EtOAc Fraction Research Articles

Achillea millefolium L. ethyl acetate fraction induces apoptosis and cell cycle arrest in human cervical cancer (HeLa) cells

This study aimed to develop a comprehensive quantification of 20 bioactive compounds in Achillea millefolium L (A. millefolium) fractions obtained by different solvents (petroleum ether (pet-ether), ethyl acetate (EtOAc), methanol (MeOH) and water) using HPLC quantitative analysis. Additionally, total flavonoids (TF), total phenolics (TP), antioxidants (DPPH, ABTS) and cytotoxic activities of A. millefolium fractions on k562, HeLa, MCF7, A431 and A549 Cell Lines were evaluated using MTT assay [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] and compared to doxorubicin as a standard. According to the obtained results, TP ranged from 3.8 to 91.85 mg gallic acid equivalent (GAE)/g fraction, while TF ranged from 9.68 to 79.50 mg catechin equivalent (CE)/g generally. The lowest IC50 values were recorded for doxorubicin compared by all fractions except for EtOAc fraction against Hela cells, followed by water fraction, which showed the highest anticancer potential against MCF-7 cells. In contrast, the pet-ether fraction showed the lowest IC50 value against K562 cells. The cell cycle profile and apoptosis analysis of the EtOAc fraction was studied in HeLa cells. EtOAc fraction caused preG1 apoptosis and cell growth arrest in G2/M in HeLa cancer cells. Overall, this study heightens the importance of A. millefolium, which may be a promising and valuable source of natural antioxidant and anticancer agents to be used in the development of new functional and therapeutic compounds in the future.

Isolation, Gastroprotective Effects and Untargeted Metabolomics Analysis of Lycium Minutifolium J. Remy (Solanaceae).

Lycium minutifolium J. Remy (Solanaceae) is commonly used as an infusion in traditional medicine to treat stomach pain, meteorism, intestinal disorders, stomach ailments, and other severe problems including prostate cancer and stomach cancer. From the EtOAc extract of L. minutifolium bark five known metabolites were isolated using chromatographic techniques. The gastroprotective effects of the EtOAc fraction and edible infusion extract of the bark were assayed on the hydrochloric acid (HCl)/EtOH induced gastric ulcer model in mice to support the traditional use of the plant. The EtOAc extract and the edible infusion showed gastroprotective effect at dose of 100 mg/kg reducing lesions by 31% and 64%, respectively. The gastroprotective action mechanisms of the edible infusion at a single oral dose of 100 mg/kg were evaluated suggesting that prostaglandins, sulfhydryl groups, and nitric oxide are involved in the mode of gastroprotective action. The UHPLC analysis coupled to high-resolution mass spectrometry of the edible infusion showed the presence of twenty-three compounds. Our results can support the gastroprotective properties of the edible infusion extract, and at least can validate in part, the ethnopharmacological uses of the plant.

Dracocephalum moldavica L. Extracts Protect H9c2 Cardiomyocytes against H2O2-Induced Apoptosis and Oxidative Stress.

Materials and Methods The petroleum ether (petrol), dichloromethane (CH2Cl2), ethyl acetate (EtOAc), and n-butyl alcohol (n-BuOH) fractions were isolated from alcohol extracts of D. moldavica L. Total phenolic and flavonoid contents and in vitro antioxidant activities of different fractions were evaluated. H9c2 cells were then treated with D. moldavica L. extracts before challenging with H2O2. Cell viability was determined by colorimetric assay, and ELISA was used to measure the levels of lactate dehydrogenase (LDH), malondialdehyde (MDA), and superoxide dismutase (SOD). Apoptosis levels and mitochondrial membrane potential were measured by flow cytometry. The expressions of cell apoptosis regulatory proteins caspase-3, Bax, and Bcl-2 were determined by western blotting. Results Our results demonstrated that the EtOAc fraction from D. moldavica L. ethanol extract, which is rich in phenolic and flavonoid active constituents, had the strongest free radical scavenging activity. Additionally, this fraction increased H2O2-induced reduction in cell viability, SOD activity, and mitochondrial membrane potential. It also reduced H2O2-induced elevation in ROS production, contents of LDH and MDA, and H9c2 apoptosis. We further found that the EtOAc fraction increased Bcl-2 expression, while it decreased caspase-3 and Bax expressions induced by H2O2 in H9c2 cells. Conclusions Our data revealed that the EtOAc fraction from D. moldavica L. ethanol extract ameliorates H2O2-induced cardiotoxicity via antiapoptotic and antioxidant mechanisms.

Cultivated Fruit Body of Phellinus baumii: A Potentially Sustainable Antidiabetic Resource.

Previous studies have been reported that the fruit body of wild Phellinus baumii alleviates diabetes, and antioxidants are beneficial to diabetes by protecting the β-cell from damage due to oxidative stress. Large-scale cultivation of P. baumii fruit body has been successful in the past decade. This paper aimed to investigate whether the fruit body of the cultivated P. baumii has the same analogical effects as the wild. The cultivated P. baumii fruit body was extracted by 80% of ethanol extracts, and different fractions were obtained with the successive use of petroleum ether, ethyl acetate (EtOAc), n-butanol (n-BuOH), and water, which yielded 15.98 ± 1.56, 1.74 ± 0.34, 3.31 ± 0.41, 4.12 ± 0.37, and 1.38 ± 0.26% extract recovery, respectively. Results show that the EtOAc fraction exhibits the highest inhibitory effect on α-glucosidase activity (IC50 = 49.05 ± 3.14 μg mL–1), which is an order of magnitude higher than the positive control (acarbose, IC50 = 645.73 ± 7.86 μg mL–1). It was mainly composed of phenolic compounds with a purity of 79.45% and characterized by liquid chromatography-mass spectrometry as osmudacetone, hispidin, davallialactone, 2,5-bis(4,7-dihydroxy-8-methyl-2-oxo-2H-chromen-3-yl)cyclohexa-2,5-diene-1,4-dione, hypholomin B, and inoscavin A. Furthermore, the EtOAc fraction increased the glucose consumption of insulin-resistant HepG2 cells at a concentration range of 25–100 μg mL–1. The EtOAc fraction also demonstrated antioxidant activities by scavenging 1,1-diphenyl-2-picrylhydrazyl, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)diammonium salt, and hydroxyl radicals. In conclusion, the EtOAc fraction of the cultivated P. baumii fruit body exerted effective antidiabetic effects, possibly due to the high content of selective phenolic compounds. Hence, the cultivated fruit body of P. baumii can be a sustainable resource for treating diabetes, and our work also shed some light on its future utilization.

Anti-Adipogenic and Anti-Inflammatory Activities of (-)-epi-Osmundalactone and Angiopteroside from Angiopteris helferiana C.Presl.

Angiopteris helferiana C.Presl is a gigantic fleshy-type fern, belonging to Marattiaceae family. In previous study, we reported the potent anti-adipogenic and anti-inflammatory activities of ethylacetate (EtOAc) and n-butanol (BuOH) fractions of methanol extract of rhizomes of A. helferiana. In continuation, in this study, we report the isolation, characterization, and bioactivity analysis of principle bioactive compounds in these fractions. (−)-epi-Osmundalactone (1) and angiopteroside (2) were isolated from EtOAc and BuOH fractions, respectively. The structures of these compounds were established on the basis of NMR spectroscopic data. The quantification study using UPLC revealed the contents of compounds 1 and 2 in the dried rhizome to be 1.54% and 3.2%, respectively. These compounds were evaluated for their anti-adipogenic and anti-inflammatory activities using 3T3-L1 and RAW 264.7 cells, respectively. Compound 1 (2.5 µg/mL) and 2 (20 µg/mL) inhibited the lipid production by 35% and 25%, respectively. Regarding the anti-inflammatory activity, compound 1 (5 µg/mL) inhibited the nitrite production by nearly 82%. In conclusion, the presence of potent anti-adipogenic and anti-inflammatory compounds in A. helferiana indicate its potential role in the use of herb-based treatment for obesity and other related diseases.

Compound isolation and biological activities of Piptadeniastrum africanum (hook.f.) Brennan roots

Ethnopharmacological relevanceThe dicotyledonous plant Piptadeniastrum africanum (hook.f.) Brennan (Fabaceae) is used in traditional medicine to treat various human complaints including bronchitis, coughing, urino-genital ailments, meningitis, abdominal pain, treatment of wounds, malaria and gastrointestinal ailments, and is used as a purgative and worm expeller.Aim of the study: The present study describes the phytochemical investigation and the determination of the antimicrobial, antiplasmodial and antitrypanosomal activities of crude extract, fractions and compounds extracted from Piptadeniastrum africanum roots. Materials and methodsIsolated compounds were obtained using several chromatographic techniques. The structures of all compounds were determined by comprehensive spectroscopic analyses (1D and 2D NMR) and by comparing their NMR data with those found in literature. In vitro antimicrobial activity of samples was evaluated using the microdilution method on bacterial (Escherichia coli, Proteus mirabilis, Staphylococcus aureus) and fungal (Candida krusei) strains, while in vitro cell-growth inhibition activities were assessed against two parasites (Trypanosoma brucei brucei and Plasmodium falciparum strain 3D7). The cytotoxicity properties of samples were assayed against HeLa human cervical carcinoma. ResultsFive compounds were isolated and identified as: tricosanol 1, 5α-stigmasta-7,22-dien-3-β-ol 2, betulinic acid 3, oleanolic acid 4 and piptadenamide 5. This is the first report of the isolation of these five compounds from the roots of P. africanum. The (Hex:EtOAc 50:50) fraction exhibited moderate antibacterial activity against P. mirabilis (MIC 250 μg/mL), while the other fractions and isolated compounds had weak antimicrobial activities. Only the EtOAc fraction presented a moderate antimalarial activity with an IC50 of 16.5 μg/mL. The MeOH crude extract and three fractions (Hexane, Hexane-EtOAc 25% and EtOAc-MeOH 25%) exhibited significant trypanocidal activity with IC50 values of 3.0, 37.5, 3.8 and 9.5 μg/mL, respectively. ConclusionThese results demonstrated a scientific rational of the traditional uses of P. africanum and indicate that this plant should be further investigated to identify some of the chemical components that exhibited the activities reported in this study and therefore may constitute new lead candidates in parasiticidal drug discovery.

Investigation of Antioxidant and Elastase Inhibitory Activities of Geum urbanum Aerial Parts, Chemical Characterization of Extracts Guided by Chemical and Biological Assays

This study presents the bioguided chemical investigation of the 80% aqueous methanol extract of Geum urbanum aerial parts. Liquid–liquid partitioning of this extract in solvents of increasing polarity combined with biological screening showed that the ethyl acetate (EtOAc) soluble fraction was the most active part of the extract. This fraction was chemically profiled by a 13C nuclear magnetic resonance (NMR)-based dereplication method, resulting in the identification of 14 compounds. The dereplication process was followed by the purification of unknown and minor compounds of the EtOAc fraction. A new glycosylated phenol, namely, 3-(3,4-dihydroxyphenyl)propyl-α-l-rhamnopyranoside, together with 6 known compounds were isolated. Their structures were elucidated by spectroscopic methods including NMR and high-resolution electrospray ionization mass spectrometry. The antioxidant activity of fractions and isolated compounds were evaluated by 2,2,1-diphenyl-1-picrylhydrazyl and hydroxyl radical scavenging, and by cupric ion reducing antioxidant capacity assays. In parallel, their enzyme inhibitory property against human neutrophil elastase was assessed. Four subfractions, essentially containing polyphenols and triterpenes, exhibited a significant elastase inhibitory activity and an ellagitannin showed a very high radical scavenging activity.

Insecticidal activity of triterpenoids and volatile oil from the stems of Tetraena mongolica

Tetraena mongolica Maxim is a species of Zygophyllaceae endemic to China. Because few insect pests affect its growth and flowering, we speculated that this plant produces defensive chemicals that are insect repellents or antifeedants. The effects of different fractions from crude stem and leaf extracts on Pieris rapae were examined. The results confirmed that the ethyl acetate (EtOAc) fraction from the stems had insecticidal potential. Five compounds were isolated from the EtOAc fraction: a volatile oil [bis(2-ethylhexyl) benzene-1,2-dicarboxylate (1)], three triterpenoids 2E-3β-(3,4-dihydroxycinnamoyl)-erythrodiol (2), 2Z-3β-(3,4-dihydroxycinnamoyl)-erythrodiol (3), and 2E-3β-(3,4-dihydroxyphenyl)-2-propenoate (4)], and one steroid [β-sitosterol (5)]. Compounds 1–5 exhibited different degrees of insecticidal activity, including antifeedant and growth-inhibition effects. Compounds 1–5 inhibited the activity of carboxylesterase (CarE) and acetylcholinesterase (AChE) to different degrees. Compound 1 had the strongest antifeedant and growth-inhibition effects, and significantly inhibited the activity of CarE and AChE. Our results indicate that compounds 1–4 are the major bioactive insecticidal constituents of Tetraena mongolica. This work should facilitate the development and application of plant-derived botanical pesticides.

Anti-Inflammatory Effect of Flavonoids from Brugmansia arborea L. Flowers.

Brugmansia arborea L. (Solanaceae), commonly known as "angel's trumpet," is widely grown in North America, Africa, Australia, and Asia. It has been mainly used for ornamental purposes as well as analgesic, anti-rheumatic, vulnerary, decongestant, and anti-spasmodic materials. B. arborea is also reported to show anti-cholinergic activity, for which many alkaloids were reported to be principally responsible. However, to the best of our knowledge, a phytochemical study of B. arborea flowers has not yet been performed. Four flavonol glycosides (1-4) and one dihydroflavanol (5) were for the first time isolated from B. arborea flowers in this study. The flavonoids showed significant antioxidant capacities, suppressed nitric oxide production in lipopolysaccharide (LPS)-treated RAW 264.7 cells, and reduced inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX-2) protein production increased by LPS treatment. The contents of compounds 1-4 in n-BuOH fraction were determined to be 3.8 ± 0.9%, 2.2 ± 0.5%, 20.3 ± 1.1%, and 2.3 ± 0.4%, respectively, and that of compound 5 in EtOAc fraction was determined to be 12.7 ± 0.7%, by HPLC experiment. These results suggest that flavonol glycosides (1-4) and dihydroflavanol (5) can serve as index components of B. arborea flowers in standardizing anti-inflammatory materials.

Antioxidant and Antimicrobial Studies of Some Hemi-parasitic West African Plants

The study investigated the antioxidant and antimicrobial activities of some West African hemi-parasitic plants, which comprised Tapinanthus bangwensis (TB), Tapinanthus globiferus (TG) and Globimetula braunii (GB), used in Nigerian ethnomedicine for the management of skin and other microbial infections. This was with a view to determining the most active plant extract and fraction. The leaf and stem of each plant was separately air-dried, powdered and macerated in ethanol-H20 (8:2). The extracts were subjected to in vitro antioxidant tests such as 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP), Fe2+ chelating ability (FIC) and total antioxidant capacity (TAC) assays, with L-ascorbic acid, quercetin and ethylenediaminetetraacetic acid (EDTA) as positive controls. The antimicrobial test was carried out using micro-broth dilution method against reference strains of methicillin-resistant Staphylococcus aureus (MRSA), Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa and Candida albicans, where ciprofloxacin and ketoconazole were the positive controls. An activity-guided fractionation approach was adopted in the study. The most active GB leaf extract was fractionated to obtain n-hexane, dichloromethane, ethylacetate, n-butanol and aqueous fractions, and were subsequently tested. The results showed that the extract of GB leaf demonstrated the highest bioactivities with inhibitory concentration (IC50) of DPPH at 31.21±1.11 µg/mL, FRAP value of 109.30±0.76 mg AAE/g, TAC value of 178.15±3.54 mg AAE/g and MIC of 5.0 – 10.0 mg/mL. The Ethylacetate fraction of GB leaf demonstrated the highest bioactivities, which were four-times, thrice and twice better than its GB leaf extract in the DPPH, TAC and antimicrobial studies respectively. The EtOAc fraction (IC50 = 8.58±1.39 µg/mL) was comparable (P > 0.05) in antioxidant activity with quercetin (IC50 = 7.72±0.88 µg/mL). The EtOAc fraction also exhibited broad-spectrum antimicrobial activity with inhibitory concentration of 1.25 mg/mL against MRSA and C. albicans. In conclusion, the extract of G. braunii leaf demonstrated considerable bioactivities and the moderately polar EtOAc fraction was the most active fraction; hence, validates the folkloric use G. braunii leaf as a remedy for microbial infections.

Phytochemical Investigation, Antioxidant and Antimicrobial Activities of Ravenala Madagascariensis (Sonn.) Family Strelitziaceae Growing in Egypt

Introduction : Ravenala madagascariensis (Sonn.) family Strelitziaceae, commonly called traveler’s tree or palm; it is used traditionally in India as tea made from young leaves for treatment of diabetes. On the other hand its seed found to have antiseptic properties. Objective: Phytochemical investigation and isolation as well as testing the antioxidant and antimicrobial activities of the crude total and different successive fractions. Material and Method: The leaves total ethanol extract was subjected for phytochemical screening. The n- BuOH fraction was fractionated using chromatographic isolation. Total phenolics and flavonoids and antioxidant activity of the crude total ethanol extract as well as n-hexane, CH 2 Cl 2 , EtOAc and n- BuOH successive fractions were determined using ABTS assays. On the other hand the antimicrobial of the crude total ethanol was carried out against 13 microorganisms using agar well diffusion method. Results: Phytochemical screening showed the presence of flavonoids, tannins and sterols. Three phenolic compounds were isolated from the n- BuOH fraction and they were identified using different spectroscopic techniques. The different fractions showed high antioxidant activities especially EtOAc fraction that showed 88.96% for ABTS inhibition. Crude total ethanol extract showed selective moderate antimicrobial activity against the tested strains of Cryptococcus neoformans, Bacillus cereus and Serratia marcenscens. Conclusion: The ethyl acetate fraction showed significant antioxidant activity probably due to presence of the flavonoids contents

Systemic Administration of Calea pinnatifida Inhibits Inflammation Induced by Carrageenan in a Murine Model of Pulmonary Neutrophilia.

Objective The aim of this study was to investigate the anti-inflammatory effects of the crude extract (CE), derived fraction, and isolated compounds from Calea pinnatifida leaves in a mouse model of pulmonary neutrophilia. Methods The CE and derived fractions, hexane, ethyl acetate, and methanol, were obtained from C. pinnatifida leaves. The compounds 3,5- and 4,5-di-O-E-caffeoylquinic acids were isolated from the EtOAc fraction using chromatography and were identified using infrared spectroscopic data and nuclear magnetic resonance (1H and 13C NMR). Leukocytes count, protein concentration of the exudate, myeloperoxidase (MPO) and adenosine deaminase (ADA), and nitrate/nitrite (NOx), tumor necrosis factor-alpha (TNF-α), interleukin-1-beta (IL-1β), and interleukin-17A (IL-17A) levels were determined in the pleural fluid leakage after 4 h of pleurisy induction. We also analyzed the effects of isolated compounds on the phosphorylation of both p65 and p38 in the lung tissue. Results The CE, its fractions, and isolated compounds inhibited leukocyte activation, protein concentration of the exudate, and MPO, ADA, NOx, TNF-α, IL-1β, and IL-17A levels. 3,5- and 4,5-di-O-E-caffeoylquinic acids also inhibited phosphorylation of both p65 and p38 (P < 0.05). Conclusion This study demonstrated that C. pinnatifida presents important anti-inflammatory properties by inhibiting activated leukocytes and protein concentration of the exudate. These effects were related to the inhibition of proinflammatory mediators. The dicaffeoylquinic acids may be partially responsible for these anti-inflammatory properties through the inhibition of nuclear transcription factor kappa B and mitogen-activated protein kinase pathways.

Orostachys japonicus ethyl acetate fraction suppresses MRSA biofilm formation

Objective: To investigate the effect of Orostachys (O.) japonicus, a perennial herbaceous plant of the Family Crassulaceae, on biofilm formed by methicillin-resistant Staphylococcus aureus (MRSA). Methods: Powdered O. japonicus was extracted by 95% methanol, concentrated, and then, systematically fractionated with n-hexane, dichloromethane (DCM), ethyl acetate (EtOAc), n-butanol, and H2O according to polarity. Among them, the flavonoid-rich EtOAc fraction demonstrated the highest antibacterial activity and was used in this study. Using the biofilm inhibition assay, cell-surface attachment assay, confocal laser scanning microscopy, latex agglutination assay, and real time qRT-PCR, we examined whether the EtOAc fraction inhibited the formation of MRSA biofilm. Results: The EtOAc fraction exhibited distinct activity against biofilm formation and cell-surface attachment of MRSA up to 1 mg/mL through down-regulating the expression of mecA gene and the production and agglutination of penicillin-binding protein 2a as solidly observed in biofilm inhibition assay, cell-suface attachment assay, confocal laser scanning microscopy, latex agglutination assay, and real time qRT-PCR analysis. Conclusions: These results suggest that O. japonicus could be utilized as a potential resource for the development of new anti-biofilm formation of MRSA and antibacterial agents in the future.

Anwulignan from the fruits of Schisandra chinensis and its cytotoxicity against human cancer cell lines

Background: The fruits of Schisandra chinensis have been used traditionally as the medicinal materials in East Asia. Their extracts have been reported to be cytotoxic to several cancer cells and phytochemical research has identified numerous cytotoxic lignans. Objectives: We reviewed for the effects of cytotoxic lignans against various human cancer cells. Materials and Methods: Solvent extraction of fruits in aqueous methanol and fractionation of the extract into H2O, n-BuOH, and EtOAc fractions (Fr) were carried out. Column chromatography (CC) of the non-polar EtOAc Fr, which indicated cytotoxicity to cancer cells, was performed to isolate the cytotoxic lignin. They were identified on the basis of the intensive spectroscopic interpretation of IR, 1D-, 2D-NMR, and mass spectrometry data. The cytotoxicity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and cell staining using Hoechst 32258 and propidium iodide. Results: Anwulignan was isolated and identified as a cytotoxic principal component. The IC50 value was calculated to be 22.01 ± 1.87, 156.04 ± 6.71, and 32.68 ± 2.21 μM for human stomach adenocarcinoma, human colon cancer (HT29), and human cervical cancer cells, respectively. Conclusion: Solvent extraction, systematic fractionation, and repeated CC for S. chinensis fruits led to the isolation of a lignin identified to be anwulignan based on various spectroscopic analyses. Anwulignan demonstrated very high cytotoxicity to most human cancer cell lines, and cell death was induced by apoptosis.

Antisalmonellal Activities of Extracts, Fractions, Compounds and Semi-synthetic Flavonoid Derivatives from &amp;lt;i&amp;gt;Tristemma hirtum&amp;lt;/i&amp;gt; P. Beauv (Melastomataceae)

The development and spread of resistance to currently available antibiotics is a major drawback in the treatment of microbial infections. Salmonellosis for example remains among the most common cause of morbidity and mortality in developing countries. This study aimed to evaluate the antisalmonellal potential of extracts, fractions, isolated compounds and semi-synthetic flavonoids from <i>Tristemma hirtum</i> P. Beauv. Bioguided fractionation by column chromatography of the EtOAc and <i>n</i> -BuOH fractions led to the isolation of eleven compounds including two new esterified glucuronide flavonoids namely: luteolin-3′-<I>O</I>-<i>β</i>-<SUB>D</SUB>-glucuronopyranosylbutyl ester (1), a mixture of compound 1 and quercetin-3-<I>O</I>-<i>β</i>-<SUB>D</SUB>-glucuronopyranosylbutyl ester (2). Chemical transformation mainly based on the prenylation of 6-hydroxyapigenin-7-<I>O</I>-<i>β</i>-<SUB>D</SUB>-glucopyranoside (5) afforded four new semi-synthetic flavonoid derivatives namely: 6, 4'-<I>O</I>-diprenylapigenin-7-<I>O</I>-<i>β</i>-<SUB>D</SUB>-glucopyranoside (5a), 8-<I>C</I>-prenyl-6, 4'-<I>O</I>-diprenylapigenin-7-<I>O</I>-<i>β</i>-<SUB>D</SUB>-glucopyranoside (5b), 8-<I>C</I>-prenyl-4'-<I>O</I>-prenylapigenin-7-<I>O</I>-<i>β</i>-<SUB>D</SUB>-glucopyranoside (5c), 4'-<I>O</I>-prenylapigenin-7-<I>O</I>-<i>β</i>-<SUB>D</SUB>-glucopyranoside (5d). The chemical structures of these compounds were assigned using NMR techniques, mass spectrometry and by comparison of their data with reported ones. The antisalmonellal activity was assessed by determining the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) using serial microdilution methods. The results showed that the MeOH extract and EtOAc fraction were active against all the bacteria tested with MICs ranging from 24 to 1536 µg/mL. Seven isolated compounds and three semi-synthetic compounds tested showed MIC values ranging from 16 to 256 µg/mL. Compounds 1, 3, 5a, 5c and 11 displayed the most potent antisalmonellal properties but were generally less potent than those of reference drugs. The activity of extracts and isolated compounds could be used as the starting point for the development of alternative phytodrugs against salmonellosis.

A New Depsidone from Teloschistes flavicans and the Antileukemic Activity.

Lichens produce a variety of secondary metabolites that could be potential sources of pharmaceutically useful chemicals. However, only a limited number of lichen metabolites have been investigated for their biological significance. The objective of this study was to identify the potential compounds responsible for the antileukemic activity of lichen Teloschistes flavicans. Among three fractions (n-hexane, EtOAc, and MeOH-H2O), the ethyl acetate (EtOAc) fraction of T. flavicans methanolic extract showed the strongest inhibition in the HL-60 cell line. Additionally, the EtOAc fraction was further purified to obtain a new depsidone, 2,7-dichloro-3,8-dimethoxy-1,6,9-trimethyl-11H-dibenzo[b,e][1,4]dioxepin-11-one, named as flavicansone, along with rhizonic acid, parietin, and vicanicin. Flavicansone demonstrated the most significant inhibitory action against cell proliferation among the four isolated compounds.

Amelioration effects of Cirsium japonicum var. maackii extract/fractions on amyloid beta25-35-induced neurotoxicity in SH-SY5Y cells and identification of the main bioactive compound.

Amyloid beta (Aβ) is a neurotoxic peptide, and the accumulation of Aβ in the brain is the major characteristic of Alzheimer's disease (AD). Recently, the beneficial effects of Cirsium japonicum var. maackii (CJM) on brain health has attracted much attention. In the present study, we investigated the ability and protective mechanisms of CJM to attenuate neuronal toxicity caused by Aβ using SH-SY5Y cells. Aβ25-35 treatment decreased cell viability, whereas CJM extract/fractions increased cell viability in Aβ25-35-treated cells. We found that CJM treatment prevented the accumulation of reactive oxygen species observed in Aβ25-35-treated control cells. Furthermore, Aβ25-35-mediated production of inflammatory cytokines such as interleukin-1β was significantly suppressed by CJM. In addition, apoptotic factors were modulated in CJM-treated cells by downregulating B-cell lymphoma-2-associated X protein and upregulating B-cell lymphoma-2 protein expression. The assays showed that the ethyl acetate (EtOAc) fraction of CJM has greater neuroprotective bioactivities compared with the other extract/fractions. The main neuroprotective active compound from the EtOAc fraction of CJM was identified as pectolinarin using ultraperformance liquid chromatography-quadrupole time-of-flight-mass spectrometry. Collectively, this study not only describes the neuroprotective effect of CJM against Aβ25-35via the regulation of oxidative, inflammatory, and apoptotic signaling pathways, but also provides useful information for future studies on the mechanism of novel medicinal sources based on pectolinarin isolated from CJM.

A New Isoflavone and Other Constituents from Roots of Clitoria guianensis

A new isoflavone named pratensein-7-O-β-D-rutinoside [(-)-7-O-α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranosyl-5,3’-dihydroxy-4’-methoxyisoflavone] and the known compounds biochanin A-7-O-β-D-rutinoside, 6-deoxyclitoriacetal 11-O-β-D-glucopyranoside, 6-deoxyclitoriacetal, (2S)-naringenin-6-C-β-D-glucopyranoside, (2R)-naringenin-6-C-β-D-glucopyranoside, tachioside, and koaburaside were isolated from the roots of Clitoria guianensis (Aubl.) Benth var. guianensis (Fabaceae), a subshrub found in the Brazilian Cerrado biome. The structures of the compounds were identified by physical and spectroscopic data measurements (specific rotation ([α]D), circular dichroism (CD), ultraviolet (UV), infrared (IR), 1D and 2D nuclear magnetic resonance (NMR), and mass spectrometry (MS)). The EtOAc fraction of the roots exhibited high toxicity against Artemia salina with median lethal dose (LD50) value of 8.53 mg L-1.

Quercitrin, the Main Compound in Wikstroemia indica, Mitigates Skin Lesions in a Mouse Model of 2,4‐Dinitrochlorobenzene‐Induced Contact Hypersensitivity

Hapten-induced contact hypersensitivity (CHS) is widely utilized to induce immune activation in animal models of allergic contact dermatitis. Our previous findings suggested that the 95% EtOH extract of Wikstroemia indica (L.) C. A. Mey. has antiallergic and anti-inflammatory effects in DNCB-treated CHS SKH-1 hairless mice. The aim of this study was to evaluate the protective effects of compounds isolated from the EtOAc fraction of W. indica in RBL-2H3 cells and 2,4-dinitrochlorobenzene- (DNCB-) induced CHS mice. Of eight compounds in W. indica, that is, umbelliferone, daphnoretin, wikstrocoumarin, (+)-syringaresinol, tricin, (+)-lariciresinol, erythro-guaiacylglycerol-β-coniferyl ether, and quercitrin, quercitrin exhibited the most antiallergic activity against antigen-induced β-hexosaminidase release and IL-4 mRNA expression, which are markers of degranulation in RBL-2H3 cells. After a 7-sensitizing period, 14 days of DNCB treatment with or without topical pimecrolimus (1%) or quercitrin (0.5%) treatment, quercitrin was found to suppress DNCB-induced increases in serum IL-4 and IgE concentrations and transepidermal water loss. These results indicate that quercitrin has therapeutic potential for treatment of allergies and allergy-related contact dermatitis.

Chemical Investigation of Flavonoid, Phenolic acids Composition and Antioxidant activity of Tagetes erecta Flowers

This study was geared towards discovering the chemical profile and in vitro antioxidant activity of methanol (MeOH) extract and its ethyl acetate (EtOAc) fraction of Tagetes erecta orange flowers. A total of 29 known chemical compounds including phenolic acids and flavonoid compounds were identified and quantified using high performance liquid chromatography (HPLC) technique. The identification was carried out in EtOAc fraction for the first time. Pyrogallol and vanillic acid were highly abundant phenolic acids in flowers methanolic extract and its EtOAc fraction amounted 37.57 and 3.34mg/g dry extract, respectively. On the other hand, apig-6-rhamnose-8-glucose and hesperidin were found to be the major flavonoid compounds detected in MeOH extract amounted 18.34 and 0.66mg/g dry extract, respectively. Furthermore, results asserted that the tested samples were more efficient in the three applied antioxidant methods compared with standard antioxidant references (Ascorbic acid and BHT), especially EtOAc fraction which displayed the most activity against 1, 1-di phenyl-2-picrylhydrazyl (DPPH) radical with IC50 value 32.5± 0.12 µg/ml. These finding highlighted the use of Tagetes erecta flowers as a promising source of alternative bioactive compounds for both industrial and medicinal uses.