DNA protection, molecular docking, antioxidant, antibacterial, enzyme inhibition, and enzyme kinetic studies for parietin, isolated from Xanthoria parietina (L.) Th. Fr.

Parietin was isolated from Xanthoria parietina (L.) Th. Fr.' (methanol:chloroform) extract, using a silica column. 13 C NMR and 1H NMR were used to confirm the structure of the isolated parietin. For the first time, parietin was investigated for its antioxidant, antibacterial and DNA protective activities. Molecular docking was carried out to determine the binding affinity and interactions between the enzymes and our molecule. Inhibition and kinetic mechanism studies for the action of the enzymes were performed too. Parietin exhibited high metal chelating activity. The MIC values of parietin were sufficient to inhibit different bacterial strains; E. coli, P. aeruginosa, K. pneumoniae and S. aureus. Molecular docking applications exhibited that acetylcholinesterase (AChE), butyrylcholinesterase (BChE), lipase, and tyrosinase have high potential for binding with the parietin. Especially, the parietin's highest binding affinity was recorded with AChE and tyrosinase. These results were confirmed by the inhibition and kinetics results, where, parietin observed a potent inhibition with an IC50 values between 0.013-0.003 µM. Moreover, parietin acts' as a non-competitive inhibitor against AChE, BChE, and lipase, and as a competitive inhibitor against tyrosinase with a high rate of inhibition stability. The promising biological properties of parietin revealed its effectiveness in terms of suitability in the food and pharmaceutical industries.Communicated by Ramaswamy H. Sarma.

Two new compounds from endemic Centaurea paphlagonica (Bornm.) Wagenitz and their cytotoxic activities.

Centaurea paphlagonica (Bornm.) Wagenitz is an endemic plant in Turkey. Pyrocatechol, vanillic acid, 3,4-dihydroxy benzoic acid, 5-O-caffeoylshikimic acid, tamarixetin, chlorogenic acid methyl ester, quercetin, 1,3-dicaffeoylquinic acid, tamarixetin-7-O-ß-D-glucopyranoside, quercimetrin, daucosterin, paphlagonicanin B, tamarixetin-7-O-ß-rutinoside, rutin, chlorogenic acid, isoorientin, orientin, 3-O-feruloylquinic acid, quercetagetin-3-methyl ether 6-O-ß-glucopyranoside, diosmetin 6-C-ß-glucopyranoside, quercetagetin 4'-methyl ether 7-O-ß-glucopyranoside, paphlagonicanin A, nepetin, cirsiliol, desacylcynaropicrin, and 8α-O-(2',3'-dihydroxyisobutyryl) desacylcynaropicrin were isolated from both flower and aerial parts of C. paphlagonica. These compounds were identified using 1D and 2D NMR methods and ESI-MS. The MTT assay assessed the antiproliferative activities of all isolated (known and new compounds) compounds on Caco-2, LNCaP, A549, HeLa, and HEK-293 cell lines. The 8α-O-(2',3'-dihydroxyisobutyryl) desacylcynaropicrin demonstrated the highest activity against CaCo-2 and HeLa cancer cell lines.

Determination of antioxidant, DNA protection, enzyme inhibition potential and molecular docking studies of a biomarker ursolic acid in Nepeta species.

Ursolic acid (UA), which has many biological properties such as anti-cancer, anti-inflammatory and antioxidant, and regulates some pharmacological processes, has been isolated from the flowers, leaves, berries and fruits of many plant species. In this work, UA was purified from the methanol-chloroform crude extract of Nepeta species (N. aristata, N. baytopii, N. italica, N. trachonitica, N. stenantha) using a silica gel column with chloroform or ethyl acetate solvents via bioactivity-guided isolation. The most active sub-fractions were determined under bioactivities using antioxidant and DNA protection activities and enzyme inhibitions. UA was purified from these fractions and its structure was elucidated by NMR spectroscopy techniques. The highest amount of UA was found in N. stenantha (8.53 mg UA/g), while the lowest amount of UA was found in N. trachonitica (1.92 mg UA/g). The bioactivities of UA were evaluated with antioxidant and DNA protection activities, enzyme inhibitions, kinetics and interactions. The inhibition values (IC50) of α-amylase, α-glucosidase, urease, CA, tyrosinase, lipase, AChE, and BChE were determined between 5.08 and 181.96 µM. In contrast, Ki values of enzyme inhibition kinetics were observed between 0.04 and 0.20 mM. In addition, Ki values of these enzymes for enzyme-UA interactions were calculated as 0.38, 0.86, 0.45, 1.01, 0.23, 0.41, 0.01 and 2.24 µM, respectively. It is supported that UA can be widely used as a good antioxidant against oxidative damage, an effective DNA protector against genetic diseases, and a suitable inhibitor for metabolizing enzymes.Communicated by Ramaswamy H. Sarma.

The cytotoxicity and antioxidant activity analysis of the isolated constituents and extracts from endemic Centaurea derderiifolia.

It is aimed to investigate the cytotoxic effects of the various extracts from the leaf and seed of Centaurea derderiifolia on the growth human cervical adenocarcinoma (HeLa) cells by xCELLigence method and to isolate the cytotoxic constituents. The results showed that subfractions 2 and 3 exhibited remarkable inhibitory effect against HeLa (IC50 < 10 µg/mL). The chloroform extract of leaf displayed the highest cytotoxic effect on HeLa cells (IC50 < 50 µg/mL) and was therefore subjected to a bioassay-guided multistep separation procedure. The pure compounds were elucidated by spectroscopic and mass-spectrometric analyses, including 1 D-, 2 D-NMR. In addition to cytotoxic effects of the isolated constituents, their antioxidant activities were also studied. On the other hand, subfraction 4 exhibited the highest DPPH radical scavenging activity (IC50 = 0.76 ± 0.03 mg/mL). ß-sitosterol-3-O-ß-d-glucopyranoside was isolated for the first time from this plant and three compounds from the bioactive subfractions were identified.

In vitro antioxidant, DNA-damaged protection and antiproliferative activities of ethyl acetate and n-butanol extracts of Centaurea sphaerocephalaL.

This study aimed to evaluate the in vitro antiproliferative and inhibition of oxidative DNA-damage activities of n-butanol (n-BuOH) extract of Centaurea sphaerocephala. The in vitro antioxidant activity of the ethyl acetate (EtOAc) and the n-BuOH extracts of this plant were also assayed. To investigate the antioxidant potential, extracts were tested for their capacity to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH·) and to inhibit lipid peroxidation using the TBARs method. The contents of total phenolics and flavonoids were measured. Additionally, antiproliferative activity and DNA-damage inhibition of the n-BuOH extract was determined using XCELLigence RTCA instrument and photolyzing 46966 plasmid, respectively. The results exhibited that the scavenging abilities of the EtOAc extract were better than the n-BuOH extract with an IC50= 11.59 µg/mL and 16.67 µg/mL for both extracts, respectively. The phenolic and flavonoid contents were found higher in the n-BuOH and EtOAc extracts. Furthermore, our results showed that n-BuOH extract exhibited a remarkable inhibition of lipid peroxidation with an IC50 of 340.94±7.49 µg/mL and had an antiproliferative effect against Hela cells. Extracts of C. sphaerocephala showed antioxidant activity on scavenging DPPH·. In addition, the n-BuOH extract inhibited the lipid peroxidation and exhibited an antiproliferative effect against HeLa cells line (human cervix carcinoma).

Hepatoprotective effects of the n-butanol extract from Perralderia coronopifolia Coss. against PCP-induced toxicity in Wistar albino rats.

In the present study, in vivo antioxidant properties of the n-butanol extract obtained from aerial parts of Perralderia coronopifolia were investigated in term of its hepatoprotective effect of female Wistar albino rats (n, 36; average age, 48 ± 5 days; weighing 150 ± 18 g) against PCP (pentachlorphenol)-induced toxicity. PCP (20 mg/kg b.w.) and plant extract (50 mg/kg b.w.) were administered daily by gavages for 2 weeks. Vitamin E (100 mg/kg b.w.) was given intraperitoneally as a positive control. Lipid peroxidation (LPO) levels, reduced glutathione (GSH) levels, and glutathione peroxidase (GPx) activities were evaluated in liver homogenates. While, aspartate aminotransferase (AST), alanine aminotransferase (ALT), cholesterol, and triglyceride parameters were analyzed in serums. The liver fragments were observed using light microscopy. Experimental results exhibited that PCP-treated group has a significant increase in the liver lipid peroxidation (LPO) levels of animals while decreased in plant extract-treated group. In addition, PCP caused significant decreases in glutathione peroxidase (GPx) activities and reduced glutathione (GSH) levels. Moreover, PCP induced hepatotoxicity by increasing serum transaminase enzymes, cholesterol, and triglyceride levels. While, these levels were restored to control value in animals treated with plant extract. The regularized levels of LPO, GSH, cholesterol, triglyceride, transaminase enzymes, and GPx activities revealed the antioxidant properties of the extract plant as well as of the vitamin E. The histological study showed the hepatoprotective effect of our extracts against PCP-induced acute intoxication, protecting the hepatic architecture and decreasing the functional and structural alterations of the liver. The plant extract had high antioxidant potential and completely prevented the toxic effect of PCP on the above of liver and serum parameters.

Extraction, isolation of heat-resistance phenolic compounds, antioxidant properties, characterization and purification of 5-hydroxymaltol from Turkish apple pulps.

Apple pulps (AP) were obtained as a side product in fruit juice factories and contains valuable phenolic compounds. The dried AP was subjected to extraction with water, ethyl acetate (APEA) and n-butanol (APBU), respectively. 5-Hydroxymaltol (5-HM) was isolated and confirmed by NMR techniques. The HPLC-TOF/MS analysis revealed the presence of 16 components including major components of morine, gentisic, 4-hydroxybenzoic, vanillic and fumaric acid. The antioxidant activities were evaluated with total antioxidant activity, reducing power, inhibition of lipid peroxidation, metal chelating, free radical and H2O2 scavenging activities. 5-HM, APEA and APBU exhibited the in vitro antioxidant activities in a concentration-dependent and moderate manner. The IC50 values were effective for free radical scavenging activity of 5-HM (8.22 µg mL-1), H2O2 scavenging activity for APEA (8.12 µg mL-1) and inhibition of lipid peroxidation for APEA (0.93 µg mL-1). The 5-HM and APEA have antioxidant capacities and also feasible to apply variety in vivo tests.

Phytochemical Constituents, ChEs and Urease Inhibitions, Antiproliferative and Antioxidant Properties of Elaeagnus umbellata Thunb.

AIM AND OBJECTIVE: Due to the common ethnopharmacological used or scientifically examined biochemical properties, Elaeagnaceae family, Elaeagnus umbellate (Thunb.) (EU, Guz yemisi) was worth investigating. MATERIALS AND METHODS: In this investigation, we revealed antioxidant, antiproliferative and enzyme inhibition activities of the water, methanol, ethanol, acetone, ethyl acetate and hexane extracts of EU as well as the contents of their phenolic, flavonoid, anthocyanin, ascorbic acid, lycopene and ß- carotene. The antioxidant activity was screened by total antioxidant (phosphomolybdenum), inhibition of linoleic acid peroxidation, reducing power, 2-deoxyribose degradation assay, H2O2 scavenging and metal chelating activities of the samples were tested in vitro. Additionally, the scavenging activities of the extracts were determined against 1,1-diphenyl-2-picrylhydrazyl (DPPH˙), 2,2-azino-bis(3-ethylbenzothiazloine-6-sulfonicacid (ABTS˙+), superoxide anion and peroxide radicals. The samples were determined for their inhibitory activities against urease, acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE). In vitro, antiproliferative activities of six different extracts were tested using the xCELLigence system against HeLa and HT29 cell lines. RESULTS: The antioxidant activities of the extracts were found higher than standard antioxidants. The water extracts of fruit and leaf showed the best antioxidant activity. In inhibition assays of urease, AChE and BuChE, all extracts exhibited remarkable inhibition potential. Ethyl acetate extracts, especially, showed better inhibition capacity. It was found that the antioxidant activities of the extracts presented consistently with their chemical contents. The antiproliferative activities of leaf extracts were more effective than the fruit extracts. The chromatographic methods were applied to the different solvents to analyses phenolic secondery metabolites. It was found that fumaric acid, 4- hydroxybenzoic acid, rutin and quercetin-3-ß-D-glucoside, neohesperidin, hesperidin determined to have higher contents all the extracts. CONCLUSION: EU can be suggested as a potential natural source of antioxidants appropriate for utilization in nutritional/pharmaceutical fields.

Protective activity of Hertia cheirifolia extracts against DNA damage, lipid peroxidation and protein oxidation.

CONTEXT: Hertia cheirifolia L. (Asteraceae), a perennial shrub widely distributed in Northern Africa, is traditionally used to treat inflammatory disorders. OBJECTIVE: The protective effect of methanol (Met E) and aqueous (Aq E) extracts of Hertia cheirifolia against DNA, lipid and protein oxidation was investigated. MATERIALS AND METHODS: Different concentrations (50-1000 µg/mL) of Hertia cheirifolia aerial part extracts were examined against DNA, lipid and protein oxidation induced by H2O2 + UV, FeSO4, and Fe3+/H2O2-ascorbic acid, respectively. The DPPH•, metal ion chelating, reducing power and ß-carotene bleaching tests were conducted. RESULTS: Both extracts were rich in polyphenols, flavonoids and tannins, and were able to scavenge DPPH• with IC50 values of 138 and 197 µg/mL, respectively. At 300 µg/mL, Aq E exerted stronger chelating effect (99%) than Met E (69%). However, Met E reducing power (IC50 = 61 µg/mL) was more than that of Aq E (IC50 = 193 µg/mL). Both extracts protected from ß-carotene bleaching by 74% and 94%, respectively, and inhibited linoleic acid peroxidation. The inhibitory activity of Aq E extract (64%) was twice more than that of Met E (32%). Interestingly, both extracts protected DNA against the cleavage by about 96-98%. At 1 mg/mL, Met E and Aq E restored protein band intensity by 94-99%. CONCLUSIONS: Hertia cheirifolia exhibits potent antioxidant activity and protects biomolecules against oxidative damage; hence, it may serve as potential source of natural antioxidant for pharmaceutical applications and food preservation. This is the first report on the protective activity of this plant against biomolecule oxidation.

The impacts of Elaeagnus umbellata Thunb. leaf and fruit aqueous extracts on mice hepatic, extrahepatic antioxidant and drug metabolizing enzymes related structures

ABSTRACT In this work, the potential chemopreventive activities of Elaeagnus umbellata fruit aqueous (EUFA) and leaf aqueous (EULA) extracts focusing on the modulatory influence of xenobiotic metabolizing enzymes (XMEs), antioxidant enzymes, glucose-6-phosphate dehydrogenase (G6PD), 6-phosphogluconate dehydrogenase (6PGD), lactate dehydrogenase (LDH) activity, lipid peroxidation (LP), sulfhydryl groups were investigated in the hepatic and extrahepatic organs of Swiss albino mice (50 and 100 mg/kg body wt given orally for 14 days) and compared with BHA (0.75 % in diet). The modulatory and chemopreventive properties of two different doses EUFA and EULA were observed for cytochrome P450, cytochrome b5, sulfhydryl groups, NADPH-cytochrome P450 reductase, NADH-cytochrome b5 reductase, 7-ethoxyresorufin-deethylase and N,N-dimethylaniline N-oxidase activities in the liver and compared with BHA as a standard. The activities of glutathione S-transferase (GST) and DT-diaphorase (DTD) showed a significant increase in the kidney, forestomach, heart and brain at both doses of EUFA and EULA. The results of EULA-treated groups were found a notable increase in LDH, G6PD, 6PGD, GST and DTD activities. Superoxide dismutase level in liver, kidney and heart exhibited a significant increase at both doses of EULA. Glutathione reductase activity was a remarkable level at high dose of EUFA in liver, kidney and EULA in kidney. Both doses of EUFA were effective in inducing glutathione peroxidase activitiy in heart. The levels of LP at low and high doses of EULA-treated and EUFA-treated were effective in liver and kidney, respectively. The present results demonstrate that significant effects in the level of XMEs and antioxidant enzymes of EUFA and EULA are remarkable for modulating roles and natural chemoprevention properties and therefore is considered for a valuable natural source.

Phenolic Content and Biomolecule Oxidation Protective Activity of Globularia alypum Extracts

ABSTRACT The protective activity of methanolic (Met E) and aqueous (Aq E) extracts of Globularia alypum L. (G. alypum) against DNA, lipid and protein oxidative damage was investigated. Moreover, the scavenging, chelating, and reducing power activities of the extracts were also evaluated. Phytochemical analysis was performed to determine phenolic compounds. Results showed that Met E and Aq E were rich in phenolic compounds, and were able to scavenge DPPH˙ with IC50 values of 48.61 µg/mL and 51.97 µg/mL, respectively. In addition, both extracts were able to chelate ferrous ions. At 300 μg/mL, the chelating activity was 97.53% and 91.02%, respectively. The reducing power of these extracts was also remarkable and concentration dependent. At 100 µg/mL, both extracts inhibited lipid peroxidatin by only 42.45% and 4.03%. However, the DNA oxidation damage was inhibited dose-dependently in the presence of G. alypum extracts. At 1 mg/mL, both extracts suppressed DNA cleavage by 83%-84%. The protein oxidation was also inhibited by G. alypum extracts. At 1 mg/mL, Aq E and Met E protected BSA fragmentation by 77%-99%. The overall results suggest that G. alypum extracts exerted antioxidant activity and protect biomolecules against oxidative damage; hence it may serve as a potential source of natural antioxidants.

Phytochemical screening, anticancer and antioxidant activities of Origanum vulgare L. ssp. viride (Boiss.) Hayek, a plant of traditional usage.

BACKGROUND: A detailed phytochemical analysis of Origanum vulgare L. ssp. viride (Boiss.) Hayek was carried out and the antioxidant activities of five different crude extracts were determined. The antiproliferative activities of the extracts were determined using the xCELLigence system (Real Time Cell Analyzer). RESULTS: Differences between the essential oil and volatile organic compound profiles of the plant were shown. The main component of the essential oil was caryophyllene oxide, while the main volatile organic compounds were sabinene and eucalyptol as determined by HS-GC/MS. Phenolic contents of the extracts were determined qualitatively and quantitatively by HPLC/TOF-MS. Ten phenolic compounds were found in the extracts from O. vulgare and Origanum acutidens: rosmarinic acid (in highest abundance), chicoric acid, caffeic acid, p-coumaric acid, gallic acid, quercetin, apigenin-7-glucoside, kaempferol, naringenin and 4-hydroxybenzaldehyde. CONCLUSION: This study provides first results on the antiproliferative and antioxidant properties and detailed phytochemical screening of O. vulgare ssp. viride (Boiss.) Hayek.

Antioxidant properties of Urtica pilulifera root, seed, flower, and leaf extract.

This study was conducted to evaluate the antioxidative properties of hydroalcoholic (80%) extracts from different parts of Urtica pilulifera L. (Family Urticaceae), including leaf (UPL), flower (UPF), seed (UPS), and root (UPR). Antioxidative activity of the extracts was measured using the ferric thiocyanate method, thiobarbituric acid method, reductive potential, metal chelating, free radical, superoxide anion radical, and hydrogen peroxide scavenging activity. In addition, the results were compared with antioxidants such as tert-butylated hydroxyanisole (BHA), tert-butylated hydroxytoluene (BHT), and α-tocopherol. Total antioxidant activities of UPS, UPF, UPL, UPR, BHA, BHT, and α-tocopherol were 88.79%, 85.13%, 86.72%, 78.46%, 81.31%, 76.12%, and 46.28%, respectively. Like the antioxidant activity, the reducing power and the superoxide anion radical and free radical scavenging activities of UPL, UPF, UPS, and UPR are concentration dependent. A correlation between higher antioxidant activity and the amount of total phenolics was found in the extracts.