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Öğe Analytical Procedures for Secondary Metabolites Determination: Recent Trends and Future Perspectives(BENTHAM SCIENCE PUBL LTD, 2018) Menghini, Luigi; Bellagamba, Giuseppe; Fabiana, Marcelli; De Luca, Elisa; Armellani, Luca; Cifelli, Roberta; Granese, AriannaBackground: Many studies have shown the benefits deriving from plants secondary metabolites to human health also for the prevention of several diseases associated to oxidative stress including cancer, cardiovascular diseases and diabetes mellitus. Nowadays there is a high attention on plant secondary metabolites and analytical techniques related to their identification and characterization. Conclusion: This review aims to report analytical procedures, instrumental configurations, and extraction approaches devoted to secondary metabolites analyses. In this direction, an introduction to secondary metabolites is made with emphasis on their formation, biological and pharmaceutical roles. Extraction, characterization and analytical procedures are described in the second part of this review, while a comparison of the analytical techniques will be discussed in the third part. This review could provide a better understanding of the analytical procedure and techniques for secondary metabolites and their applications in the scientific and industrial areas including food, food supplements, cosmetic, and pharmaceutical, especially related to quality control analyses and to chemical fingerprint elucidation, all in order to standardize these plant-derived extracts before in vitro and in vivo studies.Öğe Chromatographic analyses, in vitro biological activities, and cytotoxicity of cannabis sativa l. essential oil: a multidisciplinary study(MDPI, 2018) Zengin, Gökhan; Menghini, Luigi; Sotto, Antonella Di; Mancinelli, Romina; Sisto, Francesca; Carradori, Simone; Cesa, Stefania; Fraschetti, Caterina; Filippi, Antonello; Angiolella, Letizia; Locatelli, Marcello; Mannina, Luisa; Ingallina, Cinzia; Puca, Valentina; D’Antonio, Marianna; Grande, RossellaDue to renewed interest in the cultivation and production of Italian Cannabis sativa L., we proposed a multi-methodological approach to explore chemically and biologically both the essential oil and the aromatic water of this plant. We reported the chemical composition in terms of cannabinoid content, volatile component, phenolic and flavonoid pattern, and color characteristics. Then, we demonstrated the ethnopharmacological relevance of this plant cultivated in Italy as a source of antioxidant compounds toward a large panel of enzymes (pancreatic lipase, -amylase, -glucosidase, and cholinesterases) and selected clinically relevant, multidrug-sensible, and multidrug-resistant microbial strains (Staphylococcus aureus, Helicobacter pylori, Candida, and Malassezia spp.), evaluating the cytotoxic effects against normal and malignant cell lines. Preliminary in vivo cytotoxicity was also performed on Galleria mellonella larvae. The results corroborate the use of this natural product as a rich source of important biologically active molecules with particular emphasis on the role exerted by naringenin, one of the most important secondary metabolites.Öğe A comparative assessment of biological effects and chemical profile of Italian asphodeline lutea extracts(MDPI, 2018) Melucci, Dora; Locatelli, Marcello; Locatelli, Clinio; Zappi, Alessandro; De laurentiis, Francesco; Carradori, Simone; Campestre, Cristina; Leporini, Lidia; Zengin, Gökhan; Picot, Carene Marie Nancy; Menghini, Luigi; Mahomoodally, Mohamad FawziThe 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 alpha-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.Öğe Comparative study of biological activities and multicomponent pattern of two wild Turkish species: Asphodeline anatolica and Potentilla speciosa(TAYLOR & FRANCIS LTD, 2016) Zengin, Gökhan; Menghini, Luigi; Malatesta, Luciano; De Luca, Elisa; Bellagamba, Giuseppe; Uysal, Şengül; Aktümsek, AbdurrahmanThe 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.Öğe Comprehensive Chemical Profiling and Multidirectional Biological Investigation of Two Wild Anthemis Species (Anthemis tinctoria var. Pallida and A. cretica subsp. tenuiloba): Focus on Neuroprotective Effects(MDPI, 2019) Orlando, Giustino; Zengin, Gökhan; Ferrante, Claudio; Ronci, Maurizio; Recinella, Lucia; Şenkardeş, İsmail; Gevrenova, Reneta; Zheleva-Dimitrova, Dimitrina; Chiavaroli, Annalisa; Leone, Sheila; Di Simone, Simonetta; Brunetti, Luigi; Picot-Allain, Carene Marie Nancy; Mahomoodally, Mohamad Fawzi; Sinan, Kouadio Ibrahime; Menghini, LuigiEthyl acetate (EA), methanol (MeOH), and aqueous extracts of aerial parts of Anthemis tinctoria var. pallida (ATP) and A. cretica subsp. tenuiloba (ACT) were investigated for their phenol and flavonoid content, antioxidant, and key enzyme inhibitory potentials. All extracts displayed antiradical effects, with MeOH and aqueous extracts being a superior source of antioxidants. On the other hand, EA and MeOH extracts were potent against AChE and BChE. Enzyme inhibitory effects against tyrosinase and alpha-glucosidase were observed, as well. We also studied Anthemis extracts in an ex vivo experimental neurotoxicity paradigm. We assayed extract influence on oxidative stress and neurotransmission biomarkers, including lactate dehydrogenase (LDH) and serotonin (5-HT), in isolated rat cortex challenged with K+ 60 mM Krebs-Ringer buffer (excitotoxicity stimulus). An untargeted proteomic analysis was finally performed in order to explore the putative mechanism in the brain. The pharmacological study highlighted the capability of ACT water extract to blunt K+ 60 mM increase in LDH level and 5-HT turnover, and restore physiological activity of specific proteins involved in neuron morphology and neurotransmission, including NEFMs, VAMP-2, and PKC gamma, thus further supporting the neuroprotective role of ACT water extract.Öğe Crocus Sativus, Serenoa Repens and Pinus Massoniana Extracts Modulate Inflammatory Response in Isolated Rat Prostate Challenged with LPS(BIOLIFE SAS, 2017) Chiavaroli, Annalisa; Recinella, Lucia; Ferrante, Claudio; Locatelli, Marcello; Carradori, Simone; Macchione, Nicola; Zengin, Gökhan; Leporini, Lidia; Leone, Sheila; Martinotti, S.; Brunetti, L.; Vacca, M.; Menghini, Luigi; Orlando, GiuliaProstatitis is a common prostate disease that could be promoted by bacterial or non-bacterial infectious agents. In addition, inflammatory pathways involved in prostatitis have been increasingly studied, and herbal extracts endowed with anti-inflammatory effects are under investigation, individually or in combination, for their efficacy in alleviating the burden of inflammation, with possible improvements in symptoms. Serenoa repens (Serenoa), in combination with Crocus sativus (Crocus) and Pinus massoniana (Pinus), has previously shown to improve sexual function and limit urinary symptoms in patients suffering from concomitant erectile dysfunction and lower urinary tract symptoms. In this context, the aim of the present study is to evaluate the efficacy of Serenoa, Crocus and Pinus extracts, either alone or in combination, on immortalized prostate cells (PC3) and in an experimental model of bacterial prostatitis constituted by ex vivo prostate specimens challenged with lipopolysaccharide (LPS). We found that the tested extracts were able to reduce ROS production by PC3 cells and NF kappa B and PGE(2) activity in prostate specimens challenged with LPS. In addition, the pharmacological association of the extracts displayed synergistic effects indicating a rational use of the mixture of the tested extracts as a novel anti-oxidant and anti-inflammatory formulation in bacterial prostatitis. Finally, we performed analytical and in vitro evaluation to better characterize the phytochemical profile and the mechanism of action of selected secondary metabolites.Öğe Multidirectional biological investigation and phytochemical profile of Rubus sanctus and Rubus ibericus(PERGAMON-ELSEVIER SCIENCE LTD, 2019) Zengin, Gökhan; Ferrante, Claudio; Senkardes, Ismail; Gevrenova, Reneta; Zheleva-Dimitrova, Dimitrina; Menghini, Luigi; Orlando, Giustino; Recinella, Lucia; Chiavaroli, Annalisa; Leone, Sheila; Brunetti, Luigi; Picot-Allain, Carene Marie Nancy; Rengasamy, Kannan RR.; Mahomoodally, Mohamad FawziIn the present study, the biological properties, including, the enzyme inhibitory and antioxidant activities, as well as, the phytochemical profile of the ethyl acetate, methanol, and water extracts of Rubus sanctus Schreb. and Rubus ibericus Juz. leaves were determined using in vitro bioassays. Wide range of phytochemicals, including, hydroxybenzoic acids, hydroxycinnamic acids, acylquinic acids, ellagitannins, flavonoids, and triterpenoid saponins were determined using UHPLC-ESI/HRMS technique. The ethyl acetate and methanol extracts of the studied Rubus species effectively inhibited acetyl and butyryl cholinesterase. On the other hand, R. sanctus water extract showed low inhibition against alpha-amylase and prominent inhibitory action against alpha-glucosidase. Data collected from this study reported the radical scavenging and reducing potential of the studied Rubus species. Investigation of the protective effects of the different extracts of R. sanctus and R. ibericus in experimental model of ulcerative colitis was performed. The extracts were also tested on spontaneous migration of human colon cancer cells (HCT116) in wound healing experimental paradigm. Only R. sanctus methanol extract inhibited spontaneous HCT116 migration in the wound healing test. Our results suggested that R. sanctus and R. ibericus may be potential candidates as sources of biologically-active compounds for the development of nutraceuticals, pharmaceuticals, and/or cosmetics.Öğe Multiple pharmacological approaches on hydroalcoholic extracts from different parts of Cynoglossum creticum Mill. (Boraginaceae)(TAYLOR & FRANCIS LTD, 2019) Menghini, Luigi; Ferrante, Claudio; Zengin, Gökhan; Mahomoodally, Mohamad Fawzi; Leporini, Lidia; Locatelli, Marcello; Cacciagrano, Francesco; Recinella, Lucia; Chiavaroli, Annalisa; Leone, Sheila; Brunetti, Luigi; Orlando, GiustinoCynoglossum creticum Mill (Boraginaceae) is used traditionally as a remedy to manage several human ailments. In this context, the present study aimed to perform multiple pharmacological investigations on the hydroalcoholic extracts prepared from Cynoglossum roots and aerial parts (leaves and flowers). We evaluated the antioxidant and enzyme inhibitory (against cholinesterases, alpha-glucosidase, alpha-amylase, lipase and tyrosinase) activity of the extracts. The protective effect(s) of the extracts on cardiomyocyte C2C12 and intestinal HCT116 cell lines challenged with hydrogen peroxide (H2O2) was studied. We found that the aerial parts harbored the highest amount of phenolic compounds. Generally, aerial parts showed significant antioxidant and enzyme inhibitory effects. Leaves exhibited the best lipase inhibitory activity (173.15 mgOE/g extract), followed by flowers and roots. The root and aerial extracts were equally able to blunt intracellular H2O2 induced reactive oxygen species production from both C2C12 and HCT116 cell lines. Both cells lines could be treated with scalar concentrations of root and flower extracts in the range 50-300 mu g/mL without interferences on cell viability. In conclusion, the present study showed protective effects exerted by Cynoglossum extracts, which could serve as a foundation for the development of pharmaceuticals and nutraceuticals derived from Cynoglossum.Öğe New pharmacological targets of three Asphodeline species using in vitro and ex vivo models of inflammation and oxidative stress(TAYLOR & FRANCIS LTD, 2019) Zengin, Gökhan; Locatelli, Marcello; Ferrante, Claudio; Menghini, Luigi; Orlando, Giustino; Brunetti, Luigi; Recinella, Lucia; Chiavaroli, Annalisa; Leone, Sheila; Leporini, Lidia; Aumeeruddy, Muhammad Zakariyyah; Mahomoodally, Mohamad FawziThis 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 F2 alpha (8-iso-PGF2 alpha) levels, 5HIAA/5-HT ratio, tumor necrosis factor (TNF)-alpha 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-alpha 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.Öğe Nutraceutical potential of Corylus avellana daily supplements for obesity and related dysmetabolism(ELSEVIER SCIENCE BV, 2018) Mollica, Adriano; Zengin, Gökhan; Stefanucci, Azzurra; Ferrante, Claudio; Menghini, Luigi; Orlando, Giustino; Brunetti, LuigiIn this study, the nutraceutical potential of two hazelnut varieties (Turkey and Italy) were examined by studying several aspects including the phytochemical properties, antioxidant potential, cell viability and ex vivo neuroprotective potential. Results showed that the Turkish hazelnut had the higher concentration of phenolic acids, flavonoids, higher antioxidant capacity and enzyme inhibition properties and lower saturated fatty acid concentration than the Italian sample. The in vivo studies showed that compared to the Italian hazelnuts, the addition of Turkish hazelnuts to high fat diet was associated with a more significant decrease in body weight, food consumption, atherogenic index, lipid peroxidation levels and biochemical/morphological markers of liver injury. Also, from our results, the two hazelnut varieties were protective against beta-amyloid-induced neurochemical changes and high-fat diet induced alteration of metabolic indices.Öğe Phytochemical and Pharmacological Profile of Origanum Sipyleum Extracts: Exploring Novel Sources for Potential Pharmaceutical, Food, and Cosmetic Applications(Georg Thieme Verlag KG, 2019) Ferrante, Claudio; Zengin, Gökhan; Orlando, Giulia; Recinella, Lucia; Chiavaroli, Annalisa; Leone, Silvia; Brunetti, L.; Menghini, LuigiOriganum sipyleum L., an endemic plant of Western Anatolia has been used as a medicinal tea, food additive, and for the production of essential oil. In this study, the biological potential of three extracts (ethyl acetate, methanol, and aqueous) of O. sipyleum was assessed based on antioxidant activity against key enzymes of clinical relvance. The chemical profile of the plant was assessed using spectrophotometric and LC-MS techniques. Additionally, we explored potential antioxidant and anti-inflammatory effects duced by the extracts in an experimental model of ulcerative colitis induced by LPS challenging. LC-MS analysis revealed that the extracts contained different classes of phenolics, such as rosmarinic acid, phlorizin and gallic acid. We found that the aqueous extract was the most effective antioxidant, displaying the highest DPPH and ABTS scavenging, FRAP, CUPRAC, molybdenum(VI) reducing, and metal chelating effect. The aqueous extract howed the strongest acetylcholinesterase (AChE) inhibition; the methanol extract showed the highest ?-glucosidase inhibition, while the ethyl acetate extract was the most effective on butyrylcholinesterase (BChE), tyrosinase, and ?-amylase. The total flavonoid content was highest in the aqueous and ethyl acetate extract, respectively. Finally, we found that all extracts were effective in reducing LPS-induced activity of pro-oxidant and pro-inflammatory biomarkers including nitrites, LDH, PGE2 and 5-HT, in rat colon, with the best activity showed by ethyl acetate extract. Our results indicated that the three solvent extracts varied in their chemical and biological profiles, but overall, O. sipyleum showed promising therapeutic properties, nonetheless, need to be further validated in in vivo models.Öğe Protective effects of Cotoneaster integerrimus on in vitro and ex-vivo models of H2O2-induced lactate dehydrogenase activity in HCT116 cell and on lipopolysaccharide-induced inflammation in rat colon(WILEY, 2019) Zengin, Gökhan; Ferrante, Claudio; Menghini, Luigi; Orlando, Giustino; Brunetti, Luigi; Recinella, Lucia; Chiavaroli, Annalisa; Leone, Sheila; Ronci, Maurizio; Aumeeruddy, Muhammad Zakariyyah; Mahomoodally, Mohamad FawziThe present study evaluated the biological potential of methanol and aqueous extracts of the twigs and fruits of Cotoneaster integerrimus Medik. Lethality bioassays performed on Artemia salina showed that aqueous and methanol C. integerrimus extracts were non-toxic in the concentration range (0.1-20 mg/ml), with a LC50 >= 2.5 mg/ml, for each single extract. The protective effect of the extracts was assessed in vitro against hydrogen peroxide-induced lactate dehydrogenase (LDH) activity and tumor necrosis factor (TNF)alpha gene expression in colon cancer HCT116 cell line. All the extracts downregulated (H2O2)-induced TNF alpha gene expression, in HCT116. By contrast, it was observed that the lipopolysaccharide (LPS)-induced increase in colon nitrite, prostaglandin E-2, and 8-iso-PGF(2 alpha) levels were counteracted mostly by the methanol twig extract. The present study showed protective effects induced by C. integerrimus in vitro and ex vivo, thus supporting potential application in the management of chronic inflammatory diseases.Öğe Qualitative Chemical Characterization and Multidirectional Biological Investigation of Leaves and Bark Extracts of Anogeissus leiocarpus (DC.) Guill. & Perr. (Combretaceae)(MDPI, 2019) Orlando, Giustino; Ferrante, Claudio; Zengin, Gökhan; Sinan, Kouadio Ibrahime; Bene, Kouadio; Diuzheva, Alina; Jeko, Jozsef; Cziáky, Zoltán; Di Simone, Simonetta; Recinella, Lucia; Chiavaroli, Annalisa; Leone, Sheila; Brunetti, Luigi; Picot-Allain, Carene Marie Nancy; Mahomoodally, Mohamad Fawzi; Menghini, LuigiAnogeissus leiocarpus (DC.) Guill. & Perr. (Combretaceae) has a long history of use by folk populations for the management of multiple human ailments. Based on the published literature, there has been no attempt to conduct a comparative assessment of the biological activity and the phytochemical profiles of the leaves and stem bark of A. leiocarpus extracted using methanol, ethyl acetate, and water. By high-performance liquid chromatography with electrospray ionization mass spectrometric detection (HPLC-ESI-MSn) analysis, quinic, shikimic, gallic, and protocatechuic acids were tentatively identified from all the extracts, while chlorogenic, caffeic, ferulic, and dodecanedioic acids were only characterised from the leaves extracts. Additionally, a pharmacological study was carried out to evaluate potential protective effects that are induced by the extracts in rat colon and colon cancer HCT116 cell line. In general, the methanol and water extracts of A. leiocarpus leaves and stem bark showed potent radical scavenging and reducing properties. It was noted that the stem bark extracts were more potent antioxidants as compared to the leaves extracts. The methanol extract of A. leiocarpus leaves showed the highest acetyl (4.68 mg galantamine equivalent/g) and butyryl (4.0 mg galantamine equivalent/g) cholinesterase inhibition. Among ethyl acetate extracts, the pharmacological investigation suggested stem bark ethyl acetate extracts to be the most promising. This extract revealed ability to protect rat colon from lipopolysaccharide-induced oxidative stress, without exerting promoting effects on HCT116 cell line viability and migration. As a conclusion, A. leiocarpus represents a potential source of bioactive compounds in the development of novel therapeutic agents.Öğe Qualitative fingerprint analysis and multidirectional assessment of different crude extracts and essential oil from wild Artemisia santonicum L.(MDPI, 2019) Ferrante, Claudio; Zengin, Gökhan; Menghini, Luigi; Diuzheva, Alina; Jeko, Jozsef; Cziaky, Zoltan; Recinella, Lucia; Chiavaroli, Annalisa; Leone, Sheila; Brunetti, Luigi; Lobine, Devina; Şenkardeş, İsmail; Mahomoodally, Mohamad Fawzi; Orlando, GiustinoArtemisia species are used as folk medicines in several countries. This work was aimed to shed more light on the effect of methanol, water, ethyl acetate extracts, and essential oil (EO) of A. santonicum on selected enzymes (cholinesterase, tyrosinase alpha-amylase, and alpha-glucosidase) as well of their antioxidant and pharmacological effects. The chemical profile of the essential oil was determined using gas chromatography coupled to mass spectrometry (GC-MS) analysis, while the extracts were chemically characterized by high performance liquid chromatography coupled to mass spectrometry (HPLC-MS). Forty-nine constituents were identified and camphor (36.6%), 1,8-cineole (10.2%), alpha-thujone (10.1%), borneol (4.5%), and beta-thujone (3.6%) were the major components. Overall, 45, 74, and 67 components were identified from the ethyl acetate, methanol, and water extracts, respectively. The EO and extracts showed significant antioxidant properties, in a cell-free model; particularly, methanol and water extracts revealed promising sources of antioxidant compounds. Additionally, we evaluated protective effects of EO and extracts in isolated rat colon tissue challenged with lipopolysaccharide (LPS), as an ex vivo model of colon inflammation, and human colon cancer HCT116 cell line. Particularly, we observed that, among all tested samples, A. santonicum ethyl acetate displayed the best pharmacological profile, being able to blunt LPS-induced levels of all tested biomarkers of inflammation and oxidative stress, including colon nitrites, lactate dehydrogenase, prostaglandin E-2, and serotonin. Additionally, this extract was also able to reduce HCT116 cell viability, thus suggesting potential antiproliferative effects against colon cancer cells. Based on our results, A. santonicum has great potential for developing novel functional agents including pharmaceuticals, cosmeceuticals, and nutraceuticals.
