Utvrđivanje fenolnih substanci kod šest različitih populacija medvjeđe lijeske (Corylus colurna L.) i usporedba fluktuacije fenola u slučaju nedostatka vode

Turkish hazel (Corylus colurna L.) is one of the naturally grown hazelnut species in Turkey. It can be easily separated from other hazel species with its thick single stem and tall appearance. It can be used in afforestation and erosion control studies due to low demand for habitat and strong root system. It contains substances with high medicinal value in its fruits and leaves. Hazel leaves have strong antioxidant activity due to their high phenolic content. Objective of the study was primarily to determine the individual phenolic constituents of six different population of Turkish Hazelnut and then to evaluate the effect of water deficiency stress generated by irrigation regime on phenolic constituents and photosystem II activity on these genotypes. Grafted plants were produced by taking scions from six different populations of Turkish Hazelnut (Oğuzlar, Erenler, Merkeşler, Seben, Güney Felakettin and Pelitcik). The study was started when the grafted seedlings were 7 years old in greenhouse. During the experiment (June and July), two different levels of irrigation were applied (W1: the soil was fully irrigated to reach field capacity in each irrigation; W2: 50% reduction of W1 irrigation water). After the application of two different irrigation regimes, leaves were collected for each month, dried, extracted with methanol and then quantitatively analyzed and compared for individual phenolic constituents (gallic acid monohydrate, caffeic acid, rutin hydrate, luteolin-7-O-β-D glucoside, kaempferol, rosmarinic acid, myricetin, quercetin, coumarin and apigenin) by using high performance liquid chromatography (HPLC) coupled with a diode array detector (DAD). Generally, rutin, kaempferol and luteolin were dominant individual phenols in methanol extracts of Turkish hazelnut leaves. Pelitcik population was noticeable source of rutin and kaempferol in June, and the halved irrigation regime significantly increased the levels of both phenols in July. Similarly, the highest total phenolic content was observed in the Pelitcik population in June and the halved irrigation regime significantly increased the total phenolic content in both months in this population. It was also determined to what extent water deficiency physiologicallyaffects the quantum efficiency (Fv/Fm) of photosystem II activity through chlorophyll fluorescence technique in hazelnut leaves. Generally, Fv/Fm value decreased with water deficiency. This study showed that water deficiency stress generally caused an increase in phenolic constituents in Turkish Hazel leaves and they may be proper natural sources of phenolic constituents with abiotic stress applications in pharmaceutical and food industry.Medvjeđa lijeska (Corylus colurna L.) jedna je od prirodno rasprostanjenih vrsta lijeske u Turskoj. Lako se može razlikovati od drugih vrsta zahvaljujući svojoj visini i promjeru debla. Vrlo često se koristi za pošumljevanje terena sklonih eroziji zbog snažnog korijenja i skromnih ekloških zahtjeva. Njezino lišće i plodovi sadrže mnogo tvari koje se koriste u medicinske svrhe. Lišće medvjeđe ljeske ima snažno antioksidativno djelovanje zbog visokog sadržaja fenola. Cilj ovoga istraživanja bio je utvrditi sadržaj fenola u lišću šest populacija medvjeđe lijeske te procijeniti učinak sušnoga stresa na sadržaj fenola i kvantnu učinkovitost fotosustava II. Istraživane biljke proizvedene su cijepljenjem nakon sakupljanja plemki iz šest populacija medvjeđe lijeske (Oğuzlar, Erenler, Merkeşler, Seben, Güney Felakettin i Pelitcik). Istraživanje je provedeno u stakleničkim uvjetima kad su kalemljene biljke bile stare 7 godina. Tijekom pokusa (lipanj i srpanj), primijenjena su dva različita načina navodnjavanja (W1: tlo je navodnjavano do poljskog vodnog kapaciteta; W2: tlo je navodnjavano s 50 % manje vode u odnosu na W1 način navodnjavanja). Nakon primjene dva različita režima navodnjavanja, lišće je uzorkovano u lipnju i srpnju. Nakon toga je osušeno, ekstrahirano s metanolom te kvantitativno analizirano s ciljem utvrđivanja njegova fenolnog sastava (galna kiselina monohidrat, kofeinska kiselina, rutin, hidrat, luteolin-7-O-β-D glukozid, kaempferol, rozmarinska kiselina, miricetin, kvercetin, kumarin i apigenin) uz korištenje tekuće kromatografije visoke djelotvornosti (HPLC) i detektora s nizom dioda (DAD). Rutin, kaempferol i luteolin bili su dominantni fenoli u ekstraktu metanola iz lišća medvjeđe lijeske. Populacija Pelitcik bila je značajan izvor rutina i kaempferola u lipnju, a W2 režim navodnjavanja značajno je povećao razine oba fenola u srpnju. Isto tako, najviši sadržaj fenola zabilježen je u populaciji Pelitcik u lipnju, a W2 režim navodnjavanja značajno je povećao ukupan sadržaj fenola u lipnju i srpnju. Također je utvrđeno u kojoj mjeri nedostatak vode utječe na kvantnu učinkovitost fotosustava II (Fv/Fm). Općenito, Fv/Fm vrijednosti opadaju s nedostatkom vode. Ova studija pokazala je da nedostatak vode kod medvjeđe ljeske potiče produkciju fenola te da se na taj način može osigurati prirodni izvor fenola koji se može koristiti u farmaceutskoj i prehrambenoj industriji

Cytotoxic, anti-inflammatory and antioxidant activities of four different extracts of Galega officinalis L (Goat’s rue)

Purpose: To evaluate the cytotoxic, anti-inflammatory and antioxidant activities of four different solvent extracts obtained from the aerial parts of Galega officinalis L.Methods: The hexane, DCM, methanol and water extracts of G. officinalis were successively obtained by soxhlet extraction method. The cytotoxic activity of the extracts was assessed against human lung carcinoma (A-549), human colorectal adenocarcinoma (HT-29), human brain glioblastoma (U-87), and colon adenocarcinoma (DLD-1) by Resazurine test. The antioxidant activity of extracts were determined by Folin-Ciocalteau, oxygen radical absorbing capacity (ORAC), and 2’.7’-dichlorofluorescin-diacetate (DCFH-DA) cell-based assay while their anti-inflammatory activity was determined by nitric oxide (NO) assay.Results: DCM extract showed strong cytotoxic activity against lung adenocarcinoma and brain glioblastoma cell lines, with IC50 (concentration inhibiting 50 % of cell growth) values of 11 ± 0.4 and 16 ± 3 μg/mL, respectively. The hexane extract showed moderate anticancer activity against the same cell lines (59 ± 13 and 63 ± 16 μg/mL, respectively). DCM extract also showed significant anti-inflammatory activity, inhibiting NO release by 86.7 % at 40 μg/mL in lipopolysaccharide (LPS) - stimulated murine RAW 264.7 macrophages. Of all test extracts, the methanol extract of G. officinalis showed the highest antioxidant activity with 2.33 ± 0.09 μmol Trolox/mg , 7.10 ± 0.9 g tannic acid equivalent (TAE), and IC50 of 44 ± 4 μg/mL.Conclusion: The findings of this study suggest that DCM extract may possess anticancer effect against lung adenocarcinoma and brain glioblastoma, as well as serve as an anti-inflammatory agent.Keywords: Galega officinalis L, Biological activity, Bioassay, Anticancer, Lung adenocarcinoma, Brain glioblastoma, Goat’s ru

EVALUATION OF ANTIBACTERIAL, ANTITUMOR, ANTIOXIDANT ACTIVITIES AND PHENOLIC CONSTITUENTS OF FIELD-GROWN AND IN VITRO-GROWN LYSIMACHIA VULGARIS L.

Background: Lysimachia vulgaris L. (Yellow loosestrife) is a medicinal plant in the family Myrsinaceae. It has been used in the treatment of fever, ulcer, diarrhea and wounds in folk medicine. It has also analgesic, expectorant, astringent and antiinflammatory activities. Two different sources of the plant (field-grown and in vitro-grown) were used to evaluate the biological activities (antibacterial, antitumor and antioxidant) of L. vulgaris. In vitro-grown plant materials were collected from L. vulgaris plants that were previously regenerated in our laboratory. Materials and Methods: Plant materials were extracted with water, ethanol and acetone. For antibacterial test, disc diffusion method and 10 different pathogenic bacteria were used. Antioxidant activity was indicated by using DPPH method. The total phenol amount by using Folin-Ciocaltaeu method and the total flavonoid amount by using aluminum chloride (AlCl3) colorimetric method were determined. Results: Generally, yellow loosestrife extracts demonstrated antibacterial activity against Gram-positive bacteria (Staphylococcus aureus, S. epidermidis and Streptococcus pyogenes). Strong antitumor activity of yellow loosestrife was observed via potato disc diffusion bioassay. Nine different phenolics were also determined and compared by using High-Performance Liquid Chromatography (HPLC). Conclusion: Future investigations should be focused on fractionation of the extracts to identify active components for biological activity

Antiproliferative Activity of Some Medicinal Plants on Human Breast and Hepatocellular Carcinoma Cell Lines and their Phenolic Contents

Purpose: To determine the phenolic composition and antiproliferative activity of 16 different extracts (hexane, dichloromethane, methanol and water) obtained from Bellis perennis, Convolvulus galaticus, Trifolium pannonicum and Lysimachia vulgaris on human breast cancer (MCF-7) and human hepatocellular carcinoma (HepG2/C3A) cell lines.Methods: The aerial parts of the plants were successively extracted with hexane, dichloromethane, methanol and water using a Soxhlet apparatus. The phenolic content of the plants were determined by plants by high performance liquid chromatography (HPLC) while their antiproliferative activity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide, a yellow tetrazole (MTT) assay.Results: Among the tested extracts, the methanol extract of B. perennis showed the best antiproliferative activity against MCF-7 cell line with IC50 (inhibiting 50 % of cell growth) value of 71.6 μg/mL. Furthermore, the dichloromethane extract of C. galaticus showed the best anti-proliferative activity against HepG2/C3A cell line with IC50 of 57.3 μg/mL. The HPLC data for the plant extracts showed the presence of the following phenolic compounds: gallic acid monohydrate, caffeic acid, rutin hydrate, luteolin-7-O-β-D glucoside, kaempferol, myricetin, quercetin, coumarin and apigenin.Conclusion: The findings of this study indicate that there is some justification for the use of B. perennis and C. galaticus as traditional anticancer medicinal herbs.Keywords: Bellis perennis, Convolvulus galaticus, Trifolium pannonicum subsp. elongatum, Lysimachia vulgaris, MCF-7, HepG2/C3A, Phenolics, Breast cancer, Antiproliferativ

THE EFFECTS OF OXIDATIVE STRESS ON PHENOLIC COMPOSITION AND ANTIOXIDANT METABOLISM IN CALLUS CULTURE OF COMMON DAISY

Background: Exogenous pretreatment of oxidative stress on callus cultures of Bellis perennis L. (common daisy) induced catalase (CAT), superoxide dismutase (SOD), total phenolic, total flavonoid, total protein and selected commercial phenolic compounds production and accumulation. Materials and Methods: The callus culture obtained from B. perennis pedicel explants was incubated on Murashige and Skoog medium (MS) containing 10 mM H2O2 or 0 mM H2O2 (for control group) for 10 hours. Twenty phenolic compounds (apigenin, caffeic acid, p-coumaric acid, gallic acid, genistein, kaempferol, luteolin, myricetin, procyanidin-C1, quercetin, rutin hydrate, vanillic acid, ferulic acid, salicylic acid, sinapic acid, chlorogenic acid, hesperedin, naringenin, rosmarinic acid and isorhamnetin) were detected by LC-ESI-MS/MS analysis in methanolic extracts of 10 mM H2O2 and control treatments. Results: A predominant phenolic compound was chlorogenic acid followed by rutin hydrate, caffeic acid, luteoline, isorhamnetin, quercetin, myricetin, apigenin, p-coumaric acid and kaempferol. No gallic acid, genistein, procyanidin-C1, vanillic acid, sinapic acid, hesperidin and naringenin were detected in H2O2 treatment and control groups of B. perennis. The total phenolic contents estimated were in the order of H2O2 treatment (285.36 μg/g dw) and control (220.79 μg/g dw) groups. The biosynthesis and accumulation of kaempferol, myricetin, quercetin and isorhamnetin were only determined in H2O2 treatment callus materials. The H2O2 pretreatment clearly showed in a raise in enzymatic and non-enzymatic antioxidant activities. Finally, a significant positive correlation between phenolic accumulation and comprehensive activities of CAT, SOD, total phenolic, total flavonoid and proline was accessible. Conclusion: The present results suggest that using H2O2 as an elicitor or a stimulant plays a significant enhancement role in special phenolic molecules biosynthesis and activation of antioxidant metabolism on callus cultures of B. perennis

Evaluation of Antibacterial and Antitumor Activities of Some Turkish Endemic Plants

Purpose: To investigate the antibacterial and antitumor activities of the aerial parts of 8 different Turkish endemic plants ( Phlomis russeliana , Phlomis armeniaca , Astragalus brachypterus , Astrantia maxima , Ptilostemon afer , Senecio castagneanus , Echium orientale and Arum euxinum ). Methods: Two different bioassays were performed to evaluate the antibacterial and antitumor activities of the endemic plants. For each plant, 3 types of extracts (aqueous, methanol and ethanol) were prepared, giving a total of 24 extracts tested. The disc diffusion assay was used to screen for antibacterial activity against 10 bacteria including Streptococcus pyogenes , Staphylococcus aureus , Staphylococcus epidermidis , Escheria coli , Pseudomonas aeruginosa , Salmonella typhimurium , Serratia marcescens , Proteus vulgaris , Enterobacter cloacea , and Klebsiella pneumoniae . Five different antibiotics (ampicillin, carbenicillin, chloramphenicol, erythromycin and tetracycline) were used as positive controls. Antitumor activity was evaluated using potato disc diffusion bioassay with camptothecin as positive control. Results: The highest antibacterial activity was observed for all extracts of A. brachypterus against S. pyogenes (15.0 - 16.3 mm inhibition zone). The aqueous extract of S. castagneanus showed the strongest antibacterial activity against S. pyogenes (14.3 mm). Furthermore, alcohol extracts (ethanol and methanol) of P. russeliana exhibited moderate activity against S. epidermidis (9 and 8.5 mm, respectively) and S. pyogenes (9.5 mm). High antitumor activity was observed for all extracts of A. brachypterus (91.7 - 100 % tumor inhibition). In addition, ethanol extract of P. russeliana (75 % inhibition) exhibited strong antitumor activity. Conclusion: The present study reveals the strong antibacterial and antitumor activities of A. brachypterus. However, the active components of the plant extracts needs to be identified in future studies

<em>In vitro</em> antibacterial and antitumor efficiency of some traditional plants from Turkey

50-58Traditional usages of medicinal plants can be justified scientifically with bioassays. Three different extracts (aqueous, methanol and ethanol) of 11 Turkish traditional plants [Mentha longifolia (L.) L., Lamium album subsp. crinitum (Montbret & Aucher ex Benth.) Mennema, Sideritis taurica Steph. ex Willd., Hypericum perforatum L., Hypericum linarioides Bosse, Cyanus triumfettii (All.) Dostál ex Á. Löve & D. Löve, Daphne oleoides Schreb., Anemone nemorosa L., Polygonatum orientale Desf., Cynoglossum montanum L. and Hyoscyamus niger L.] were evaluated for their antibacterial and antitumor potentials. Antibacterial activity was evaluated with 3 Gram-positive (Staphylococcus aureus, Staphylococcus epidermidis and Streptococcus pyogenes) and 7 Gram-negative (Serratia marcescens, Salmonella typhimurium, Pseudomonas aeruginosa, Proteus vulgaris, Klebsiella pneumonia, Enterobacter cloacae and Escheria coli) bacteria by using disc diffusion method. Ethanol and methanol extracts of S. taurica, and all extracts of H. perforatum and H. linarioides showed the best antibacterial activities against tested Gram-positive bacteria. Aqueous extract of L. album displayed the best inhibition against Streptococcus pyogenes. Antitumour potential was revealed with Agrobacterium tumefaciens-induced potato disc tumor assay. Strong antitumor activity was observed with ethanol and methanol extracts of L. album (90 % and 80 %, respectively). This study ascertained the scientific rationale behind the traditional knowledge of S. taurica, H. perforatum, H. linarioides, L. album and D. oleoides. Identification of active components should be investigated for plant extracts having strong antibacterial and antitumor activities in further studies.</em

<em>In vitro</em> adventitious plant regeneration of <em>Echium orientale</em> L., an endemic plant: The evaluation of biological activities and phenolic content

264-272Echium orientale L. is a Turkish endemic plant in the family Boraginaceae. The aim of this study was to develop an efficient in vitro culture protocol for E. orientale and compare the field-grown and in vitro-grown leaves in terms of antibacterial, antitumor and antioxidant activities and phenolic profiles. Leaf and petiole explants were cultured on Murashige & Skoog (MS) medium with various plant growth regulator combinations. Best shoot proliferation was obtained from leaf explant with 1 mg/L thidiazuron (TDZ) and 0.5 mg/L indole-3-acetic acid (IAA), producing 8.7 shoots per explant at 100% shoot frequency. Regenerated shoots were transferred to rooting media containing different levels of IAA, indole-3-butyric acid (IBA), naphthalene acetic acid (NAA) or 2,4 dichlorophenoxyacetic acid (2,4-D). The best root formation was observed with MS basal medium (control). Rooted explants were transferred to vermiculate in Magenta containers for acclimatization and after 3 weeks they were transferred to plastic pots containing potting soil and maintained in the plant growth room. Leaves from field-grown plants showed higher antibacterial, antitumor and antioxidant activities, and phenolic content than those from in vitro-grown plants

Cytotoxic, anti-inflammatory and antioxidant activities of four different extracts of <i>Galega officinalis</i> L (Goat’s rue)

Purpose: To evaluate the cytotoxic, anti-inflammatory and antioxidant activities of four different solvent extracts obtained from the aerial parts of Galega officinalis L.Methods: The hexane, DCM, methanol and water extracts of G. officinalis were successively obtained by soxhlet extraction method. The cytotoxic activity of the extracts was assessed against human lung carcinoma (A-549), human colorectal adenocarcinoma (HT-29), human brain glioblastoma (U-87), and colon adenocarcinoma (DLD-1) by Resazurine test. The antioxidant activity of extracts were determined by Folin-Ciocalteau, oxygen radical absorbing capacity (ORAC), and 2’.7’-dichlorofluorescin-diacetate (DCFH-DA) cell-based assay while their anti-inflammatory activity was determined by nitric oxide (NO) assay.Results: DCM extract showed strong cytotoxic activity against lung adenocarcinoma and brain glioblastoma cell lines, with IC50 (concentration inhibiting 50 % of cell growth) values of 11 ± 0.4 and 16 ± 3 μg/mL, respectively. The hexane extract showed moderate anticancer activity against the same cell lines (59 ± 13 and 63 ± 16 μg/mL, respectively). DCM extract also showed significant anti-inflammatory activity, inhibiting NO release by 86.7 % at 40 μg/mL in lipopolysaccharide (LPS) - stimulated murine RAW 264.7 macrophages. Of all test extracts, the methanol extract of G. officinalis showed the highest antioxidant activity with 2.33 ± 0.09 μmol Trolox/mg , 7.10 ± 0.9 g tannic acid equivalent (TAE), and IC50 of 44 ± 4 μg/mL.Conclusion: The findings of this study suggest that DCM extract may possess anticancer effect against lung adenocarcinoma and brain glioblastoma, as well as serve as an anti-inflammatory agent.Keywords: Galega officinalis L, Biological activity, Bioassay, Anticancer, Lung adenocarcinoma, Brain glioblastoma, Goat’s ru