Luigi Menghini

Antiinflammatory effects in THP-1 cells treated with verbascoside

Verbascum thapsus commonly known as ‘mullein’ is part of a large family of Scrophulariaceae consisting of more than 360 species. From antiquity Verbascum thapsus has been used as a medicinal herb, it contains diverse polysaccharides, iroid glycosides, flavonoids, saponins, volatile oils and phenylentanoids. Inducible nitric oxide synthase (iNOS) represents one of the three isoforms that produce nitric oxide using L‐arginine as a substrate in response to an increase in superoxide anion activated by NF‐kB. It is implicated in different pathophysiological events and its expression increases greatly during an inflammatory process, due to oxidative stress and the activation of the enzymes of the antioxidant network such as SOD, CAT and GPx.

Comparison of three different extraction methods and HPLC determination of the anthraquinones aloe-emodine, emodine, rheine, chrysophanol and physcione in the bark of Rhamnus alpinus L. (Rhamnaceae)

INTRODUCTION Rhamnus alpinus L. (Rhamnaceae), a traditional plants in the flora of the Abruzzo region, is known to contain active anthraquinone secondary metabolites. However, the content of anthraquinones varies among R. alpinus samples depending on collection season and site. Thus, using simple, reliable and accurate analytical methods for the determination of anthraquinones in R. alpinus extracts allows comparative study of different methods of extraction. OBJECTIVE After a partial validation of an HPLC method for the simultaneous determination of five anthraquinones, aloe-emodine, rheine, emodine, chrysophanol and physcione, in the bark of R. alpinus, we compared three different methods of extraction. METHODOLOGY Anthraquinones were extracted from the bark of R. alpinus using different techniques (methanol maceration, ultrasonic and supercritical CO(2) extraction). Separation and quantification of anthraquinones were accomplished using a reversed-phase C(18) column with the mobile phase of H(2)O-methanol (40 : 60, v/v, 1% formic acid) at a wavelength of 254 nm. The qualitative analyses were also achieved at wavelength of 435 nm. RESULTS All calibration curves were linear over the concentration range tested (10-200 mM) with the determination coefficients >or=0.991. The detection limits (S/N = 3) were 5 mM for each analytes. All five anthraquinones were found in the samples tested at concentrations reported in experimental data. CONCLUSION The described HPLC method and optimised extraction procedure are simple, accurate and selective for separation and quantification of anthraquinones in the bark of R. alpinus and allow evaluation of the best extraction procedure between the tested assays.

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 (Crohns 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

Comparative study of biological activities and multicomponent pattern of two wild Turkish species: Asphodeline anatolica and Potentilla speciosa.

Abstract The multicomponent pattern and biological characterization of plant material are essential for pharmaceutical field, in the food supplements quality control procedures and to all plant-based products. These nutrients often show valuable effects related to their consumption due to the occurrence of secondary metabolites that show useful properties on health. In this framework, researches performed on this topic play a central role for human health and drug development process. The aim of this study was to compare phenolics and free anthraquinones multicomponent pattern of two wild Turkish species: Asphodeline anatolica and Potentilla speciosa using validated high-performance liquid chromatography–photogiode array (HPLC–PDA) assays, coupled to biological evaluation. Even if some variances related to biological and enzymatic inhibition activities can be ascribed to other phytochemicals, the reported data support traditional use of Asphodeline anatolica and Potentilla speciosa roots as valuable natural font for the development of novel natural-derived drug formulations and/or food supplements with health and nutritional benefits.

Antioxidant Effects of Garlic in Young and Aged Rat Brain In Vitro

Oxidative stress plays a key role in the pathogenesis of age-related neurodegeneration, and the nonenzymatic production of 8-iso-prostaglandin F(2alpha) (8-iso-PGF(2alpha)) may represent a reliable index of cellular oxidative damage. Garlic (Allium sativum) has been associated with peripheral antioxidant activities and therefore might prevent or reverse 8-iso-PGF(2alpha) production, but scant data are available on its possible neuroprotective effects. Therefore, we have studied the possible antioxidant effects of a garlic extract in rat brain synaptosomes obtained from young (3-month-old) and aged (14-month-old) male Wistar rats that were perfused, in vitro, with graded concentrations of a garlic extract (10-500 microg/mL). Release in the effluent was evaluated, both in the basal state and after hydrogen peroxide-induced oxidative stress. In young rats, we observed a concentration-dependent inhibitory effect of the garlic extract on brain 8-iso-PGF(2alpha) production, both basally and after hydrogen peroxide-induced oxidative stimulus. In aged rats, 8-iso-PGF(2alpha) production was not affected by the garlic extract in the basal state, whereas, after hydrogen peroxide-induced oxidative stimulus, an antioxidant effect of the garlic extract appeared only at the higher concentration tested. In conclusion, garlic supplementation could be effective in preventing brain oxidative damage in young animals, whereas the aging brain seems to be resistant to the antioxidant effects of garlic, in vitro.

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.

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 H2O2‐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

Protective Effects Induced by Microwave-Assisted Aqueous Harpagophytum Extract on Rat Cortex Synaptosomes Challenged with Amyloid β-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 β‐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 β‐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 Alzheimers disease. Copyright

Antimicrobial Activity against Beneficial Microorganisms and Chemical Composition of Essential Oil of Mentha suaveolens ssp. insularis Grown in Sardinia

UNLABELLED The aim of this work was to determine the chemical constituents and in vitro antimicrobial activity of the essential oil (EO) of the aerial parts of Mentha sueveolens spp. insularis grown in Sardinia (Italy) against probiotic and starter microorganisms. The gas chromatography-mass spectrometry (GC-MS) analysis allowed to identified 34 compounds, most of oxygenated monoterpene compounds (82.5%) and among them, pulegone was found as major compound (46.5%). The agar diffusion test carried out employing the EO of Mentha suaveolens spp. insularis showed a low antibacterial activity, in particular no action was noticed for probiotic bacteria belonging to lactic acid bacteria groups, whereas almost all yeasts strains tested were inhibited. The automated microtitter dilution assay showed a clear effect at increasing concentration of EO on the specific growth rate (μ) and extension of the lag phase (λ) only for S. xylosus SA23 among bacteria and for Saccharomyces cerevisiae, Tetrapisispora phaffii CBS 4417, Metschnikowia pulcherrima, and Candida zemplinina among yeasts. Results obtained in this work allow us to broaden the knowledge on the effect of EOs on probiotic and food-related microorganisms. PRACTICAL APPLICATION Mentha suaveolens spp. insularis may be used in combination with probiotic bacteria into the food matrix or encapsulated in coating and edible films for food preservation.

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.