New pharmacological targets of three Asphodeline species using in vitro and ex vivo models of inflammation and oxidative stress.

This study explored the efficacy of the methanolic extract of three Asphodeline species (A. damascena subsp. rugosa, A. tenuior subsp. tenuiflora var. tenuiflora, and A. cilicica) to protect against hydrogen peroxide (H2O2)-induced lactate dehydrogenase (LDH) activity in HCT116 cells, and also any protective effects against lipopolysaccharides (LPS)-induced nitrite levels, prostaglandin E2 (PGE2) and 8-iso-prostaglandin F2α (8-iso-PGF2α) levels, 5HIAA/5-HT ratio, tumor necrosis factor (TNF)-α and interleukin (IL)-6 gene expression in rat colon specimens. Interestingly, A. tenuior extract was most effective in improving the tested biomarkers, by reducing LDH activity and nitrite level. On the other hand, A. damascena was the only species able to blunt LPS-induced TNF-α gene expression in rat colon specimens. The present findings highlighted the protective effects of Asphodeline extracts via in vitro and ex vivo models of inflammation and oxidative stress, adding new insights to the pharmacological actions of these medicinal plant species. Abbreviations: IBD: inflammatory bowel disease; LPS: lipopolysaccharide; LDH: lactate dehydrogenase; 5HIAA: 5-hydroxyindoleacetic acid; 5-HT: 5-hydroxytryptamine.

Effects of Kisspeptin-10 on Hypothalamic Neuropeptides and Neurotransmitters Involved in Appetite Control.

Besides its role as key regulator in gonadotropin releasing hormone secretion, reproductive function, and puberty onset, kisspeptin has been proposed to act as a bridge between energy homeostasis and reproduction. In the present study, to characterize the role of hypothalamic kisspeptin as metabolic regulator, we evaluated the effects of kisspeptin-10 on neuropeptide Y (NPY) and brain-derived neurotrophic factor (BDNF) gene expression and the extracellular dopamine (DA), norepinephrine (NE), serotonin (5-hydroxytriptamine, 5-HT), dihydroxyphenylacetic acid (DOPAC), and 5-hydroxyindoleacetic acid (5-HIIA) concentrations in rat hypothalamic (Hypo-E22) cells. Our study showed that kisspeptin-10 in the concentration range 1 nM⁻10 µM was well tolerated by the Hypo-E22 cell line. Moreover, kisspeptin-10 (100 nM⁻10 µM) concentration independently increased the gene expression of NPY while BDNF was inhibited only at the concentration of 10 µM. Finally, kisspeptin-10 decreased 5-HT and DA, leaving unaffected NE levels. The inhibitory effect on DA and 5-HT is consistent with the increased peptide-induced DOPAC/DA and 5-HIIA/5-HT ratios. In conclusion, our current findings suggesting the increased NPY together with decreased BDNF and 5-HT activity following kisspeptin-10 would be consistent with a possible orexigenic effect induced by the peptide.

Polyphenolic composition, enzyme inhibitory effects ex-vivo and in-vivo studies on two Brassicaceae of north-central Italy.

In this study, three different extracts (soxhlet, microwave and decoction) from two species of broccoli: Brassica oleracea L. convar. Italica botrytis (L.) Alef. var. cymosa Duch. (Broccolo Fiolaro) and Brassica oleracea acephala L. convar. acephala (DC.) Alef. var. sabellica L. (Cavolo Nero), which are commonly spread in north-central Italy, were tested for their enzyme inhibitory effects. Enzyme inhibitory effects were investigated against cholinesterases, tyrosinase, α-amylase and α-glucosidase. The soxhlet extracts had the highest inhibitory AChE effects with 1.08 mgGALAE/g (in Cavolo Nero) and 0.90 mgGALAE/g (in Broccolo Fiolaro). The significant tyrosinase inhibitory effect was observed in the soxhlet extract of Cavolo Nero with 11.93 mgKAE/g. In addition, we evaluated the antioxidant activity of Broccolo Fiolaro and Cavolo Nero on lipopolysaccharide (LPS)-stimulated bladder, kidney and liver specimens, ex vivo. We observed a significant reduction of both nitrite and malondialdehyde (MDA) following treatment that indicates a significant inhibitory effect on oxidative/nitrosative stress and lipoperoxidation, respectively. Additionally, the blunting effect induced by extracts on LPS-induced lactate dehydrogenase (LDH) activity further support a protective effect by both Broccolo Fiolaro and Cavolo Nero in bladder, kidney and liver. HPLC analysis revealed that catechin, epicatechin, vanillic and 3-hydroxy benzoic acids were the major components. The phenolic components may contribute to the observed enzyme inhibitory effects. in vivo tests also demonstrated that the extracts decreased the biochemical parameters in diabetic rats. Particularly, we observed the reduction of plasma glucose levels, urea and total cholesterol following oral administration, with the higher inhibitory effects exerted by Broccolo Fiolaro compared to Cavolo Nero. Overall, our results could provide new insights on the use of these Broccoli species not only as foods but also as functional and nutraceutical supplements.

Phytotherapic use of the Crocus sativus L. (Saffron) and its potential applications: A brief overview.

Crocus sativus L. (Saffron) has long been known for multiple target therapeutic uses. The plant metabolism is well investigated and the main metabolites related to saffron organoleptic qualities are crocin, crocetin, picrocrocin, and safranal. Particularly, the most abundant of them, such as crocin and safranal, are investigated for their multiple biological activities and known as potential drugs. We aimed to review the constituent features of the plant, along with its potential therapeutic effects in depression, neurodegenerative diseases, diabetes mellitus, atherosclerosis, cancer, and sexual dysfunction. A systematic literature search was conducted in PubMed, Medline, Scopus, and EMBASE, with particular attention to preclinical and clinical studies. Although saffron and its components showed potential clinical applications, further investigations are necessary to confirm the effective use of "Red Gold" and its real applications in clinical practice.

Graminex Pollen: Phenolic Pattern, Colorimetric Analysis and Protective Effects in Immortalized Prostate Cells (PC3) and Rat Prostate Challenged with LPS.

Prostatitis, a general term describing prostate inflammation, is a common disease that could be sustained by bacterial or non-bacterial infectious agents. The efficacy of herbal extracts with antioxidant and anti-inflammatory effects for blunting the burden of inflammation and oxidative stress, with possible improvements in clinical symptoms, is under investigation. Pollen extracts have been previously reported as promising agents in managing clinical symptoms related to prostatitis. The aim of the present work was to evaluate the protective effects of Graminex pollen (GraminexTM, Deshler, OH, USA), a commercially available product based on standardized pollen extracts, in rat prostate specimens, ex vivo. In this context, we studied the putative mechanism of action of pollen on multiple inflammatory pathways, including the reduction of prostaglandin E2 (PGE2), nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB), and malondialdehyde (MDA), whose activities were significantly increased by inflammatory stimuli. We characterized by means of chromatographic and colorimetric studies the composition of Graminex pollen to better correlate the activity of pollen on immortalized prostate cells (PC3), and in rat prostate specimens challenged with Escherichia coli lipopolysaccharide (LPS). We found that Graminex pollen was able to reduce radical oxygen species (ROS) production by PC3 cells and MDA, NFκB mRNA, and PGE2 levels, in rat prostate specimens. According to our experimental evidence, Graminex pollen appears to be a promising natural product for the management of the inflammatory components in the prostate.

Crocus sativus L. stigmas and byproducts: Qualitative fingerprint, antioxidant potentials and enzyme inhibitory activities.

Saffron (Crocus sativus L.) has been previously reported to be active as a protective agent in multiple experimental models of oxidative stress, inflammation and cancer. These findings refer to the protective effects of stigmas, not byproducts such as tepals and anthers. In this context, the aims of the present work were to characterize the phytochemical profile of saffron stigmas (CST) and high quality byproducts (tepals + anthers - CTA) extracts. Additionally, we studied the antioxidant and chelating effects of CST and CTA extracts by preliminary in vitro assay. The antioxidant activity was further investigated through the evaluation of reactive oxygen species (ROS) levels and lactate dehydrogenase (LDH) activity on mouse myoblast (C2C12) and human colon cancer (HCT116) cell lines. Additionally, we evaluated CST and CTA extract treatment on cholinesterases, α-glucosidase and α-amylase activity, in vitro. Finally, we studied the effects of CST extract on malondialdehyde (MDA) level in rat colon specimens challenged with E. coli lipopolysaccharide (LPS). We observed that water CST extracts are rich in phenolic content, whereas for CTA the olive oil was the elective extraction solvent. As expected, water CST extracts were the most effective in reducing hydrogen peroxide-induced oxidative stress in both cell lines and in vitro assays. Furthermore, both CST and CTA water extracts reduced the LDH activity in HCT116 cells challenged with hydrogen peroxide and LPS-induced MDA levels in rat colon specimens. Concluding, the present findings showed protective effects exerted by CST and CTA extracts in in vitro and ex vivo models of inflammation and oxidative stress.

A Comparative Assessment of Biological Effects and Chemical Profile of Italian Asphodeline lutea Extracts.

The present study aims to highlight the therapeutic potential of Asphodeline lutea (AL), a wild edible plant of the Mediterranean diet. Roots, aerial parts, and flowers of AL at two different phenological stages were collected from three locations in Italy. The inhibitory activities of extracts on strategic enzymes linked to human diseases were assessed. The antioxidant properties were evaluated in vitro, using six standard bioassays. The phenolic and anthraquinone profiles were also established using HPLC-PDA. Zinc, cadmium, lead, and copper contents were also determined. All the samples inhibited acetylcholinesterase (from 1.51 to 2.20 mg GALAEs/g extract), tyrosinase (from 7.50 to 25.3 mg KAEs/g extract), and α-amylase (from 0.37 to 0.51 mmol ACAEs/g extract). Aloe-emodin and physcion were present in all parts, while rhein was not detected. The phenolic profile and the heavy metals composition of specimens gathered from three different regions of Italy were different. It can be argued that samples collected near the street can contain higher concentrations of heavy metals. The experimental data confirm that the A. lutea species could be considered as a potential source of bioactive metabolites, and its consumption could play a positive and safe role in human health maintenance.

Protective Effects Induced by Microwave-Assisted Aqueous Harpagophytum Extract on Rat Cortex Synaptosomes Challenged with Amyloid ß-Peptide.

Harpagophytum procumbens is a plant species that displays anti-inflammatory properties in multiple tissues. The iridoid glycosides arpagoside, harpagide, and procumbide appear to be the most therapeutically important constituents. In addition, harpagoside treatment exerted neuroprotective effects both in vitro and in vivo. Considering these findings, the aim of the present work is to explore the possible protective role of the previously described microwave-assisted aqueous extract of H. procumbens on rat hypothalamic (Hypo-E22) cells, and in rat cortex challenged with amyloid ß-peptide (1-40). In this context, we assayed the protective effects induced by H. procumbens by measuring the levels of malondialdehyde, 3-hydroxykynurenine (3-HK), brain-derived neurotrophic factor, and tumor necrosis factor-α, 3-HK. Finally, we evaluated the effects of H. procumbens treatment on cortex levels of dopamine, norepinephrine, and serotonin. H. procumbens extract was well tolerated by Hypo-E22 cells and upregulated brain-derived neurotrophic factor gene expression but down-regulated tumor necrosis factor-α gene expression. In addition, the extract reduced amyloid ß-peptide stimulation of malondialdehyde and 3-HK and blunted the decrease of dopamine, norepinephrine, and serotonin, in the cortex. In this context, our work supports further studies for the evaluation and confirmation of Harpagophytum in the management of the clinical symptoms related to Alzheimer's disease. Copyright © 2017 John Wiley & Sons, Ltd.

Optimization of Aqueous Extraction and Biological Activity of Harpagophytum procumbens Root on Ex Vivo Rat Colon Inflammatory Model.

Harpagophytum procumbens has a long story of use for the treatment of inflammatory diseases. Considering both the antiinflammatory effects of H. procumbens in multiple tissues and the stability of harpagoside in artificial intestinal fluid, the aim of the present study was to explore the possible protective role of a microwave-assisted aqueous Harpagophytum extract (1-1000 µg/mL) on mouse myoblast C2C12 and human colorectal adenocarcinoma HCT116 cell lines, and isolated rat colon specimens challenged with lipopolysaccharide (LPS), a validated ex vivo model of acute ulcerative colitis. In this context, we evaluated the effects on C2C12 and HCT116 viability, and on LPS-induced production of serotonin (5-HT), tumor necrosis factor (TNF)-α, prostaglandin (PG)E2 and 8-iso-prostaglandin (8-iso-PG)F2α . Harpagophytum extract was well tolerated by C2C12 cells, while reduced HCT116 colon cancer cell viability. On the other hand, Harpagophytum extract reduced H2 O2 -induced (1 mM) reactive oxygen species (ROS) production, in both cell lines, and inhibited LPS-induced colon production of PGE2 , 8-iso-PGF2α , 5-HT and TNFα. Concluding, we demonstrated the efficacy of a microwave-assisted Harpagophytum aqueous extract in modulating the inflammatory, oxidative stress and immune response in an experimental model of inflammatory bowel diseases (IBD), thus suggesting a rational use of Harpagophytum in the management and prevention of ulcerative colitis in humans. Copyright © 2017 John Wiley & Sons, Ltd.

An Hydroalcoholic Chamomile Extract Modulates Inflammatory and Immune Response in HT29 Cells and Isolated Rat Colon.

Inflammatory bowel diseases (IBDs) are chronic disorders characterized by disruption and ulceration of the colonic mucosa or of any part of the digestive tract (Crohn's disease). Antioxidant/anti-inflammatory herbal extract supplementation could represent an innovative approach to contrast IBDs. Clinical trials demonstrated the efficacy of natural formulas, containing chamomile, in patients with gastrointestinal disorders. This is consistent, albeit in part, with the antioxidant and anti-inflammatory properties of chamomile. The aim of the present study was to explore the possible protective role of a chamomile extract, on human colorectal adenocarcinoma HT29 cell, and rat colon specimens treated with lipopolysaccharide (LPS) to induce an inflammatory stimulus, a well established model of acute ulcerative colitis. In this context, the activities of different biomarkers of inflammation and lipid peroxidation such as ROS, myeloperoxidase (MPO), serotonin (5-HT), prostaglandin (PG)E2 , 8-iso-prostaglandin (8-iso-PG)F2α , NF-kB, tumor necrosis factor (TNF)α and interleukin (IL)-6 were assessed. We found that chamomile extract was as effective as sulfasalazine (2 mg/ml) in reducing the production of MPO, 5-HT, IL-6, NF-kB, TNFα, PGE2 and 8-iso-PGF2α , after inflammatory stimulus. The observed modulatory effects support a rationale use of chamomile supplementation as a promising pharmacological tool for the prevention and management of ulcerative colitis in humans. Copyright © 2016 John Wiley & Sons, Ltd.

Antioxidant and antiproliferative activity of Stachys glutinosa L. ethanol extract.

Ethanol extracts of Stachys glutinosa L. (Lamiaceae) were investigated for antioxidative properties, as well as antiproliferative action on various cell lines. The antioxidant activities were investigated by ABTS (2,2'-azinobis-3-ethylbenzothiazoline-6-sulphonic acid) assay, DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging, ß-carotene/linoleic acid assay, scavenging of hydrogen peroxide (horseradish peroxidase test), superoxide anion scavenging, and hypochlorous acid scavenging (taurine test). The antioxidant activity was reported as IC50 and reveals antioxidant effects. Antiproliferative effects were measured in vitro on three cell lines: HepG2 (human hepatocarcinoma), MCF7 (breast human adenocarcinoma) and C2C12 (mouse myoblast) cell lines by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The ethanol extract induced variations in cell viability on all cell lines tested. At 200 µg/mL, the effects on cell viability were - 23%, - 27% and - 37%, respectively, for C2C12, MCF7 and HepG2.

Antioxidant activity of methanol extract of Helichrysum foetidum Moench.

Methanol extract of Helichrysum foetidum Moench (Asteraceae) was investigated for antioxidative properties. The antioxidant activities were investigated by 2,2'-azinobis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assay, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging, ß-carotene/linoleic acid assay, scavenging of hydrogen peroxide (HRPO test), superoxide anion scavenging (S.A.S. test) and hypochlorous acid scavenging (taurine test). The antioxidant activity was reported as IC50 and reveals Trolox-like antioxidative effects.

Chemical composition and inhibitory activity against Helicobacter pylori of the essential oil of Apium nodiflorum (Apiaceae).

The chemical composition of the essential oil obtained from Apium nodiflorum (L.) Lag. (Family Apiaceae), a plant used in the ethnomedical traditions of the Abruzzo region (Central Italy) as a culinary herb, as a diuretic, and to cure stomachache, was analyzed by gas chromatography/mass spectrometry, and 14 components were identified. Limonene (27.72 %), p-cymene (23.06%), myristicine (18.51%), and beta-pinene (6.62%) were the main components. The antimicrobial activity of the essential oil was assayed in vitro against Helicobacter pylori (strain DSMZ 4867), resulting in a minimum inhibitory concentration value of 12.5 microg/mL.

Phytochemical investigation on leaf extract of Cordia salicifolia Cham.

The dichloromethane extract of leaves of Cordia salicifolia Cham. (Family Boraginaceae) was fractionated by SiO(2) column chromatography and analyzed by gas chromatography, gas chromatography-mass spectrometry, and nuclear magnetic resonance spectroscopy. The apolar extract is characterized by a very high content of (+)-spathulenol (0.53%). The major component of the extract exhibited a very weak activity as an inhibitor of growth of Helicobacter pylori in vitro (minimum inhibitory concentration = 200 microg/mL).