Houttuynia cordata Thunb. Hexane fraction induces MDA-MB-231 cell apoptosis via caspases, ER stress, cell cycle arrest and attenuated Akt/ERK signaling.

Houttuynia cordata Thunb. (HCT) is a perennial plant used in traditional Thai medicine for many centuries. This study aimed to investigate the antiproliferative effect of the hexane fraction, which has not been explored before. HCT ethanol extract (crude extract) was sequentially fractionated to obtain a hexane (H) fraction. GC-MS was used to determine the phytochemicals. The H fraction consisted of lipids, mainly α-linolenic acid and some terpenoids. MTT assay was used to determine the cytotoxic effects of H fraction in MCF-7, MDA-MB-231, NIH3T3 and PBMCs. The mode of cell death and cell cycle analysis were determined by flow cytometry. The mechanisms of cell death were defined by mitochondrial transmembrane potential (MTP) reduction and activation of caspase-3, -8 and -9. The expression levels of the Bcl-2 family, cell cycle-related, endoplasmic reticulum (ER) stress-associated proteins; and Akt/ERK signaling molecules were investigated by immunoblotting. The H fraction was toxic to MDA-MB-231 more than MCF-7 cells but not to NIH3T3 and PBMCs. The growth of MDA-MB-231 cells was inhibited through apoptosis. MTP was disrupted whereas caspase-3, -8 and -9 were activated. The expression of pro-apoptotic Bax and Bak was upregulated, while Bid and anti-apoptotic Bcl-xL proteins were downregulated. Cyclin D1 and CDK4 levels were downregulated. The cell cycle was arrested at G1. Moreover, GRP78 and CHOP elevation indicated ER stress-mediated pathway. The expression ratio of pAkt/Akt and pERK/ERK were reduced. Taken together, the molecular mechanisms of MDA-MB-231 cell apoptosis were via intrinsic/extrinsic pathways, cell cycle arrest, ER stress and abrogation of Akt/ERK survival pathways. According to the most current research, the H fraction may be used as an adjuvant in the BC treatment; however, before the anticancer strategy can be applied to patients, it is important to determine each active compound's effects in cell lines and in vivo when compared with a combined mixture.

Hexane fraction of Globimetula braunii induces mitochondria-mediated apoptosis in Plasmodium berghei-infected mice.

Background: Apoptosis is a common pathology in malaria and most antimalarial drugs induce apoptosis during chemotherapy. Globimetula braunii is an African mistletoe used for the treatment of malaria but its effect on mitochondria-mediated apoptosis is not known. Methods: Malarial infection was induced by the intraperitoneal injection of NK 65 strain Plasmodium berghei-infected erythrocytes into mice which were treated with graded doses (100-400 mg/kg) of methanol extract (ME), and fractions of n-hexane, dichloromethane, ethylacetate and methanol (HF, DF, EF and MF) for 9 days after the confirmation of parasitemia. Artequine (10 mg/kg) was used as control drug. The fraction with the highest antiplasmodial activity was used (same dose) to treat mice infected with chloroquine-resistant (ANKA) strain for 5 consecutive days after the confirmation of parasitemia. P-alaxin (10 mg/kg) was used as control drug. On the last day of the treatment, liver mitochondria were isolated and mitochondrial Permeability Transition (mPT) pore opening, mitochondrial F0F1 ATPase (mATPase) activity, lipid peroxidation (mLPO) and liver deoxyribonucleic acid (DNA) fragmentation were assessed spectrophotometrically. Caspases 3 and 9 were determined by Enzyme-Linked Immunosorbent Assay (ELISA) technique. Cytochrome c, P53, Bcl-2-associated X protein (Bax), and B-cell lymphoma-2 (Bcl2) were determined via immunohistochemistry. Phytochemical constituents of the crude methanol extract of Globimetula braunii were determined via the Gas Chromatography-Mass Spectrometry (GC-MS) analysis. Results: There was large amplitude mPT induction by malaria parasites, extract and fractions of Globimetula braunii. At 400 mg/kg, HF significantly (p < 0.01) downregulated mATPase activity, and mLPO in both (susceptible and resistant) models, caused DNA fragmentation (P < 0.0001), induced caspases activation, P53, bax and cytochrome c release but downregulated Bcl2 in both models. The GC-MS analysis of methanol extract of Globimetula braunii showed that α-amyrin is the most abundant phytochemical. Conclusion: The n-hexane fraction of Globimetula braunii induced mitochondrial-mediated apoptosis through the opening of the mitochondrial pore, fragmentation of genomic DNA, increase in the levels of P53, bax, caspase 3 and 9 activation and cytochrome c release with concomitant decrease in the level of Bcl2. α-Amyrin is a triterpene with apoptotic effects.

In vitro effects and mechanisms of action of Bidens pilosa in Trypanosoma brucei.

Background and aim: African trypanosomiasis poses serious health and economic concerns to humans and livestock in several sub-Saharan African countries. The aim of the present study was to identify the antitrypanosomal compounds from B. pilosa (whole plant) through a bioactivity-guided isolation and investigate the in vitro effects and mechanisms of action against Trypanosoma brucei (T. brucei). Experimental procedure: Crude extracts and fractions were prepared from air-dried pulverized plant material of B. pilosa using the modified Kupchan method of solvent partitioning. The antitrypanosomal activities of the fractions were determined through cell viability analysis. Effects of fractions on cell death and cell cycle of T. brucei were determined using flow cytometry, while fluorescence microscopy was used to investigate alterations in cell morphology and distribution. Results and conclusion: The solvent partitioning dichloromethane (BPFD) and methanol (BPFM) fractions of B. pilosa exhibited significant activities against T. brucei with respective half-maximal inhibitory concentrations (IC50s) of 3.29 µg/ml and 5.86 µg/ml and resulted in the formation of clumpy subpopulation of T. brucei cells. Butyl (compound 1) and propyl (compound 2) esters of tryptophan were identified as the major antitrypanosomal compounds of B. pilosa. Compounds 1 and 2 exhibited significant antitrypanosomal effects with respective IC50 values of 0.66 and 1.46 µg/ml. At the IC50 values, both compounds significantly inhibited the cell cycle of T. brucei at the G0-G1 phase while causing an increase in G2-M phase. The results suggest that tryptophan esters may possess useful chemotherapeutic properties for the control of African trypanosomiasis.

Antidiarheal activity of catechol and ethyl 5, 8,11,14,17 - icosapentanoate-rich fraction of Annona senegalensis stem bark.

Background and aim: Secretory diarrhea is the most common type of diarrhea. This study aimed at exploring the possible mechanism of antisecretory action of Annona senegalensis stem bark and to identify the bioactive compounds. Experimental procedure: The ability of three crude extract; aqueous, dichloromethane and hexane stem bark extracts to inhibit castor oil-induced stooling in albino rats were assessed. Bioactivity guided fractionation of the most active extract was done using solvent-solvent partitioning (with hexane, dichloromethane, ethylacetate) and column chromatography. In vitro antioxidant activity of the most active sub-fraction was done using standard methods. The most active sub-fraction (25 mg/kg b. wt.) was administered to castor oil-induced diarrheal rats. Diarrheal rats small intestinal malondialdehyde concentration, antioxidant enzyme, cyclooxygenase II and Na+- K+ ATPase activities were determined using standard procedures. GC-MS analysis was done to identify the chemical compounds in the sub-fraction. Result and conclusion: Aqueous extract significantly decreased the number of wet stools. Sub-fraction 1 of ethylacetate fraction of aqueous stem bark extract (EFAS1) showed the highest stool inhibition. The H2O2 scavenging activity of EFAS1 was significantly greater than ascorbic acid. The sub-fraction significantly increased (p < 0.05) the activity of catalase and Na+- K+ ATPase activities but significantly decreased the concentration of malondialdehyde and cyclooxygenase II activity. GC-MS analysis revealed that EFAS1 is rich in catechol, n-hexadecanoic acid and ethyl-5,8,11,14,17-icosapentanoate. The sub-fraction exerts its antisecretory activity by its antioxidative, inhibition of prostaglandin synthesis and stimulation of Na+- K+ ATPase properties due to the presence of catechol, n-hexedecanoic acid and ethyl-5,8,11,14,17-icosapentanoate.

13, 14-Epoxyoleanan-3-ol-acetate: A Male Fertility-Enhancing Constituent from Hexane Fraction of Momordica charantia L. (Cucurbitaceae).

Objectives: Male infertility has been associated with oxidative stress-induced and/or microbial induced in some men. The use of medicinal plants to overcome oxidative stress-induced infertility cannot be over emphasized. Hence, the aim of this research was to isolate the antilipid peroxidation (an index of usage for treating oxidative stress-induced male infertility) bioactive principle from Momordica charantia using bioactivity-guided isolation. Materials and Methods: n-Hexane fraction from the crude ethanol extract obtained by Soxhlet extraction of aerial parts (without fruit) of bitter melon, M. charantia, was assessed for in vitro lipid peroxidation, followed by bioactivity-guided isolation of bioactive principles using in vitro lipid peroxidation as an index of aphrodisiac and male fertility enhancer. Results: Fractionation of the active n-hexane fraction using vacuum liquid chromatography (VLC) gave five pooled fractions on the basis of their thin layer chromatography (TLC) characteristics (n-hexane: EtOAc, 2:3, sulphuric acid spray). In vitro activity of the most active VLC fraction C was less than that of the positive control, vitamin E. Further fractionation of VLC-C by open column chromatography on silica gel led to the isolation of a compound which was purified by preparative-TLC. The purified compound, 10 mg/mL (Rf 0.54, TLC silica gel, n-hexane: ethyl acetate; 2:3) was equipotent with vitamin E (25 mg/mL) in reducing peroxidation of polyunsaturated fatty acids in vitro. Structural elucidation by NMR (1H, 13C) and mean mass spectroscopy confirmed the identity of the new bioactive compound as 13, 14-epoxyoleanan-3-ol-acetate. Conclusion: This study scientifically validates the traditional claim of M. charantia as an aphrodisiac or male fertility enhancer and suggests that 13, 14-epoxyoleanan-3-ol-acetate might be responsible for the observed activity.

Anxiolytic and antidepressant potential of extracts of Duchesnea Indica in animal models.

THIS study was designed to investigate the anxiolytic and antidepressant activity of Duchesnea Indica. The methanolic extract and n-hexane fraction of plant were tested in albino mice (20-30 g of 6-8 week). Safety profile of each extract was determined at different doses. Anxiolytic effect of plant was determined by Elevated Plus Maze and Light and Dark Model at different doses of methanolic and n-hexane extract. Antidepressant activities were assisted by Tail Suspension and Forced Swim Model at different doses. Result illustrated that D. Indica had a significant (P < 0.05) potential of reducing anxiety and depression. The methanolic extract at 100 mg and 200 mg significantly increased the time spend in light region of light and dark model and more time spent in open arm of Elevated Plus Maze model. n-hexane extract at dose of 5 mg and 10 mg/kg significantly increases the time spend in light region of Light and dark model and more time spend in open arm of Elevated Plus Maze model as compare to Control group. Methanolic extract of D. Indica significantly reduced the time of immobility in Tail Suspension and Force Swim Model at Dose of 100 mg and 200mg/kg. n-hexane extract also exhibit anti-depressant effect by reducing the time of immobility in Force swim and tail suspension Model at 100 mg and 200mg/kg dose as compare to control group. From the above observations, it could be assumed that the plant has marked anxiolytic and antidepressant potential. However further additional studies will be necessary to determine the underlying exact mechanism and clinical uses.

Extract, fractions, and ethyl-p-methoxycinnamate isolate from Kaempferia galanga Elicit anti-inflammatory activity by limiting leukotriene B4 (LTB4) production.

BACKGROUND AND AIM: Kaempferia galanga, also known as aromatic Ginger (kencur) in Indonesia, has been widely explored and shows potential as an anti-inflammatory agent. However, there has been limited research to show a possible mechanism by which aromatic ginger inhibits lipoxygenase (LOX). Therefore, this study aims to determine the anti-inflammatory activity of aromatic ginger by comparing extract, fractions, and ethyl-p-methoxycinnamate (EPMC) isolate, as well as possible LOX inhibition activity, by reducing the production of leukotriene B4 (LTB4). EXPERIMENTAL PROCEDURE: Two animal models were used, namely, the carrageenan-induced granuloma air pouch model and the pleurisy model. The test substance was administered 1 h before carrageenan induction, which was performed orally for each animal model. The number of leukocytes and the malondialdehyde (MDA) levels, leukotriene B4 (LTB4) levels, and histology were observed. GC-MS and LC-MS were used for analysis of the chemical compounds in the test samples. RESULTS AND CONCLUSION: The results of GC-MS analysis showed that aromatic ginger rhizome extract and fractions were dominated by ethyl-trans-p-methoxycinnamate, with the highest level found in the extract. K. galanga showed significant anti-inflammatory activity compared to the control (p < 0.01) in both the granuloma air pouch and pleurisy models. The results of examining the LTB4 concentration showed comparable activity between K. galanga extract, fractions and EMPC isolate, these results were not better than those of zileuton. Overall, this study shows that aromatic ginger extract, fractions and EPMC isolate have anti-inflammatory properties and have the potential to inhibit LOX, thereby reducing LTB4 levels.

The potentiation of beta-lactam and anti-bacterial activities of lipophilic constituents from Mesua ferrae leaves against methicillin-resistant Staphylococcus aureus.

OBJECTIVES: Mesua ferrae, from the family of Calophyllaceae, is traditionally used for the treatment of piles, fever and renal disorders. The present study was aimed to examine the antibacterial compounds from the leaves of M. ferrae and their ß-lactam antibiotic potentiate activities against Staphylococcus aureus and methicillin-resistant Staphylococcus aureus (MRSA). METHODS: Stigmasterol (1) and ß-caryophyllene oxide (2) were isolated from the n-hexane fraction of the leaves of M. ferrae using a bioassay-guided fractionation approach. RESULTS: The isolated compounds displayed anti-Staphylococcus and anti-MRSA activities. It is worth to note that both compounds demonstrated synergism with ß-lactam antibiotics against S. aureus and MRSA. Gas chromatography-mass spectrometry (GC-MS) analysis indicated the n-hexane fraction was dominated by triterpenes and sesquiterpenes, suggesting the total antibacterial activity exhibited by the fraction. CONCLUSION: Based on the findings, it could conclude that M. ferrae is a promising natural source for the discovery of new anti-MRSA lead compounds.

Bio-guided isolation of anti-leishmanial natural products from Diospyros gracilescens L. (Ebenaceae).

BACKGROUND: Plants represent an intricate and innovative source for the discovery of novel therapeutic remedies for the management of infectious diseases. The current study aimed at discovering new inhibitors of Leishmania spp., using anti-leishmanial activity-guided investigation approach of extracts from Diospyros gracilescens Gürke (1911) (Ebenaceae), targeting the extracellular (promastigotes) and intracellular (amastigotes) forms of Leishmania donovani. METHODS: The plant extracts were prepared by maceration using H20: EtOH (30:70, v/v) and further fractionated using a bio-guided approach. Different concentrations of D. gracilescens extracts, fractions and isolated compounds were tested in triplicate against L. donovani promastigotes and amastigotes in vitro. The antileishmanial potency and cytotoxicity on RAW 264.7 cells were determined using the resazurin colorimetric assay. The time kill kinetic profile of the most active sample was also investigated. The structures of all compounds were elucidated on the basis of extensive spectroscopic analyses, including 1D and 2D NMR, and HR-ESI-MS and by comparison of their data with those reported in the literature. RESULTS: The hydroethanolic crude extract of D. gracilescens trunk showed the most potent antileishmanial activity (IC50 = 5.84 µg/mL). Further fractionation of this extract led to four (4) fractions of which, the hexane fraction showed the most potent activity (IC50 = 0.79 µg/mL), and seven (07) compounds that exhibited moderate potency (IC50 = 13.69-241.71 µM) against L. donovani. Compound 1-deoxyinositol (7) inhibited the promastigote and amastigote forms of L. donovani with IC50 values of 241.71 µM and 120 µM respectively and also showed the highest selectivity against L. donovani promastigotes (SI > 5.04). To the best of our knowledge, the antileishmanial activity of this compound is being reported here for the first time. The promising hexane fraction showed significant inhibition of parasites growth at different concentrations, but with no evidence of cidal effect over an exposure period of 120 h. CONCLUSIONS: The results obtained indicated that the hydroethanolic extract from the D. gracilescens trunk and the derived hexane fraction have very potent inhibitory effect on cultivated promastigotes and amastigotes of L. donovani parasite. The isolated compounds showed a lesser extent of potency and selectivity. However, further structure-activity-relationship studies of 1-deoxyinositol could lead to more potent and selective hit derivatives of interest for detailed drug discovery program against visceral leishmaniasis.

Non-Polar Myxococcus fulvus KYC4048 Metabolites Exert Anti-Proliferative Effects via Inhibition of Wnt/ß-Catenin Signaling in MCF-7 Breast Cancer Cells.

The Wnt/ß-catenin signaling pathway is involved in breast cancer and Myxococcus fulvus KYC4048 is a myxobacterial strain that can produce a variety of bioactive secondary metabolites. Although a previous study revealed that KYC4048 metabolites exhibit anti-proliferative effects on breast cancer, the biochemical mechanism involved in their effects remains unclear. In the present study, KYC4048 metabolites were separated into polar and non-polar (ethyl acetate and n-hexane) fractions via liquid-liquid extraction. The effects of these polar and non-polar KYC4048 metabolites on the viability of breast cancer cells were then determined by MTT assay. Expression levels of Wnt/ß-catenin pathway proteins were determined by Western blot analysis. Cell cycle and apoptosis were measured via fluorescence-activated cell sorting (FACS). The results revealed that non-polar KYC4048 metabolites induced cell death of breast cancer cells and decreased expression levels of WNT2B, ß-catenin, and Wnt target genes (c-Myc and cyclin D1). Moreover, the n-hexane fraction of non-polar KYC4048 metabolites was found most effective in inducing apoptosis, necrosis, and cell cycle arrest, leading us to conclude that it can induce apoptosis of breast cancer cells through the Wnt/ß-catenin pathway. These findings provide evidence that the n-hexane fraction of non-polar KYC4048 metabolites can be developed as a potential therapeutic agent for breast cancer via inhibition of the Wnt/ß-catenin pathway.

Anti-toxoplasma, antioxidant and cytotoxic activities of Pleopeltis crassinervata (Fée) T. Moore hexane fraction.

The apicomplexan parasite Toxoplasma gondii (T. gondii) causes toxoplasmosis in humans. Pyrimethamine and sulfadiazine that are the drugs of choice to treat the disease, produce severe side effects as well as failure treatments because of drug resistance; thus, novel anti-Toxoplasma compounds are needed and natural compounds can be a good source to obtain them, as medicinal plants have been used to control other apicomplexan parasites. Pleopeltis crassinervata (P. crassinervata) is a fern used in some rural areas of Mexico to treat among other malaises, mouth ulcers, gastrointestinal problems and parasites. Therefore, the efficacy of extracts and fractions obtained from P. crassinervata fronds was evaluated on the viability of T. gondii RH strain tachyzoites by the Stytox green method. RH is the prototypical type 1 Toxoplasma strain, isolated for the first time from the brain of a patient boy named R. H. Its phytochemical profile, MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, Hep-2 cytotoxicity and antioxidant activity by ABTS (2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) and DPPH (2,2-diphenyl-1-picrylhydrazyl) methods, were also assessed. Hexane fraction exhibited the highest anti-Toxoplasma activity with an IC50 of 16.90 µg/mL. This fraction did not show antioxidant activity and contained at least 2 terpenoid type compounds with retention factor (Rf) of 0.75 and 0.86. The fraction was not toxic to the host cells in doses up to 50 µg/mL. P. crassinervata frond hexane fraction seems to be a good candidate to obtain possible anti-Toxoplasma compounds. This study is the first to report the biological, antioxidant and cytotoxic activity of P. crassinervata fern.

Hypnotic Effect of Portulaca oleracea on Pentobarbital-Induced Sleep in Mice.

OBJECTIVE: In Iranian Traditional Medicine, the herbs with cold and wet temperament can help to improve insomnia. Portulaca oleracea has a cold and wet temperament, so the present study was carried out to investigate the sleep-prolonging effect of Portulaca oleracea. METHODS: This work was an experimental study on mice which were randomly divided into these groups: saline (control); Diazepam: positive control); hydro-alcoholic extract of Portulaca oleracea (12.5, 25, 50, 75 and 100 mg/kg) by Soxhlet apparatus and maceration; in the effective (dose25 mg/kg), different fractions of extract were tested. Ethyl acetate fraction (EAF:); N hexane fraction (n-HF); water fraction (WF). All the test compounds were injected intraperitoneally (IP) 30 minutes before pentobarbital administration (30 mg/kg). Duration and latency of pentobarbital-induced sleep were recorded. Also, LD50 of Portulaca oleracea extract was determined and the possible neurotoxicity of the extract was tested on neural PC12 cells. Besides, 30 min after administration of hydroalcoholic extract (HAE) motor coordination (rota-rod test) was assessed. RESULTS: HAE increased the duration of pentobarbital-induced sleep at doses of 25, 50, 75 and 100 mg/kg. The hypnotic effect of HAE was comparable to that induced by diazepam. Similarly, WF, EAF, and NHF at 25 mg/kg could increase sleep duration. The sleep latency was decreased by HAE and NHF but not by WF and EAF. The LD50 value for HAE was found to be 4.8 g/Kg. HAE and its fractions did not show neurotoxic effect in cultured PC12-cell line, also HAE did not affect the animals performance on the rotarod test. CONCLUSION: The present data demonstrated that Portulaca oleracea potentiates sleeping behaviors. The main components responsible for the hypnotic effects of this plant is most likely a non-polar agents which is found in NHF. Isolation of the active constituents may yield a novel sedative drug.

The cytotoxic nature of Acanthopanax sessiliflorus stem bark extracts in human breast cancer cells.

Acanthopanax sessiliflorus, a small woody shrub has traditionally been referred to have anticancer activity, but it has not been scientifically explored so far. Therefore, to investigate the anticancer effects of A. sessiliflorus stem bark extracts (ASSBE), MDA-MB-231 and MCF-7 human breast cancer cells were treated with one of its bioactive fractions, n-hexane (ASSBE-nHF). Cytotoxicity (24 h) was determined by MTT assay and antiproliferative effect was assessed by counting cell numbers after 72 h treatment using hemocytometer. The role of ASSBE-nHF on apoptosis was analysed by annexin V-FITC/PI, Hoechst 33342 staining, DNA fragmentation pattern and immunoblotting of apoptosis markers. For the assay of enhanced production of ROS and mitochondrial membrane depolarization, specific stains such as DCFH-DA and JC-1 were used, respectively. To understand the mode of action of ASSBE-nHF on MCF-7 cells, cells were pre-treated with antioxidant, n-acetylcysteine. The hexane fraction of ASSBE showed maximum activity towards human breast cancer cells compared to other two fractions at a minimal concentration of 50 µg/ml. The annexin V-FITC/PI, Hoechst 33342 staining, DNA fragmentation and immunoblotting assays showed that ASSBE-nHF induces non-apoptotic cell death in MCF-7 and MDA-MB-231 cells. ASSBE-nHF significantly increased the production of ROS and decreased the mitochondrial membrane potential (MMP) in MCF-7 cells. Similarly, it decreased the MMP in MDA-MB-231 cells, but had no effect on ROS production. Further, the cytotoxic effect of ASSBE-nHF in MCF-7 cells was not significantly reversed even in the presence of n-acetylcysteine, an antioxidant. These findings revealed that ASSBE-nHF induces non-apoptotic cell death via mitochondria associated with both ROS dependent and independent pathways in human breast cancer cells.

Cytotoxic agents for KB and SiHa cells from n-hexane fraction of Cissampelos pareira and its chemical composition.

Eleven constituents were characterised by gas chromatography-mass spectrometry analysis, and five molecules were isolated using column chromatography. The in vitro study of the extract and isolated molecules against KB and SiHa cell lines revealed oleanolic acid (1) and oleic acid (2) as potent cytotoxic molecules with potential anticancer activity. The IC50 values of n-hexane extract (CPHF), oleanolic acid (1) and oleic acid (2) were >300, 56.08 and 70.7 µg/mL (µM), respectively, against KB cell lines and >300, 47.24 and 80.2 µg/mL (µM), respectively, against SiHa cell lines.

Anti-inflammatory activity of methanol extract and n-hexane fraction mojabanchromanol b from Myagropsis myagroides.

AIMS: This study was carried out to verify the anti-inflammatory effect of methanol extract from Myagropsis myagroides (MMME) and its n-hexane fraction mojabanchromanol b. MAIN METHODS: The murine macrophages Raw264.7 cells were used. The pro-inflammatory cytokines (IL-6, IL-1ß, TNF-α) and the expression of iNOS, COX-2, and NF-κB p65 were examined by ELISA and immunoblotting. To investigate the inhibitory effect of MMME in an animal model of inflammation, an assay to determine croton oil-induced ear edema in mice was performed. KEY FINDINGS: NO levels decreased with increasing concentration of MMME, and were inhibited up to 50%. The secretion of IL-6, TNF-α, and IL-1ß was suppressed in a dose-dependent manner, especially at 50µg/mL, inhibition activities of cytokines were over 50%. MMME also suppressed the expression of COX-2, iNOS, and NF-κB p65, suggesting that MMME could affect the expression of inflammation related cytokines and proteins through the deregulation of NF-κB. Moreover, the formation of mouse ear edema was reduced at the highest dose tested compared to that in the control, and generated similar effects compared with prednisolone at 250mg/kg in mice ear edema evaluation test. In addition, the results in photomicrograph of mice ear tissue and mast cells also showed the same effect. After purification of fractions of MMME, it indicated that n-hexane fraction mojabanchromanol b was the most active fraction showing the inhibitory effect of IL-6 and TNF-α. SIGNIFICANCE: These results suggested that MMME and mojabanchromanol b may have great effects on inflammatory factors and be potential anti-inflammatory therapeutic materials.

Phytochemical study guided by the myorelaxant activity of the crude extract, fractions and constituent from stem bark of Hymenaea courbaril L.

ETHNOPHARMACOLOGICAL RELEVANCE: Hymenaea courbaril L. (Caesalpinoideae) is used in Brazilian folk medicine to treat anemia, kidney problems, sore throat and other dysfunctions of the respiratory system, such as bronchitis and asthma, although such properties are yet to be scientifically validated. AIM OF THE STUDY: In order to give a scientific basis to support the traditional use of Hymenaea courbaril, this study was designed to evaluate antioxidant, myorelaxant and anti-inflammatory properties of the ethanol extract from stem bark and its fractions. The myorelaxant effect of astilbin, a flavonoid isolated from the bioactive ethyl acetate fraction (EAF), has also been evaluated. MATERIAL AND METHODS: In the present study ethanol extract from stem bark (EEHC) and fractions were analyzed using bioassay-guided fractionation. The following activities were investigated: antioxidant by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, myorelaxant on rat tracheal smooth muscle, and anti-inflammatory using ovalbumin-induced leukocytosis and airway hyperresponsiveness in rats. RESULTS: The results of the present investigation show that the whole extract of Hymenaea courbaril and some of its fractions strongly scavenged DPPH radical. The extract showed myorelaxant activity on rat trachea, being EAF its highest efficient fraction. Bio-guided study allowed the isolation of astilbin, a well-known flavonoid. The activity induced by this compound indicates that it may be partly responsible for the myorelaxant effect of EAF. EAF reduced contractions that depended on divalent cation inflow through voltage-operated Ca(2+) channels (VOCCs) or receptor-operated Ca(2+) channels (ROCCs), but it was more potent to inhibit VOCC- than ROCC-dependent contraction induced by Ca(2+) addition in ACh-enriched Ca(2+)-free medium. Oral pretreatment of antigen-challenged animals with EAF prevented airway hyperresponsiveness on KCl-induced contraction and reduced the number of total white cells, particularly eosinophils and neutrophils in bronchoalveolar lavage. CONCLUSIONS: This study provided scientific basis that Hymenaea courbaril presents potential antioxidant, myorelaxant and anti-inflammatory actions, which support its use in folk medicine to treat inflammatory airway diseases.

Citotocixidade e atividade antiparasitária de Lygodium venustum SW / Cytotoxic and antiparasitic activity of Lygodium venustum SW

. Doenças parasitárias infecciosas como leishmaniose e doença de Chagas tem se difundido nas últimas décadas a locais onde antes não se observava sua ocorrência. São consideradas negligenciadas por assolarem países pobres e serem marginalizadas farmacologicamente. O tratamento não apresenta muitas opções de fármacos e estes demonstram relevante toxicidade contribuindo para o aparecimento de diversos efeitos colaterais. A pesquisa com produtos naturais tem se mostrado uma interessante alternativa para a procura por novos fármacos. Lygodium venustum é uma samambaia cosmopolita de hábito lianescente encontrada na encosta na Chapada do Araripe, considerada por algumas populações americanas como planta medicinal para o tratamento de dermatoses, infecções, micoses e tricomoníases. Neste estudo foi avaliada sua atividade anti-parasitária contra Leishmania brasiliensis e Trypanosoma cruzi, bem como sua citotoxicidade através de ensaios n vitro. Foram testadas a fração hexânica e o extrato etanólico obtido das folhas de Lygodium venustum em diferentes concentrações. Para os testes in vitro de T. cruzi, foi utilizado o clone CL-B5 e para Leishmania brasiliensis foram utilizadas formas promastigotas. O ensaio de citotoxicidade foi realizado com linhagens de fbroblastos. L. venustum não apresentou atividade antiparasitária clinicamente relevante na forma de extrato etanólico bruto nem como fração hexânica contra Leishmania. A fração hexânica apresentou uma atividade intermediária contra T. cruzi, porém a concentração de efeito moderado possui citotoxicidade máxima tornando-se inviável para aplicação clínica. Entretanto, a citoxicicidade apresentada poderá ser útil em pesquisas sobre atividade antineoplásica em células tumorais.

Infectious and parasitic diseases like leishmaniasis and Chagas disease have spreading recent decades to places not observed before. They are considered neglected by desolating poor countries and marginalized pharmacologically. There are not many options for the treatment and these drugs have shown signifcant toxicity contributing to the appearance of several side effects. Research on natural products has been shown to be an interesting alternative to the search for new drugs. Lygodium venustum is a cosmopolitan fern with latescence habit found on the Chapada do Araripe, considered by some American popula-tions as a medicinal plant for the treatment of skin diseases, infections, fungal infections and trichomoniasis. This study evaluated its antiparasitic activity against Trypanosoma cruzi and Leishmania brasiliensis, as well as its cytotoxicity through trials in vitro. We tested the ethanolic extract and hexane fraction obtained from the leaves of L. venustum at different concentrations. For in vitro tests of T. cruzi, we used the clone CL-B5 and for L. brasiliensis we used promastigotes. The cytotoxicity assay was performed with strains of fbroblasts. L.venustum showed no antiparasitic activity clinically relevant in the form of crude ethanolic extractor as the hexane fraction against Leishmania. The hexane fraction showed an intermediate activity against T.cruzi, but the concentration of moderate effect has maximum cytotoxicity becoming unfeasible for clinical application. However, the cytotoxicity presented may be useful in research on antineoplastic activity in tumor cells.

Antipyretic and anticonvulsant activity of n-hexane fraction of Viola betonicifolia.

OBJECTIVE: To investigate the antipyretic and anticonvulsant activities of n-hexane fraction of Viola betonicifolia (V. betonicifolia). METHODS: The antipyretic effect was scrutinized using brewer's yeast induced pyrexia and anticonvlsion effect was tested using pentylenetetrazol and strychnine induced convulsion in mice. RESULTS: N-hexane fraction of V. betonicifolia demonstrated highly significant antipyretic activity during various assessment times (1-5 h) when challenged in yeast induced pyrexia test. The effect was in a dose dependent manner with maximum attenuation (82.50%) observed at 300 mg/kg i.p. When tested in pentylenetetrazol induced convulsion test, the 1st stage (Ear and facial twitching) and 2nd stage (Convulsive wave through the body) was 100% protected during 24 h at all the test doses (300, 400 and 500 mg/kg i.p.), while the latency time of remaining stages was significantly increased. The maximum effect was observed by n-hexane fraction of V. betonicifolia at 400 and 500 mg/kg i.p., as the latency time for generalized clonic-tonic seizure (5th stage) was increased up to 25.34 min. However, n-hexane fraction of V. betonicifolia had no protection in strychnine induced convulsion test. CONCLUSIONS: In conclusion, phytopharmacological studies provide scientific foundation to the folk uses of the plant in the treatment of pyrexia and neurological disorders.

Herbs in association with drugs: Enhancement of the aminoglycoside-antibiotic activity by Pityrogramma calomelanos (L.) Link.

AIMS: In this work, the ethyl acetate and hexane fractions of Pityrogramma calomelanos (L.) were evaluated to antibacterial and antifungal activity against strains of Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans, Candida krusei and Candida tropicalis. METHODS: The study was performed aiming to assess the antimicrobial effect with the method of dilution in HIA. The hexane and ethyl acetate fractions were named HFPC and EAFPC, respectively. RESULTS: Both fractions of specie P. calomelanos displayed good activity against S. aureus when associated with gentamicin. When associated with the antifungal, the fractions did not exhibit relevant activity against species of Candida. CONCLUSIONS: These results indicate that this specie can be used as a possible source of natural products of antibacterial interest, mainly when combined with aminoglycosides.