Ethnopharmacological Use Research Articles

Anxiolytic properties of Cymbopogon citratus (DC.) stapf extract, essential oil and its constituents in zebrafish (Danio rerio)

Ethnopharmacological relevanceCymbopogon citratus (DC.) Stapf (Poaceae) leaves is often consumed as infusion in folk medicine due to its therapeutic properties. This plant is also rich in essential oil, which has several beneficial effects to the human health. It is known that medications commonly used to treat anxiety disorders cause undesirable side effects. Thus, it is important to evaluate the anxiolytic effects of natural products from plants, such as C. citratus, as an alternative therapy to treat these disorders. Objective: The aim of this study was to investigate the anxiolytic properties of C. citratus essential oil (EO), hydroalcoholic extract (E1), citral (CIT), geraniol (GER) and the mixture of these terpenoids, as well as its possible mechanism of action by using zebrafish as an anxiety model. Methods: Adult zebrafish were treated (by immersion) with C. citratus EO, E1, CIT and/or GER. The anxiolytic effects were analyzed by using the light-dark test. The mechanism involved in the anxiolytic effects was further investigated by the coadministration of flumazenil (FMZ), an antagonist of GABAA receptors. The total polyphenols (phenolic and flavonoid compounds) content of E1 was determined by using spectrophotometric assays. Results: All analyzed samples showed a remarkable anxiolytic effect on zebrafish in the highest concentrations, as the animals showed a preference for the light side of the tank. Furthermore, the observed effect of EO, E1, CIT and GER was reversed by pre-treatment with FMZ, suggesting that GABAergic receptors were involved in the anxiolytic effect displayed by these samples. The association between CIT and GER in the lowest studied concentrations showed an interesting synergistic behavior on anxiolytic effect observed in light-dark test. Besides, it was demonstrated that E1 was constituted by phenolic and flavonoid compounds, which could be involved in the observed effect. Conclusion: This work has proved that the low-cost zebrafish can be an adequate alternative as an animal model to evaluate the anxiolytic effect of C. citratus and its related compounds. Moreover, the involvement of GABAA receptors could be responsible for the effect showed by the samples. These obtained results can potentially validate the ethnopharmacological use of C. citratus as a medicinal plant for the treatment of anxiety disorders in folk medicine.

Antiseizure potential of the ancient Greek medicinal plant Helleborus odorus subsp. cyclophyllus and identification of its main active principles

Ethnopharmacological relevanceEthnopharmacological data and ancient texts support the use of black hellebore (Helleborus odorus subsp. cyclophyllus, Ranunculaceae) for the management and treatment of epilepsy in ancient Greece. Aim of the studyA pharmacological investigation of the root methanolic extract (RME) was conducted using the zebrafish epilepsy model to isolate and identify the compounds responsible for a potential antiseizure activity and to provide evidence of its historical use. In addition, a comprehensive metabolite profiling of this studied species was proposed. Materials and methodsThe roots were extracted by solvents of increasing polarity and root decoction (RDE) was also prepared. The extracts were evaluated for antiseizure activity using a larval zebrafish epilepsy model with pentylenetetrazole (PTZ)-induced seizures. The RME exhibited the highest antiseizure activity and was therefore selected for bioactivity-guided fractionation. Isolated compounds were fully characterized by NMR and high-resolution tandem mass spectrometry (HRMS/MS). The UHPLC-HRMS/MS analyses of the RME and RDE were used for dereplication and metabolite profiling. ResultsThe RME showed 80% inhibition of PTZ-induced locomotor activity (300 μg/ml). This extract was fractionated and resulted in the isolation of a new glucopyranosyl-deoxyribonolactone (1) and a new furostanol saponin derivative (2), as well as of 20-hydroxyecdysone (3), hellebrin (4), a spirostanol glycoside derivative (5) and deglucohellebrin (6). The antiseizure activity of RME was found to be mainly due to the new furostanol saponin (2) and hellebrin (4), which reduced 45% and 60% of PTZ-induced seizures (135 μM, respectively). Besides, the aglycone of hellebrin, hellebrigenin (S34), was also active (45% at 7 μM). To further characterize the chemical composition of both RME and RDE, 30 compounds (A7-33, A35–37) were annotated based on UHPLC-HRMS/MS metabolite profiling. This revealed the presence of additional bufadienolides, furostanols, and evidenced alkaloids. ConclusionsThis study is the first to identify the molecular basis of the ethnopharmacological use of black hellebore for the treatment of epilepsy. This was achieved using a microscale zebrafish epilepsy model to rapidly quantify in vivo antiseizure activity. The UHPLC-HRMS/MS profiling revealed the chemical diversity of the extracts and the presence of numerous bufadienolides, furostanols and ecdysteroids, also present in the decoction.

Anticoagulant active ingredients identification of total saponin extraction of different Panax medicinal plants based on grey relational analysis combined with UPLC-MS and molecular docking

Ethnopharmacological relevanceIn traditional use, Panax medicinal plants (ginseng, red ginseng, notoginseng, Panax japonicus, and Panacis majoris rhizoma) have different bioactivities from each other, even under different dosages, but their chemical compositions are very similar; so the question is, what is the primary effective substance induced the different efficacy, and how to identify them from a group of chemical constituents? Aim of the studyThe goal of this research was to provide a strategy to determine the effective substance in Panax genus medicinal plants responsible for the anticoagulant response. Materials and methodsThis research used ultra-high performance liquid chromatography coupled with mass spectrometry (UPLC-MS) to analyze the saponin chemical compositions of different concentration ethanol extraction of notoginseng and the ginseng genus medical plant 75% ethanol extraction (Panax ginseng, red ginseng, Panax japonicus, Panacis majoris rhizome), and used four coagulation factors (PT, TT, APTT, Fib) to evaluate the anticoagulant activity of the extracts. Grey correlation analysis was applied to establish the spectral effect relationship and give the anticoagulant potency of different saponins. Network pharmacology and molecular docking were adopted to clarify and verify the possible mechanisms of anticoagulant action. ResultsThe results showed that the blood physiological regulation activities of Panax medicinal plants were different according to the solvent concentration, processing, species and dosage. Overall, the most suitable solvent for extraction of SQ was 75% ethanol; At low dosage (10–100 mg/mL), the anticoagulant effect of Panax medical plants was: ZJS > ZZS > SQ > RS > HS, and at high doses (100–1000 mg/mL) was: SQ > ZJS > ZZS > RS > HS. GRA and molecular docking results showed the contribution of some components (NG-R2, NG-Fc/G-Ra1/G-Ra2, G-Rc, G-Rk3, and G-Rh4) to the whole anticoagulant activity of the drug were increased, while the effect of CS-IVa was just decreased with the increase of dosage; the anticoagulant effect of G-Rg3 (the main anticoagulant component) is mainly related to the targets F2, AR, RHO, ACR, MB, GZMB, B2M, CA2, CAT, and PAPOLA. ConclusionThis study determined the effective substance of anti-coagulation of ginseng genus herbal medicines and the regulation of different anticoagulant effects of TCM by changing various influencing conditions, including processing method, extraction method, and dose. It also provided an effective strategy for effective substances identification of multicomponent, multifunction, and multipurpose herbal medicine.

Ethnopharmacology, Pharmacology and Phytochemistry of Aristolochia bracteolata Lam: A Review of an Antimalarial Plant

Malaria remains one of the most common infectious diseases in the sub-Sahara African countries and other developing countries. Among the medicinal plants used in the endemic countries for the treatment of malaria is Aristolochia bracteolata Lam. due to its availability, accessibility, and traditional use. This study therefore reviewed the ethnomedicinal use, pharmacology, and the chemistry of Aristolochia bracteolata. Different electronic databases such as Medline/Pubmed, Cochrane Library, and Embase were searched to identify all published articles on Aristolochia bracteolata Lam. Key search words included ethnopharmacological use, pharmacological and phytochemical parameters of A. bracteolata. Retrieved articles were reviewed and synthesized. In addition, the reference list of retrieved articles was reviewed and articles which were not retrieved by previous search were hand searched. The review included original research articles that has investigated Aristolochia bracteolata Lam. of any study design. Only published original articles, any languages, any time of publish, and grey literature (Conference paper, theses both PhD. and Msc. technical report) were included. Those articles with full text not available, those without information of interest, e.g ethnopharmacology, pharmacology and phytochemistry of A. bracteolata were excluded. Despite having multiple use, the plant is mainly used in the treatment of malaria with a reported antiplasmodial activity. Aristolochic acids (AAs) were reported as the major and active ingredient among other components in the plant. The review revealed that A. bracteolata has various traditional use with promising pharmacological activity. However, information on its safety is limited.

In vitro and in vivo antitrypanosomal efficacy of combination therapy of Anogeissus leiocarpus, Khaya senegalensis and potash

Ethnopharmacological relevancePastoralists in Nigeria mix barks of Anogeissus leiocarpus (AL) Khaya senegalensis (KS) and potash (Pt) to treat animal African trypanosomosis. AimTo evaluate antitrypanosomal potential of A. leiocarpus, K. senegalensis and potash for insights into the traditional claim of antitrypanosomal combination therapy (ATCT). Materials and methodsFifty microliter each of six different concentrations of AL, KS, Pt, AL + KS, AL + KS + Pt and diminazene aceturate (DA, positive control) was incubated with 50 μL of parasite-laden blood containing 108Trypanosoma congolense cells in a 96-well microtitre plate. Negative control wells were devoid of the extracts and drug but supplemented with phosphate-buffered saline (PBS). Efficacy of treatment was observed at 1 h interval for complete immobilisation or reduced motility of the parasites. Each incubated mixture was inoculated into mouse at the point of complete immobilisation of parasite motility or at the end of 6-h observation period for concentrations that did not immobilise the parasites completely. For in vivo assessment, thirty-five parasitaemic rats were randomly allocated into seven groups of 5 rats each. Each rat in groups I–V was treated with 500 mg/kg of AL, KS, Pt, AL + KS and AL + KS + Pt, respectively, for 7 days. Rats in groups VI and VII were treated with diminazene aceturate 3.5 mg/kg once and PBS 2 mL/kg (7 days), which served as positive and negative controls, respectively. Daily monitoring of parasitaemia through the tail vein, packed cell volume and malondialdehyde were used to assess efficacy of the treatments. ResultsThe AL + KS + Pt group significantly (p < 0.05) and dose-dependently reduced parasite motility and completely immobilized the parasites at 10, 5 and 2.5 μg/μL with an IC50 of 9.1×10-4 µg/µL. All the mice with conditions that produced complete cessation of parasite motility did not develop parasitaemia within one month of observation. The AL + KS group significantly (p < 0.05) lowered the level of parasitaemia and MDA, and significantly (p < 0.05) maintained higher PCV than PBS group. ConclusionThe combination of A. leiocarpus and K. senegalensis showed better antitrypanosomal effects than single drug treatment and offers prospects for ATCT. Our findings support ethnopharmacological use of combined barks of A. leiocarpus and K. senegalensis by pastoralist in the treatment of animal African trypanosomosis in Nigeria.

Propolis extract has bioactivity on the wall and cell membrane of Candida albicans

Ethnopharmacological relevanceThe use of natural products such as propolis extract (PE) is a promising alternative when topically administered to replace conventional antifungals, mostly due to its therapeutic applications, ease of access and low toxicity. However, despite being the subject of several mycology studies, they focus primarily on exploiting their antimicrobial activity, lacking information on the mechanisms of action of PE on Candida spp., characterizing its antifungal potential. Aim of the studyTo elucidate the bioactivity of PE on the cellular structure of Candida albicans. Materials and methodsA total of seven C. albicans clinical isolates plus a reference strain of C. albicans ATCC 90028 were used in this study. The PE was characterized and its effect on C. albicans was determined by susceptibility and growth kinetics assays; interference on C. albicans germination and filamentation; evaluation of the integrity of the C. albicans cell wall and membrane, as well as its mutagenic potential. ResultsThe PE presented strong inhibitory activity, which showed its greatest antifungal activity at 12 h with dose and time dependent fungistatic characteristics, effectively inhibiting and interfering on C. albicans filamentation. In addition, PE caused membrane and cell wall damage with intracellular content extravasation. Moreover, PE was not mutagenic. ConclusionsThe bioactivity of PE is mainly related to the loss of integrity membrane as well as the integrity of the cell wall and consequent increase in permeability, without mutagenic effects.

Quantitative ethnopharmacological documentation and molecular confirmation of medicinal plants used by the Manobo tribe of Agusan del Sur, Philippines

BackgroundThe Philippines is renowned as one of the species-rich countries and culturally megadiverse in ethnicity around the globe. However, ethnopharmacological studies in the Philippines are still limited especially in the most numerous ethnic tribal populations in the southern part of the archipelago. This present study aims to document the traditional practices, medicinal plant use, and knowledge; to determine the relative importance, consensus, and the extent of all medicinal plants used; and to integrate molecular confirmation of uncertain species used by the Agusan Manobo in Mindanao, Philippines.MethodsQuantitative ethnopharmacological data were obtained using semi-structured interviews, group discussions, field observations, and guided field walks with a total of 335 key informants comprising of tribal chieftains, traditional healers, community elders, and Manobo members of the community with their medicinal plant knowledge. The use-report (UR), use categories (UC), use value (UV), cultural importance value (CIV), and use diversity (UD) were quantified and correlated. Other indices using fidelity level (FL), informant consensus factors (ICF), and Jaccard’s similarity index (JI) were also calculated. The key informants’ medicinal plant use knowledge and practices were statistically analyzed using descriptive and inferential statistics.ResultsThis study enumerated the ethnopharmacological use of 122 medicinal plant species, distributed among 108 genera and belonging to 51 families classified in 16 use categories. Integrative molecular approach confirmed 24 species with confusing species identity using multiple universal markers (ITS, matK, psbA-trnH, and trnL-F). There was strong agreement among the key informants regarding ethnopharmacological uses of plants, with ICF values ranging from 0.97 to 0.99, with the highest number of species (88) being used for the treatment of abnormal signs and symptoms (ASS). Seven species were reported with maximum fidelity level (100%) in seven use categories. The correlations of the five variables (UR, UC, UV, CIV, and UD) were significant (rs ≥ 0.69, p < 0.001), some being stronger than others. The degree of similarity of the three studied localities had JI ranged from 0.38 to 0.42, indicating species likeness among the tribal communities. Statistically, the medicinal plant knowledge among respondents was significantly different (p < 0.001) when grouped according to education, gender, social position, occupation, civil status, and age but not (p = 0.379) when grouped according to location. This study recorded the first quantitative ethnopharmacological documentation coupled with molecular confirmation of medicinal plants in Mindanao, Philippines, of which one medicinal plant species has never been studied pharmacologically to date.ConclusionDocumenting such traditional knowledge of medicinal plants and practices is highly essential for future management and conservation strategies of these plant genetic resources. This ethnopharmacological study will serve as a future reference not only for more systematic ethnopharmacological documentation but also for further pharmacological studies and drug discovery to improve public healthcare worldwide.

Comparison of Techniques and Solvents on the Antimicrobial and Antioxidant Potential of Extracts from Acacia dealbata and Olea europaea.

Ethnopharmacological use of plant natural extracts has been known since ancient times. The optimization of plant molecule extraction is fundamental in obtaining relevant extraction yields. The main purpose of this study was to understand the role of different extraction techniques (solid-liquid, ultrasound, Soxhlet, and microwave) and solvents (water, methanol, ethanol, acetone, dichloromethane, and hexane) on the antimicrobial and antioxidant activities of extracts from Olea europaea (olive) and Acacia dealbata (mimosa). Crude plant extracts were evaluated for their antimicrobial activity against Staphylococcus aureus and Escherichia coli by the disk diffusion method. The antioxidant capacity of the extracts was determined by ABTS (2,2-azinobis (3-ethyl-benzothiazoline-6-sulfonic acid)) and DPPH (2,2-diphenyl-1-picrylhydrazyl) methods. In terms of extraction yield, ultrasound extraction and the solvents methanol, acetone (O. europaea) or water (A. dealbata) were found to be the best options. However, ethanol and acetone proved to be the best solvents to extract compounds with antimicrobial activity and antioxidant capacity, respectively (regardless of the extraction method employed). Soxhlet and microwave were the best techniques to extract compounds with antimicrobial activity, whereas any of the tested techniques showed the ability to extract compounds with antioxidant capacity. In most of the cases, both plant extracts (mimosa and olive) were more efficient against S. aureus than E. coli. In the present study, both mimosa and olive leaf crude extracts proved to have antimicrobial and antioxidant activities, increasing the demand of these natural products as a source of compounds with health benefits.

Antimicrobial activities of Dillenia indica Linn. (Dilleniaceae) and Spondias mombin Linn. (Anacardiaceae) extracts on selected pathogenic organisms.

Background: The prevalence of increasing bacterial resistance against antibacterial agents in the past decade has spurred the search for substitutes. This necessitates the need to identify new compounds as alternatives to common synthetic antimicrobial compounds. Objectives: To establish and evaluate the antimicrobial potentials of Dillenia indica and Spondias mombin against strains of some selected pathogens. Method: Concentrations of the extract (methanol and dichloromethane) were prepared and the susceptibility of the test organisms to the antimicrobial effect of the plant extracts was determined using the agar well diffusion method. Standard antibiotics were used as positive control. Various culture media were used for this experiment and the minimum inhibitory concentration (MIC) was determined using the agar dilution method. Results: Extracts of D. indica fruit, leaves and stembark as well as S. mombin leaf and stembark showed significant antimicrobial activity. Activity was dose dependent and the methanol extracts had better and wider activity than the dichloromethane extracts. Spondias mombin bark had the greatest activity against most of the test organisms while the positive control drugs had better activity than the extracts. Conclusion: The leaves and stembark of Spondias mombin and the leaves, fruit and stembark of Dillenia indica have antimicrobial properties thus justifying their ethnopharmacological use in prevention and treatment of infections.

Isolation and biological evaluation of some secondary metabolites from seeds of Silybum marianum

Background: Despite the widespread ethnopharmacological use and significance of Silybum marianum (SM) seeds, most of its phytochemical and biological properties are still yet to be confirmed using validated scientific methods. Objectives: The current study was designed to investigate the phytochemical and biological properties of SM seed extract. Materials and Methods: Methanolic extract of SM (MESM) dried seeds was fractionated by column chromatography, and fractions (SA1–SA10) were evaluated for antioxidant, antimicrobial, and cytotoxic activities (brine shrimp lethality assay and antileishmanial assay). Results: All fractions showed considerable level of antioxidant potential. Free radical scavenging activity of fraction (SA9) was maximum at 80.7%. Fraction SA4 exhibited substantial total antioxidant capacity (101.81 μg/mg). In ferric-based reducing antioxidant power assay, fraction SA4 showed the highest antioxidant power (258.93 μg/mg). Phytochemical screening of the fractions (SA1–SA10) inferred that total phenolic contents were maximum in fraction SA7 (85.13 μg/mg) and total flavonoid contents were found to be highest in fraction SA1 (58.24 μgQE/mg). However, mild antibacterial and antifungal activities were shown by different fractions. To evaluate cytotoxic potential, brine shrimp lethality bioassay was performed. Among all the fractions, the fraction SA9 revealed the lowest LD50of 49.99 μg/mL, whereas all the other fractions tested demonstrated significant cytotoxic property. The results of antileishmanial assay showed that the fraction SA6 possesses the highest mortality percent (84%) compared to the other fractions. Conclusion: These findings revealed that MESM can be an important source of natural antileishmanial herb that can be used as a therapeutic alternative for leishmaniasis.

Polysaccharide composition of an anticoagulant fraction from the aqueous extract of Marsypianthes chamaedrys (Lamiaceae)

Marsypianthes chamaedrys (Lamiaceae) is a medicinal plant popularly used against envenomation by snakebite. Pharmacological studies have shown that extracts of M. chamaedrys have antiophidic, anti-inflammatory and anticoagulant properties, supporting the ethnopharmacological use. In this study, an aqueous extract of aerial parts of M. chamaedrys showed anticoagulant activity in the activated partial thromboplastin time assay (0.54 IU/mg). The bioassay-guided fractionation using ethanol precipitation and gel filtration chromatography on Sephadex G-50 and Sephadex G-25 resulted in a water-soluble fraction with increased anticoagulant activity (Fraction F2−A; 2.94 IU/mg). A positive correlation was found between the amount of uronic acids and the anticoagulant potential of the active samples. Chemical and spectroscopic analyses indicated that F2−A contained homogalacturonan, type I rhamnogalacturonan, type II arabinogalactan and α-glucan. UV and FT–IR spectra indicated the possible presence of ferulic acid. Pectic polysaccharides and type II arabinogalactans may be contributing to the anticoagulant activity of the aqueous extract of M. chamaedrys in the APTT assay.

Polyphenolic Composition and Hypotensive Effects of Parastrephia quadrangularis (Meyen) Cabrera in Rat.

Parastrephia quadrangularis (Pq), commonly called “Tola”, is widely used in folk medicine in the Andes, including for altitude sickness. In this study, polyphenolic composition was determined, and hypotensive effects were measured; the ethnopharmacological use as hypotensive was related to the presence of phenolic compounds. For this purpose, male Sprague-Dawley rats (6 to 8 weeks of age, 160 to 190 g) were fed Pq extract (10 to 40 mg/kg) for 10 days through gavage. Blood pressures and heart rate were significantly (p < 0.01) reduced in normotensive rats receiving Pq extract (40 mg/kg body weight). Pq extract induced a negative inotropic effect, and endothelium-dependent vasodilation mediated by nitric oxide (NO). Furthermore, preincubation with Pq extract significantly decreased the cytosolic calcium on vascular smooth muscle cells A7r5 in response to L-phenylephrine (PE). Seven metabolites were isolated from the Pq extract, but three flavonoids (10−4 M) showed similar vasodilation to the extract in intact rat aorta as follows: 5,3′,4′-trihydroxy-7-methoxyflavanone (2); 3,5,4′-trihydroxy-7,8,3′-trimethoxyflavone (6); and 5,4′-dihydroxy-3,7,8,3′-tetramethoxyflavone (7). The Pq extract and compounds 2 and 7 significantly (p < 0.05) reduced the contraction to Bay K8644 (10 nM, an agonist of CaV1.2 channels). Administration of Pq decreased cardiac contractility and increased endothelium-dependent and -independent vasodilation.

Phytochemical analysis, nutritional profile and immunostimulatory activity of aqueous extract from Malpighia emarginata DC leaves

Abstract The research of new natural compounds known as biomolecules increasingly demonstrates the importance of the correlation between ethnopharmacological use and the scientific evidence of its therapeutic efficacy. In this study, we investigated the main structural constituents of the plant and analyzed the phytochemical profile, antioxidant activity, antimicrobial and immunostimulatory effects on Balb/c mice splenocytes promoted by the aqueous extract obtained from leaves of Malpighia emarginata DC. The nutritional analysis of the vitamins present in the aqueous extract was performed by high performance liquid chromatography (HPLC) and colorimetric methods were used for the quantification of C vitamin, elimination of ABTS radicals, nitric oxide radicals and lipid peroxidation assays. The antibiofilm assay was performed by atomic absorbance spectrophotometry, using clinically important species of Candida spp. The measurement of cytokine levels was performed by CBA technique in flow cytometry. Results showed a high presence of glucose (0.4 g/L), lignin (20%) and pectin (21%) in organic material, as well as the prevalence of Fe2+ (103.10 μg/100 g) and Mn (10.33 μg/100 g). The aqueous extract showed the presence of C and B3 Vitamins and induced a reduction of biofilm production of C. kurei and C. parapsilosis. The immunological assay demonstrated that the extract promotes an immunostimulatory and anti-inflammatory profile, with significant production of Th2 type cytokines (IL-4, IL-10 and IL-6) and a significant reduction of nitric oxide release. The aqueous extract of leaves from M. emarginata has biological potential to be used in several diseases, as well as for future pharmacological and nutritional studies.

Polysaccharides derived from Morinda citrifolia Linn reduce inflammatory markers during experimental colitis.

There are many reports of pharmacological activities of extracts and fractions of different vegetable-derived products in the scientific literature and in folk medicine. Ethnopharmacological use of these products by various communities continues to be extensively explored, and they account for more than half of all medications used worldwide. Polysaccharides (PLS) extracted from plants such as Morinda Citrifolia Linn present therapeutic potential in treatment of inflammatory bowel diseases (IBD) such as ulcerative colitis (UC). To evaluate the anti-inflammatory action of Noni-PLS against the intestinal damage in UC induced by acetic acid in mice. In acetic acid-induced colitis, the mice were treated intraperitoneally (ip) with Noni-PLS (0.1, 0.3, and 3.0 mg/kg) or subcutaneously (sc) with dexamethasone (2.0 mg/kg) 30 min before euthanasia to determine the best dose of Noni-PLS with an anti-inflammatory effect in the course of UC. The colonic tissue samples were collected for macroscopic, wet weight, microscopic and biochemical (myeloperoxidase (MPO), glutathione (GSH), malondialdehyde (MDA), nitrate/nitrite (NO3/NO2), cytokines, cyclooxygenase (COX-2) and inducible nitric oxide (iNOS)) analyses. Treatment with Noni-PLS reduced the intestinal damage induced by acetic acid as it reduced macroscopic and microscopic scores and the wet weight of the colon. In addition, MPO activity and levels of GSH, MDA, NO3/NO2, pro-inflammatory cytokines, and COX-2 expression reduced. This study suggests that Noni-PLS exhibits anti-inflammatory action against intestinal damage by reducing inflammatory cell infiltration, oxidative stress, pro-inflammatory action of cytokines, COX-2 and iNOS expression in the inflamed colon. Noni-PLS shows therapeutic potential against inflammatory disorders like UC.

Spray Drying of a Subcritical Extract Using Marrubium vulgare as a Method of Choice for Obtaining High Quality Powder.

White horehound (Marrubium vulgare L.), is a grey-leaved perennial herb, belonging to Lamiaceae family, distributed in Eurasia and northern Africa. Despite the fact that M. vulgare has been used since ancient times in treating diverse diseases, it is only in the last decade or so that scientists have been able to lay the foundation for its potential pharmacological actions from the results observed through the prism of ethnopharmacological use of this species. The novelty of this study was that subcritical water extraction, acknowledged as a powerful extraction technology to recover phenolic compounds, was coupled with spray drying. The subcritical horehound extract, obtained using optimal process parameters, was used as a liquid feed in spray drying. Maltodextrin was used as a carrier in a concentration of 10%. Thus, two M. vulgare powders, carrier-free and 10% MD, were produced. Comprehensive powders characterization was conducted in order to evaluate their quality. Results confirmed that spray drying can be used as a method of choice for obtaining high quality horehound powders which kept the amorphous structure constant after 6 months.

The effects of Vitis vinifera L. phenolic compounds on a blood-brain barrier culture model: Expression of leptin receptors and protection against cytokine-induced damage.

The medicinal properties of grapes (Vitis vinifera L.) are well known since ancient times. Ethnobotanical grape preparations, like the Ayurvedic Darakchasava are used as cardiotonic and for the treatment of cardiovascular diseases. Dried grape products are also applied in Iranian traditional medicine for memory problems, which are linked to the pathology of brain microvessels, a special part of the cardiovascular system. The anti-inflammatory and protective effects of these traditional preparations on the cardiovascular system are related to their bioactive phenolic compounds. The blood-brain barrier (BBB), formed by brain capillaries, is not only involved in inflammatory and other diseases of the central nervous system, but also in many systemic diseases with an inflammatory component. Dietary obesity is a systemic chronic inflammatory condition in which the peripheral and central vascular system is affected. Among the cerebrovascular changes in obesity defective leptin transport across the BBB related to central leptin resistance is observed. Our aim was to study the protective effects of grape phenolic compounds epicatechin (EC), gallic acid (GA) and resveratrol (RSV) and grape-seed proanthocyanidin-rich extract (GSPE) on a cytokine-induced vascular endothelial inflammation model. Using a culture model of the BBB we investigated cytokine-induced endothelial damage and changes in the expression of leptin receptors and leptin transfer. For the BBB model, primary cultures of rat brain endothelial cells, glial cells and pericytes were used in co-culture. Cells were treated by tumor necrosis factor-α (TNF-α) and interleukin-1 β (IL-1β) (10 ng/ml each) to induce damage. Cell toxicity was evaluated by the measurement of impedance. The expression of leptin receptors was assessed by RT-qPCR and western blot. The production of reactive oxygen species (ROS) and nitric oxide (NO) were detected by fluorescent probes. GSPE (10 μg/ml), EC (10 μM), GA (1 μM) or RSV (10 μM) did not change the viability of brain endothelial cells. The gene expression of the short leptin receptor isoform, Ob-Ra, was up-regulated by GSPE, EC and RSV, while the mRNA levels of Lrp2 and clusterin, clu/ApoJ were not affected. The tested compounds did not change the expression of the long leptin receptor isoform, Ob-Rb. RSV protected against the cytokine-induced increase in albumin permeability of the BBB model. GSPE and EC exerted an antioxidant effect and GSPE increased NO both alone and in the presence of cytokines. The cytokine-induced nuclear translocation of transcription factor NF-κB was blocked by GSPE, GA and RSV. Cytokines increased the mRNA expression of Lrp2 which was inhibited by EC. RSV increased Ob-Ra and Clu in the presence of cytokines. Cytokines elevated leptin transfer across the BBB model, which was not modified by GSPE or RSV. Our results obtained on cell culture models confirm that natural grape compounds protect vascular endothelial cells against inflammatory damage in accordance with the ethnopharmacological use of grape preparations in cardiovascular diseases. Furthermore, grape compounds and GSPE, by exerting a beneficial effect on the BBB, may also be considered in the treatment of obesity after validation in clinical trials.

Eugenia pyriformis Cambess: a species of the Myrtaceae family with bioactive essential oil

Eugenia species have been appreciated for their edible fruits and medicinal properties. This paper aims to investigate the chemical composition and in vitro antileishmanial, antifungal and antiproliferative activities of essential oil from aerial parts of Eugenia pyriformis (EP-EO). The oil showed strong antileishmanial activity against promastigote forms of Leishmania amazonensis (IC50 = 2.16 µg/mL). It also exhibited high antifungal activity against Malassezia furfur (MIC = 30 µg/mL), which was determined by the broth microdilution method. Its antiproliferative activity was evaluated against the following cells: GM07429A (normal cell), MCF-7 (human breast adenocarcinoma), HeLa (human cervical adenocarcinoma) and M059J (human glioblastoma). Its major constituents, which were determined by GC-FID and GC-MS, were limonene (14.8%), nerolidol (11.0%), α-cadinol (10.3%), caryophyllene oxide (9.9%) and β-pinene (7.1%). These results showed, for the first time, the effectiveness of EP-EO as a natural product which has promising biological activities, a fact that enables its ethnopharmacological use.

Evaluation of Antipyretic and Anti-inflammatory Activity of Methonolic Extract of Andrographis paniculata in Albino Rats

The study was conducted to determine the antipyretic and anti-inflammatory activity of the methanolic extract of Andographis paniculata (APE) in albino rats. Acute oral LD50 of APE in female rats was more than 2000 mg/kg. The antipyretic activity was studied by inducing pyrexia with Brewer's yeast. A total number of thirty albino rats (200 g) were used for the study of anti-pyretic activity they were divided into five groups of six rats in each group. Group I served as control Normal saline and Group II were given brewer's yeast alone (20 ml/kg), Group III was administered standard drug Aspirin @ 100 mg/kg body wt. while groups IV and V were treated with 200 mg/kg and 400 mg/kg of Andrographis paniculata extract respectively. Pyrexia was induced by subcutaneously injecting 20% w/v brewer's suspension in below the nape of the neck of the animals. The anti-inflammatory activity of APE was assessed by measuring the reduction in carrageenan-induced paw oedema in rats. A twenty four albino rats (200 g) were used for the study of anti-inflammatory activity. Four groups were divided with six rats in each group. Group I served as control Normal saline solution and Group II was administered standard drug phenylbutazone @ 100 mg/kg While, groups III and IV were treated with 200mg/kg and 400mg/kg of APE respectively. APE (@ 400 mg/kg had significant antipyretic and anti-inflammatory activity against reduced brewer's yeast induced pyrexia and carrageenan-induced rat paw edema in rats suggesting potent antipyretic effect of APE. From these results it may be concluded that crude methanolic extract of Andrographis paniculata have significant antipyretic activity and anti-inflammatory activity that might be due to combined effect of active constituents present in plant extract this strongly support the ethno pharmacological use of the plant for the management of fever and inflammation.

Screening for bioactive secondary metabolites in Sri Lankan medicinal plants by microfractionation and targeted isolation of antimicrobial flavonoids from Derris scandens

Sri Lanka is known to have very diverse flora. Many of these species are used for plant-based remedies, which form the integral part of two Sri Lankan systems of traditional medicine, Ayurveda and Deshiya Chikitsa. Despite their widespread use, only a limited number of studies have probed into the scientific evidence for bioactivity of these medicinal plants. Such studies rarely progress to the identification of bioactive natural products. The primary aim was to develop a bioactivity screening method and apply it to 50 Sri Lankan medicinal plants where antimicrobial properties could be relevant for its traditional use. The subsequent aim was the progression into defining and characterising potent isolates within targeted compound classes from such plants, i.e. Derris scandens and its antimicrobial flavonoids. The plant collection comprised 24 species of Fabaceae, 15 Rubiaceae, 7 Solanaceae and 4 Cucurbitaceae plants. These 50 species were collected based on their ethnopharmacological importance and use in Sri Lankan traditional medicine. Crude extracts from each species were initially subjected to radial disc diffusion and microdilution assays. Subsequently, aqueous extracts of all plants were microfractionated in deep well plates using reversed-phase HPLC. Fractions were tested for antibacterial and cytotoxic activities and masses of target bioactive compounds were identified using mass spectrometry. Bioactive compounds with the masses identified through microfractions were isolated from Derris scandens using reversed-phase HPLC. The isolated pure compounds were characterised using LC-MS and NMR. Crude aqueous extracts from 19 species showed activity against Gram-positive bacteria (Staphylococcus aureus and Bacillus cereus) in the radial disc diffusion assay. Crude aqueous extracts from 34 plant species and organic extracts from 46 plant species were active against S. aureus (≤4 mg mL-1) in the microdilution assay. Microfractionation demonstrated antibacterial activity for 19 plants and cytotoxicity for 6 plants. Furthermore, target bioactive compounds and their molecular ions were identified during microfractionation. Dalpanitin and vicenin-3, two of the flavonoids isolated from Derris scandens gave MICs of 23 μg mL-1 against S. aureus. Dalpanitin also exhibited relevant MICs on Gram-negative bacteria (94 μg mL-1 against Escherichia coli and Pseudomonas aeruginosa). The microfractionation protocol developed in this study enabled time-efficient screening of many plants species, using a small quantity of sample material. In addition, microfractionation served as a guiding tool for identifying individual antimicrobial compounds. Through this process, flavonoids were isolated from Derris scandens, out of which dalpanitin and vicenin-3 showed activity in the low micromolar range. The high hit rate for in vitro antibacterial properties from this ethnopharmacologically guided sample collection gives credence to Sri Lankan traditional herbal medicine as a source for drug discovery.

In vitro inhibition of HIV-1 reverse transcriptase and anti-inflammatory activities of some herbal concoctions sold in the Limpopo Province

HIV-1 is the principal virus that causes acquired immunodeficiency syndrome (AIDS), a condition characterized by extreme immunosuppression and inflammation that leads to life-threatening opportunistic infections. The pitfalls of current anti-retroviral therapies are the lack of medical access in developing countries, adverse side effects and the rise of resistant strains. Natural products used in traditional medicine that can interfere with the replication of the HIV serve as good sources of anti-HIV drugs and anti-inflammation agents. The aim of this study was to validate the anti-HIV and anti-inflammatory properties of some herbal concoctions sold in the Limpopo Province, reported to treat HIV and inflammation related symptoms. The anti-HIV activity was evaluated using the HIV-1 reverse transcriptase (RT) inhibitory bioassay. Anti-inflammatory activity was determined using cyclooxygenase 1 and 2 inhibitory assays. Compared to the controls, Lamivudine, Zidovudine, Lopinavir and Ritonavir, the laboratory concoction showed the highest HIV-reverse transcriptase inhibition (IC50: 2.90 μg/mL), followed by herbal concoction 5 (IC50: 11.74 μg/mL). It is likely the herbal concoctions induced a conformational change on the HIV-RT by binding to its allosteric sites, thus inactivating it. Generally, the herbal concoctions achieved higher percentage inhibitions (≥70%) for COX-2 than for COX-1. The anti-inflammatory activity of the herbal concoctions thus may also assist to alleviate pain in the patients. This research validated the ethnopharmacological use of herbal concoctions towards the treatment of HIV infections. However, more studies are needed to further elucidate the pharmacokinetics and toxicological effects of these medicines.