Aqueous Methanol Fractions Research Articles

Biological activity of phenolics enriched extracts from industrial apple pomace

Apple processing industries around the globe generate huge quantity of pomace as waste material, whose sustainable and cost-effective utilization continually being attempted. In general, industrial apple pomace is a conglomeration of different varieties, however, rich in array of inherited nutritional and phytochemical constituents. In present study, aqueous-ethanol, -methanol and -acetone mediated extractions of phenolics from industrial apple pomace was performed. The obtained extracts from respective solvents were further partitioned with ethyl acetate. The phytochemical evaluation of ethyl acetate fractionated aqueous acetone fraction (APA1), aqueous ethanol fraction (APE1) and aqueous methanol fraction (APM1) by high pressure liquid chromatography and electrospray-ionization quadrupole time-of-flight tandem mass spectrometry showed the presence of phloridzin, phloretin, quercitrin and quercetin as major constituents. The cytotoxic effects of APA1, APM1, and APE1 fractions were evaluated against human cancer cell lines i.e. cervical squamous cell carcinoma (SiHa), oral carcinoma (KB) and colorectal adenocarcinoma (HT-29) cell lines. Significant differences were obtained for oral cancer cells in time and dose dependent manner. Cytotoxicity data validated by clonogenic cell survival assay showed significant inhibition of KB cancer cells by APA1 and APE1 after 24 and 36 h. Caspase 3/7 activity of fractions was also found to be increased in time dependent manner. Furthermore, the flow cytometer analysis revealed that KB cells follow apoptotic and necrotic mechanism for cell death after treatment with APA1 and APE1 fractions for 24 and 30 h. Results also showed significant nitric oxide inhibition in lipopolysaccharide– interferon gamma stimulated mice peritoneal macrophages at different concentrations. Present study established that industrial apple pomace indeed has substantial amount of bioactive phenolics that can be bioprocessed to develop variety of health supplements.

In vitro antioxidant, anti‐inflammatory and antihypertensive activities of methanol leaf extract of Peasea americana

Hypertension is a cardiovascular disease of remarkable health and economic importance. Management of hypertension with orthodox medicine is associated with side effects and is unaffordable to a larger percentage of people in Africa. P. americana leaf extract (PALE) is commonly used in the management of hypertension in Nigeria trado‐medicine. In this study, we investigated the antioxidant, anti‐inflammatory and anti‐hypertensive activities of the crude methanol extract, fractions and phytochemicals of P. americana leaf. The in vitro‐antioxidant assays were carried out to evaluate the free radicals scavenging activities of PALE on 1, 1‐diphenyl‐2‐picrylhydroxyl (DPPH), 2, 2‐azenobis‐3‐ethyl benzothiozoline‐6‐sulfonic acid (ABTS), hydroxyl and nitric oxide radicals. In addition, metal chelating and ferric reducing power assay were done. Anti‐inflammatory activities were evaluated using 15‐1ypoxygenase enzyme inhibition assay. Absorption were read spectophotometrically and 1C50 was calculated using a standard calibration curve. Phytocompound isolation was by extract fractionation, column, thin layer and preparatory thin layer chromatography. Crude methanol extracts, N‐hexane, ethyl acetate, chloroform, N‐butanol and aqueous methanol fractions had significantly higher anti‐oxidant activities when compared with gallic acid. There anti‐inflammatory activity is comparable to that of Indomethacin and antihypertensive activity for PALE (with IC50 of 135.99±10.06) is significantly lower when compared with captopril (with IC50 41.86±2.10). Four pure phytochemicals were isolated and named LEACA, LEACB, LEACC, and LEACD. The isolated compounds had significantly higher antioxidant activities on DPPH, ABTS, and hydroxyl and nitric oxide radicals when compared with gallic acid. The anti‐hypertensive activities of the isolated phytocompounds from the ethyl acetate fraction of the leaf were significantly higher (with IC50 of 24.83±0.63, 9.48±1.45, 2.79±1.35 and 1.59±0.48 respectively) than that of captopril (with 1C50 of 41.89±2.10). The result obtained from this study suggest that P. americana leaf has free radical scavenging and antioxidant activities, anti‐inflammatory and anti‐hypertensive activities and can ameliorate oxidative stress, and inflammation associated with hypertension.

Antimalarial Activity of Meriandra dianthera Leaf Extracts in Plasmodium berghei-Infected Mice.

Objective To evaluate the antimalarial effect of aqueous methanolic extract and solvent fractions of Meriandra dianthera leaves against Plasmodium berghei in mice model. Method M. dianthera leaves were extracted with 80% methanol and dried. The dried crude extract was then defatted and further fractionated with chloroform, ethyl acetate, and butanol. Acute oral toxicity test was performed as per the Organization for Economic Cooperation and Development guideline 425. Peter's 4-day suppressive test was used to determine the in vivo antimalarial activity of the extract and fractions. Result The crude leaf extract of Meriandra dianthera leaves against P < 0.001) chemosuppression compared to the negative control. Both the extract and fractions were able to prevent P. berghei induced body weight loss and body temperature reduction and also increased the survival time of the mice as compared to the negative control. The aqueous methanolic leaf extract of M. dianthera leaves were extracted with 80% methanol and dried. The dried crude extract was then defatted and further fractionated with chloroform, ethyl acetate, and butanol. Acute oral toxicity test was performed as per the Organization for Economic Cooperation and Development guideline 425. Peter's 4-day suppressive test was used to determine the Conclusion The extracts of M. dianthera leaves showed promising antimalarial activity, with no sign of toxicity and therefore may support its traditional use for the treatment of malaria.M. dianthera leaves were extracted with 80% methanol and dried. The dried crude extract was then defatted and further fractionated with chloroform, ethyl acetate, and butanol. Acute oral toxicity test was performed as per the Organization for Economic Cooperation and Development guideline 425. Peter's 4-day suppressive test was used to determine the

Gastroprotective Effects of β-sitosterol and β-sitosterol-3-O-β-D-glucopyranoside from Bridelia ferruginea Stem Bark

The methanol extract of Bridelia ferruginea Benth. (Euphorbiaceae) stem bark (BFME) was partitioned in chloroform-methanol-water (2:2:1) mixture to obtain the Chloroform (CF) and Aqueous Methanol (AMF) fractions. The BFME, CF and AMF were screened for antiulcer activity using indomethacin-induced ulcer as activity guide. The CF provided the highest gastroprotection and was subsequently fractionated in a silica gel (60-200 mesh) column eluted with different mixtures of n-hexane and ethyl acetate (100:0; 95:5; 90:10; 80:20) to obtain six fractions (I-VI). Fractions III and VI offered the highest protection against indomethacin-induced ulcer and were further purified in a sephadex LH-20 column eluted with methanol to yield two compounds, BF1 and BF2. Using Nuclear Magnetic Resonance (1H-NMR, 13C-NMR) and electron impact mass spectroscopies, BF1 and BF2 were confirmed to be β-sitosterol and β-sitosterol-3-O-βD glucopyranoside respectively. The BFME, fractions, β-sitosterol and β-sitosterol-3-O-β-D-glucopyranoside elicited dose-related and significant (P<0.05) protection against various ulcers in rats. β-sitosterol, 100 and 300 mg/kg, produced 79.70, 82.18, 42.31, 44.87, 65.97, 70.83, 80.22 and 87.91% gastroprotection; while β-sitosterol-3-O-β-D-glucopyranoside, 100 and 300 mg/kg, caused 69.80 and 74.26, 33.33, 35.26, 84.03, 95.83, 83.52 and 85.71% gastroprotection against indomethacin-, ethanol-, cold restraint stress- and pylorus ligation- induced ulcers, respectively. Results demonstrated gastroprotective effects of B. ferruginea stem bark, attributable to β-sitosterol and β-sitosterol-3-O-β-D-glucopyranoside.

Toxicological study of aqueous-methanol solvent fraction of methanol extract of Dacryodes edulis leaves

Dacryodes edulis (G. Don) H.J. Lam) is the most popular species under the genus Dacryodes. It is well known for its nutritional and ethno-medicinal uses in South-eastern and South-western Nigeria. This study was aimed to evaluate the toxicity of the aqueous-methanol fraction of crude methanol extract of Dacryodes edulis leaves (AMDE). The test rats were randomized to groups of single oral treatment of AMDE (10−5000 mg/kgbw) for the acute toxicity study. They were monitored for obvious signs of behavioural change and mortality. For the subacute toxicity study, the rats were randomized to three daily treatment groups (of 200, 400 and 600 mg/kgbw of AMDE) for 28 days. The fourth group (control) received 2.5 %v/v DMSO. At the end of the experiment, blood samples were collected for hematology and clinical chemistry evaluation. The histopathology of the livers and kidneys were assessed using the excised organs. The cytotoxicity and genotoxicity of AMDE were also evaluated using Allium cepa model. The result showed that acute administration of AMDE, up to a dose of 5000 mg/kgbw did not result in mortality of the test rats. The observed median lethal dose (LD50) was greater than 5000 mg/kgbw. The subacute oral administration of AMDE for 28 days showed no significant (p > 0.05) effect on liver function, kidney function indices, organ - body weight ratio, but significantly (p < 0.05) decreased erythrocytic indices: red blood cells, haematocrit, and haemoglobin at 600 mg/kgbw. The Allium cepa assay revealed a non-significant reduction in mitotic index and low chromosomal aberrations of the treated groups. In conclusion, the aqueous-methanol solvent fraction of methanol extract of Dacryodes edulis leaves, AMDE is relatively safe. However, there are strong indications that it may contain compounds that are cytotoxic and reduces erythrocytic indices including red blood cells at high doses. Thus, adequate care should be taken in dosing and administering the extract to avert anaemic condition.

Anti-adipogenic Effect of Homoisoflavonoids from Portulaca Oleracea (P06-015-19)

ObjectivesTo search for anti-adipogenic components from natural plants, this study evaluated inhibitory effects of active components from Portulaca oleracea on adipogenic differentiation. MethodsThe crude extract was partitioned into n-hexane, 85% aqueous methanol, n-butanol, and water fractions. By anti-adipogenic assay-guided isolation method, 4 kinds of homoisoflavonoids (1–4) were isolated from 85%aq. MeOH fraction. Their inhibitory effects on adipogenic differentiation were evaluated by Oil red O intracellular lipid staining, and measuring glycerol release, glucose consumption and adiopogenesis-related gene expression in 3T3-L1 cells. ResultsThe administration of homoisoflavonoids 1–4 to adipocyte cells decreased the lipid accumulation and glucose consumption, and increased the release of glycerol into culture medium. In particular, homoisoflavonoid 3 effectively down-regulated the adipogenic transcription genes such as peroxisome proliferator activated receptor-γ (PPARγ) and CCAAT/enhancer-binding proteins (C/EBPα), and adipogenic target genes such as fatty acid binding protein 4 (FABP4), fatty acid transport protein 1 (FATP1), and acyl-CoA synthase 1 (ACS1). ConclusionsThis study suggested that homoisoflavonoids 3 held notable potential to be used for the treatment and prevention of obesity and related disorders. Funding SourcesThis research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education and the Center for Women In Science, Engineering and Technology (WISET) Grant funded by the Ministry of Science and ICT (MSIT) under the Program for Returners into R&D.

Evaluation of anti-adipogenic active homoisoflavonoids from Portulaca oleracea.

This study was performed to isolate antiobesity components from the crude extract of Portulaca oleracea. The crude extract was partitioned into n-hexane, 85% aqueous methanol, n-butanol, and water fractions. Their effects on adipogenic differentiation were evaluated in 3T3-L1 cells. Among the solvent fractions from P. olearacea, the 85% aq. MeOH effectively reduced the levels of lipid accumulation. Further purification of 85% aq. MeOH led to the isolation of the known homoisoflavonoids 1-4, as the active substances. The administration of homoisoflavonoids to adipocyte cells decreased the lipid accumulation and glucose consumption and increased the release of glycerol into culture medium. In particular, homoisoflavonoid 3 effectively down-regulated the adipogenic transcription genes such as peroxisome proliferator activated receptor-γ (PPARγ) and CCAAT/enhancer-binding proteins (C/EBPα), and adipogenic target genes such as fatty acid binding protein 4 (FABP4), fatty acid transport protein 1 (FATP1), and acyl-CoA synthase 1 (ACS1).

Phytochemical screening and antibacterial activity of Aspilia africana on some gastrointestinal tract pathogens

The main aim of this study was to determine the phytochemical constituents and antibacterial activities of the leaf extracts of A. africana. The powdered leaf of A. africana was extracted using 70% methanol and partitioned into n-hexane, chloroform, ethyl acetate and aqueous methanol fractions. The extract and fractions were phytochemically screened and agar well dilution technique was used to evaluate the antibacterial activity against some gastrointestinal tract pathogens (Escherichia coli, Staphylococcus aureus, Shigella dysentriae, Salmonella typhi, Salmonella paratyphi A, Salmonella paratyphi B and Salmonella paratyphi C). The phytochemical analysis of A. africana showed the presence of alkaloids, terpenoids, tannins, flavonoids, anthraquinones and saponins in the methanol extract but varied across the fractions. The methanol extract and fractions inhibited the growth of gastrointestinal tract pathogens but their effectiveness varied with the concentrations. The methanol crude extract showed the best antibacterial activity with zone of inhibition ranging from 15-18 mm followed by aqueous soluble methanol fraction (10-15 mm) and ethylacetaate fraction (9-13 mm) respectively. The n-hexane and chloroform fraction showed no activity. Generally, the diameter zone of inhibition increased with increased extract concentrations. The antibacterial activity of the various extracts of the leaf was comparable to the reference antibiotics, though the antibiotics showed better inhibitory property (p<0.05) in some cases on the test isolates than the leaf extracts. The study showed that the leaves of A. africana possessed inhibitory properties due to the detected phytochemicals, thus validates the traditional uses of the plant for the treatment of gastrointestinal tract infections.

In vitro and in vivo anti-diabetic and anti-oxidant activities of methanolic leaf extracts of Ocimum canum.

Background:Diabetes is a metabolic disorder with a highly complex, multifaceted and intricate etiologies and thus may require management options that proffers multimodal mechanism of action. This present study evaluated the antidiabetic and antioxidant potential of the methanolic extract/fractions of leaves of Ocimum canum.Methods:The antidiabetic potential was evaluated and using STZ-induced diabetic Wistar rat model (in vivo) and inhibition of α-amylase and α-glucosidase activity (in vitro). Antioxidant activity was assessed in vitro by free radical scavenging and reducing power assays and in vivo via monitoring SOD and CAT activities; GSH and MDA levels.Results:The total phenolic content (221.0±3.0 mg catechol/g of sample) and tannins (146.0±4.0 mg tannic acid/g of sample) of the crude extract; and flavonoid of the aqueous-methanol fraction (216.0.0±1.0 mg of rutin/g of sample) were found to be significantly higher relative to others. The crude extract and the aqueous-methanol fraction exhibited a significantly (p<0.05) higher percentage reduction in fasting blood glucose and a concomitant increase in serum insulin level relative to the diabetic control group. The highest radical scavenging activity and reducing power were observed in the aqueous-methanol fraction. The aqueous-methanol solvent fraction also significantly reversed the alterations in oxidative stress markers occasioned by the diabetic condition.Conclusion:In conclusion, the result of the present study has demonstrated evidently that extracts of Ocimum canum leaves ameliorates hyperglycemia and the associated oxidative stress in STZ-induced rats.

Methanol-Extract/Fractions of Dacryodes edulis Leaves Ameliorate Hyperglycemia and Associated Oxidative Stress in Streptozotocin-Induced Diabetic Wistar Rats.

The present study evaluated the antidiabetic and antioxidant potential of the methanolic extract/solvent fractions of the leaves of Dacryodes edulis using a streptozotocin (STZ)-induced diabetic Albino Wistar rat model. The fasting blood glucose/insulin levels and inhibition of α-amylase and α-glucosidase were determined. Antioxidant activity was assessed in vitro by 2,2-diphenyl-1-picrylhydrazyl, hydroxyl, superoxide scavenging, reducing power, and total antioxidant capacity assays and in vivo by monitoring catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) activities and reduced glutathione (GSH) and malondialdehyde (MDA) levels. The aqueous-methanol fraction exhibited the highest and significant (P < .05) reduction in fasting blood glucose (FBG; 54.03%) with a concomitant inhibition of α-amylase and α-glucosidase activities. The ethyl acetate fraction also exhibited a significant (P < .05) reduction in FBG and an increase in insulin levels in the treated diabetic Wistar rats. A significantly (P < .05) higher reducing power and radical scavenging activity was observed in the aqueous-methanol and ethyl acetate fractions. The aqueous-methanol and ethyl acetate fractions also significantly (P < .05) reversed the alterations in oxidative stress markers (GSH, MDA, CAT, and SOD) observed in the diabetic control group. In conclusion, the study demonstrated that the methanol extract of Dacryodes edulis ameliorates hyperglycemia and the associated oxidative stress in STZ-induced diabetic Wistar rats. These observed activities are largely due to the compounds that partitions into the aqueous-methanol (55:45) solvent fraction. This provides scientific evidence for the use of this plant extract in folk medicine and also a baseline data for its further characterization. Further work should be carried out to characterize the aqueous-methanol solvent fractions for the active compounds.

Extraction and Fractionation of Whole Black Seed Plantain (Plantago rugelii Decne) for in-vitro Antioxidant, Antibacterial and Phytochemical Screening

This study was designed to explore new antioxidant and antimicrobial agents from the methanol whole plant crude extract and fractions of Plantago rugelii . The methanol extract and its fractions were prepared and screened for its phytochemical composition, in-vitro antioxidant potential and challenged with common pathogenic microorganisms for its antimicrobial activities using standard procedures. The phytochemical analysis revealed the presence of various pharmaceutically active secondary metabolites like alkaloids, phenolic, flavonoids, carbohydrates, glycosides, sterols, etc. In the DPPH assay, the aqueous methanol fraction was found to be the most effective among all the fractions in comparison to the ascorbic acid standard. Using the zone of inhibition as inhibitory parameter, the crude methanol extract exhibited the best antibacterial activity when challenged against all the clinical isolates except Klebsiella pneumoniae . The aqueous methanol extract exhibited the best percentage fungal inhibition when compared to other fractions. The standard drugs ciprofloxacin and fluconazole exhibited a near 100% activity except with Proteus vulgaris where the crude methanol extract has a higher value.This research holds promise for the exploration of various potentially active secondary metabolites which would help in developing pharmaceuticals, especially antioxidant and antimicrobial drugs. The isolation and characterization of the exact metabolites responsible for these activities is therefore recommended.

Effect of extracts from Stachys sieboldii Miq. on cellular reactive oxygen species and glutathione production and genomic DNA oxidation

Objective: To evaluate the antioxidant activity of extracts and fractions from Stachys sieboldii Miq., and to examine its effect on the cellular reactive oxygen species (ROS) and glutathione (GSH) production and genomic DNA oxidation in HT-1080 cells. Methods: The ROS generation induced by H2O2 was measured by the dichlorofluorescein-diacetate assay. GSH levels were measured using a fluorescent method with mBBr. Genomic DNA oxidative damage was measured with levels of oxidative DNA induced by the reaction of ferritin with H2O2. Results: The n-hexane, 85% aqueous methanol and n-butanol fractions (0.05 mg/mL concentrations) inhibited H2O2-induced ROS generation by 63%, 35% and 45%, respectively. GSH levels were significantly increased in both acetone+methylene chloride and methanol extracts (P

&lt;i&gt;In vivo&lt;/i&gt; antitrypanosomal effects of stem-bark extracts of &lt;i&gt;Securidaca longipedunculata&lt;/i&gt; in rats experimentally infected with &lt;i&gt;Trypanosoma brucei brucei&lt;/i&gt;

The efficacy of stem-bark extracts of Securidaca longipedunculata against Trypanosoma brucei brucei infected rats was investigated. For curative study, forty adult Wistar rats of both sexes were randomly divided into 8 groups of 5 rats each. Each rat was infected with 10 6 cells of trypanosomes per ml of blood intraperitoneally (ip). Rats in groups 1 and 2 received the crude methanol extract (CME) at 0.7 and 0.35 mg/kg, respectively. Similarly, rats in groups 3 and 4 received ethyl acetate fraction (EAF) at 0.7 and 0.35 mg/kg, respectively; while 5 and 6 were treated with 0.9 and 0.45 mg/kg of aqueous methanol fraction (AMF), respectively. Rats in groups 7 and 8 were treated with diminazene aceturate (3.5 mg/kg) and phosphate buffered saline, PBS (2 ml/kg), respectively. Four rats (group 9) were neither infected nor treated and served as neutral control. In the prophylactic studies, 25 rats of both sexes were randomly divided into V groups of 5 rats each. Rats in groups I, II, and III were pre-treated with CME at 0.7 mg/kg i.p. for 3, 5 and 7 days, respectively; while group IV received PBS for 7 days and served as negative control. The rats were then individually infected with 10 6 parasites per ml of blood on days 3, 5 and 7 for groups I, II and III, respectively. Rats in group V were neither treated nor infected and served as neutral control. CME of S. longipedunculata suppressed level of parasitaemia and prolonged the survival period of rats when compared to other groups (P<0.05). Pre-treatment of animals with CME before challenge with the parasite could not prevent infection. Thus, stem-bark extract of S. longipedunculata exhibited some levels of curative antitrypanosomal effect against T. brucei brucei infection in rats despite its low margin of safety. Keywords : Curative effect, High toxicity, In vivo, Phytochemical screening, Prophylactic effect

Anti-arthritic activity of aqueous-methanolic extract and various fractions of Berberis orthobotrys Bien ex Aitch

BackgroundThe roots and stem bark of Berberis orthobotrys (Berberidaceae) have long been used traditionally to treat joint pain. Though, it has not been pharmacologically assessed for rheumatoid arthritis. The current study explores anti-arthritic activity and phytochemical analysis of aqueous-methanolic extract (30:70) and fractions (ethyl acetate, n-butanol, and aqueous) of Berberis orthobotrys roots.MethodsAnti-arthritic potential was evaluated in vitro using protein denaturation (bovine serum albumin and egg albumin) and membrane stabilization methods at 12.5–800 μg/ml concentration and in vivo via turpentine oil, formaldehyde and Complete Freund Adjuvant (CFA) models at 50, 100 and 150 mg/kg doses. Also, in vitro antioxidant ability was appraised by reducing power assay. Moreover, total flavonoid content, Fourier transform infrared spectroscopy and High performance liquid chromatography of n-butanol fraction were performed.ResultsThe results revealed concentration dependent inhibition of albumin denaturation and notable RBC membrane stabilization, with maximum results obtained at 800 μg/ml. Similarly, plant exhibited dose dependent anti-arthritic effect in turpentine oil and formaldehyde models, with maximum activity observed at 150 mg/kg. The results of CFA model depicted better protection against arthritic lesions and body weight alterations. Also, B.orthobotrys remarkably ameliorated altered hematological parameters, rheumatoid factor and positively modified radiographic and histopathological changes. Additionally, plant exhibited remarkable anti-oxidant activity. Moreover, phytochemical analysis revealed polyphenols and flavonoids.ConclusionTaken together, these results support traditional use of B.orthobotrys as potent anti-arthritic agent that may be proposed for rheumatoid arthritis treatment.

여정실의 항산화 활성

The objective of this study is to evaluate the antioxidant activity of the fruits of Ligustrum japonicum. The crude extract was successively fractionated into n-hexane, 85% aqueous methanol (85% aq.MeOH), n-butanol (n-BuOH), and water fractions by means of solvent polarity. The crude extract and its solvent fractions were evaluated for their antioxidant effect by four different assay systems: scavenging power on peroxynitrite and intralcellular ROS produced in HT-1080 cells; DNA oxidation inhibition; ferric reducing antioxidant power (FRAP). The n-BuOH fraction exhibiting potent antioxidant activity was further purified by C18 silica gel column chromatography and RP-HPLC to give tyrosol (1) and salidroside (2). The structure of isolated compounds was determined by extensive 2 D NMR experiments such as <TEX>$^1H$</TEX> COSY, NOESY, HSQC and HMBC as well as by comparison with the published spectral data.

The Indian Magical Herb 'Sanjeevni' (Selaginella bryopteris L.) - A Promising Anti-inflammatory Phytomedicine for the Treatment of Patients with Inflammatory Skin Diseases.

ObjectivesSelaginella bryopteris L. (family: Selaginaceae), is often used in traditional Indian systems of medicine for the prevention and cure of several disorders and for the treatment of patient with spermatorrhoea, venereal disease, constipation, colitis, urinary tract infections, fever, epilepsy, leucorrhoea, beri-beri and cancer. It is also used as a strength tonic. This study aimed to evaluate the mechanisms underlying the anti-inflammatory effects of topically administered aqueous, polar and non-polar methanolic fractions (10 mg/20 μL) of Selaginella bryopteris.MethodsAn acute oral toxicity study of Selaginella bryopteris at doses from 250 to 2,000 mg/kg body weight (bw) was performed. Aqueous, polar and non-polar methanolic extracts (10 mg/20 μL) applied topically for 5 days were evaluated for their anti-inflammatory effects against 12-tetra-O-decanoyl phorbol acetate (TPA)-induced inflammation by using the redness in the ear, the ear’s weight (edema), oxidative stress parameters, such as lipid-peroxide (LPO) and nitric oxide (NO), and the pro-inflammatory cytokines involved in inflammation, such as tumour necrosis factor (TNF)-α, interleukin (IL)-1β and IL-6. Indomethacine (0.5 mg/20 μL) was used for the positive control.ResultsSelaginella bryopteris produced no mortalities when administered orally at doses from 250 to 2,000 mg/kg bw. Topical treatment with the non-polar methanolic fraction (10 mg/20 αL) significantly suppressed redness (2.4 ± 0.5) and edema (30.4 ± 1) and effectively reduced the LPO level (32.3 ± 3.3). The NO level was (8.07 ± 0.55), and the TNF-α, IL-1β, and IL-6 levels were decreased to 69.6 ± 15.5, 7.7 ± 4.8 and 82.6 ± 5.9, respectively.ConclusionThis study demonstrated for the first time the mechanisms underlying the anti-inflammatory effect of medicinal plants like Selaginella bryopteris and quantified the pharmacological interactions between them. The present study showed this herbal product to be a promising anti-inflammatory phytomedicine for the treatment of patients with inflammatory skin diseases.

A controlled study to investigate anti-diarrhoeal effect of the stem-bark fractions of Terminalia avicennioides in laboratory animal models

Due to the shortcomings associated with modern synthetic antidiarrhoeal drugs, it is important to find newer, safer and cheaper antidiarrhoeal agents from natural sources. The study was conducted to evaluate the anti-diarrhoeal activity of the fractions of the stem-bark of Terminalia avicennioides in laboratory animal models. The effect of different concentrations (1.0×10−3, 2.0×10−3, 4.0×10−3 and 8.0×10−3mg/mL) of the aqueous methanol (AMF), ethyl acetate (EAF) and hexane (HXF) fractions of T. avicennioides were tested against spontaneous and acetylcholine-induced contractions of rabbit jejunum as well as on histamine-induced contraction of guinea pig ileum. Similarly, the effects of the AMF on gastro-intestinal transit time, castor oil-induced diarrhoea and castor oil-induced enteropooling were evaluated. The AMF, EAF and HXF at concentrations of 1.0×10−3, 2.0×10−3, 4.0×10−3 and 8.0×10−3mg/mL attenuated the contractile effects of both the spontaneous and acetylcholine-induced contractions of rabbit jejunum and that of histamine-induced contraction of guinea pig ileum in a concentration-dependent manner. The AMF at doses of 200, 300 and 500mg/kg produced significant (p<0.05) reductions in gastrointestinal transit time of charcoal and incidence of castor oil-induced diarrhoea in mice relative to the untreated control. Similarly, at doses of 300 and 500mg/kg, AMF significantly (p<0.05) reduced the weight and volume of intestinal fluid in the treated mice when compared to the untreated animals. The results of this study showed that the stem-bark of T. avicennioides possesses spasmolytic effect and could be a potential antidiarrhoeal agent. However, detailed pharmacological trials are required to justify the clinical use of the plant for treating diarrhoea.

Nuclear magnetic resonance based metabolomic differentiation of different Astragali Radix

Astragali Radix (AR) is one of the most popular herbal medicines in traditional Chinese medicine (TCM). Wild AR is believed to be of high quality, and substitution with cultivated AR is frequently encountered in the market. In the present study, two types of ARs (wild and cultivated) from Astragalus membranaceus (Fisch.) Bge. and A. membranaceus var. mongholicus (Bge.) Hsiao, growing in different regions of China, were analyzed by NMR profiling coupled with multivariate analysis. Results showed that both could be differentiated successfully and cultivation patterns or growing years might have greater impact on the metabolite compositions than the variety; the metabolites responsible for the separation were identified. In addition, three extraction methods were compared and the method (M1) was used for further analysis. In M1, the extraction solvent composed of water, methanol, and chloroform in the ratio of 1 : 1 : 2 was used to obtain the aqueous methanol (upper layer) and chloroform (lower layer) fractions, respectively, showing the best separation. The differential metabolites among different methods were also revealed. Moreover, the sucrose/glucose ratio could be used as a simple index to differentiate wild and cultivated AR. Meanwhile, the changes of correlation pattern among the differential metabolites of the two varieties were found. The work demonstrated that NMR-based non-targeted profiling approach, combined with multivariate statistical analysis, can be used as a powerful tool for differentiating AR of different cultivation types or growing years.

Evaluation of anticonvulsant potential of aqueous methanolic extract and various fractions of Ajuga bracteosa wall

We aim to assess the anticonvulsant potential of Ajuga bracteosa Wall. The aqueous methanol extract (30:70) of Ajuga bracteosa and its n-hexane, chloroform and ethyl acetate fractions were prepared. Anticonvulsant effect was evaluated using different animal models of epilepsy. Crude extract along with its fractions at 500 and 1000 mg/kg doses, administered orally to albino mice were appraised against pentylenetetrazole-induced, strychnine-induced and picrotoxin-induced convulsions. Moreover, effect of test substances on brain GABA level and phenobarbitone induced hypnosis was also studied at dose level of 350 and 400 mg/kg. The plant extract and its fractions produced potent dose dependent anticonvulsant activity in all models of epilepsy. In addition, plant extract and fractions increased brain GABA level and potentiated phenobarbitone-induced sleep duration. From the results it can be deduced that Ajuga bracteosa possess potent anticonvulsant activity, supporting its folkloric use in the treatment of epilepsy.

Toxicity screening of 13 Gambierdiscus strains using neuro-2a and erythrocyte lysis bioassays

Species in the epi-benthic dinoflagellate genus Gambierdiscus produce ciguatoxins (CTXs) and maitotoxins (MTXs), which are among the most potent marine toxins known. Consumption of fish contaminated with sufficient quantities of CTXs causes Ciguatera Fish Poisoning (CFP), the largest cause of non-bacterial food poisoning worldwide. Maitotoxins, which can be found in the digestive system of fish, could also contribute to CFP if such tissues are consumed. Recently, an increasing number of Gambierdiscus species have been identified; yet, little is known about the variation in toxicity among Gambierdiscus strains or species.This study is the first assessment of relative CTX- and MTX-toxicity of Gambierdiscus species from areas as widespread as the North-Eastern Atlantic Ocean, Pacific Ocean and the Mediterranean Sea. A total of 13 strains were screened: (i) seven Pacific strains of G. australes, G. balechii, G. caribaeus, G. carpenteri, G. pacificus, G. scabrosus and one strain of an undetermined species (Gambierdiscus sp. Viet Nam), (ii) five strains from the North-Eastern Atlantic Ocean (two G. australes, a single G. excentricus and two G. silvae strains), and (iii) one G. carolinianus strain from the Mediterranean Sea. Cell pellets of Gambierdiscus were extracted with methanol and the crude extracts partitioned into a CTX-containing dichloromethane fraction and a MTX-containing aqueous methanol fraction. CTX-toxicity was estimated using the neuro-2a cytoxicity assay, and MTX-toxicity via a human erythrocyte lysis assay.Different species were grouped into different ratios of CTX- and MTX-toxicity, however, the ratio was not related to the geographical origin of species (Atlantic, Mediterranean, Pacific). All strains showed MTX-toxicity, ranging from 1.5 to 86pg MTX equivalents (eq) cell−1. All but one of the strains showed relatively low CTX-toxicity ranging from 0.6 to 50 fg CTX3C eq cell−1. The exception was the highly toxic G. excentricus strain from the Canary Islands, which produced 1426 fg CTX3C eq cell−1. As was true for CTX, the highest MTX-toxicity was also found in G. excentricus. Thus, the present study confirmed that at least one species from the Atlantic Ocean demonstrates similar toxicity as the most toxic strains from the Pacific, even if the metabolites in fish have so far been shown to be more toxic in the Pacific Ocean.