S. mutans biofilms, inhibiting their development at 32.5, 125.0, 14.1 and 24.4 µg/mL, respectively.

Antimicrobial compounds isolated from Haematoxylon brasiletto.

ETHNOPHARMACOLOGICAL RELEVANCE: The decoction of the bark of Haematoxylon brasiletto Karst. (Leguminosae), commonly known as "Palo Brasil", is used in the Mexican traditional medicine to treat mouth and kidney infections, hypertension, stomach upsets, gastric ulcers and diabetes. AIM OF THE STUDY: The present study was performed to evaluate the antimicrobial effects of the methanolic extract of the bark of Haematoxylon brasiletto. MATERIALS AND METHODS: A panel of 12 bacteria and the yeast Candida albicans were used. Minimum inhibitory concentrations (MICs) were determined by the standard broth microdilution method. RESULTS: The results indicate that the extract of Haematoxylon brasiletto inhibited the growth of eight of the tested microorganisms at a concentration limit of 128 microg/mL. For the tested compounds the MIC values ranged from 8.7 to 128 microg/mL. CONCLUSIONS: The overall results provide promising baseline information for the potential use of the extracts from Haematoxylon brasiletto as well as some of the isolated compounds in the treatment of bacterial infections.