Turk J Pharm Sci 2018;15(1):97-102 DOI: 10.4274/tjps.82473 ORIGINAL ARTICLE Screening Effects of Methanol Extracts of Diplotaxis tenuifolia and Reseda lutea on Enzymatic Antioxidant Defense Systems and Aldose Reductase Activity Diplotaxis tenuifolia ve Reseda lutea Metanol Özütünün Antioksidan Savunma Sistemi Enzimleri ve Aldoz Redüktaz Aktivitesi Üzerinde Olan Etkisinin İncelenmesi Khalid Sharro ABDALRAHMAN1, Merve Gülşah GÜNEŞ1, Naznoosh SHOMALI1*, Belgin Sultan İŞGÖR2, Özlem YILDIRIM1 1Ankara University, Faculty of Science, Department of Biology, Ankara, Turkey 2Atılım University, Faculty of Engineering, Department of Chemical Engineering and Applied Chemistry, Ankara, Turkey ABSTRACT Objectives: The aim of the study was to investigate the effects of methanol extracts from the flowers and leaves of Diplotaxis tenuifolia and Reseda lutea on the activity of AR, CAT, GST, and GPx. Materials and Methods: Total phenolic and flavonoid contents of the plant samples were evaluated using Folin-Ciocalteu reagent and aluminum chloride colorimetric methods. Also, the effects of extracts on CAT, GST, GPx, and AR enzyme activities were investigated using kinetic assays. Results: The highest phenolic and flavonoid contents were detected in the methanol extract of D. tenuifolia leaves with 144.49±0.29 mg gallic acid equivalent/L and 250.485±0.002 quercetin equivalent/L, respectively. The best activity profile for GST and GPx were observed in the extract of leaves belonging to D. tenuifolia with IC50 values of 121±0.05 and 140±0.001 ng/mL, respectively. According to the results, methanol extracts from leaves of R. lutea and D. tenuifolia showed no significant activity potential on AR. Moreover, none of the studied extracts demonstrated any reasonable CAT activation potential. Conclusion: The results indicated that leaves of D. tenuifolia had good effect on the antioxidant enzymatic defense system, which it makes it a good constituent of the daily diet. Key words: Diplotaxis tenuifolia, Reseda lutea, antioxidant enzymes, aldose reductase ÖZ Amaç: Bu çalışmada Diplotaxis tenuifolia ve Reseda lutea’nın çiçek ve yapraklarından elde edilmiş olan metanol özütlerinin AR, CAT, GST ve GPx enzimlerinin aktiviteleri üzerinde olan etkilerinin araştırılması amaçlanmıştır. Gereç ve Yöntemler: Bu çalışmada, bitki örneklerinin toplam fenolik ve flavonoid içeriği; Folin-Ciocalteu ve alüminyum klorür reaktiflerinin yardımıyla kolorimetrik yöntemlerle değerlendirilmiştir. Ayrıca özütlerin CAT, GST, GPx ve AR enzimlerinin aktiviteleri üzerindeki etkileri kinetik analizler ile araştırılmıştır. Bulgular: En yüksek miktarda fenolik ve flavanoid içeriği sırasıyla 144.49±0.29 mg gallik asit eş değeri/L ve 250.485±0.002 quercetin eş değeri/L tespit edilmiştir. GST ve GPx için en iyi aktivite profilleri sırasıyla 121±0.05 ve 140±0.001 ng/mL IC50 değerleri ile D. tenuifolia yaprak özütünde gözlemlenmiştir. Elde edilen sonuçlara göre, R. lutea ve D. tenuifolia’nın yapraklarından elde edilen metanol özütleri, AR enzimi üzerinde önemli ölçüde bir aktivite potansiyeli göstermemiştir. Bununla beraber, çalışılmış olan çiçek ve yaprak özütlerinin hiçbiri yeterli düzeyde CAT aktivasyonu gösterememiştir. Sonuç: Çalışma sonucunda, D. tenuifolia’nın yapraklarının antioksidan enzimatik savunma sistemi üzerinde iyi bir etkiye sahip olduğu gösterilmiştir. Bu sebeple günlük diyet için iyi bir besin kaynağı olarak kabul edilebilir. Anahtar kelimeler: Diplotaxis tenuifolia, Reseda lutea, antioksidan enzimler, aldoz redüktaz *Correspondence: E-mail: [email protected], Phone: +90 554 281 63 36 ORCID-ID: orcid.org/0000-0002-7366-1569 Received: 18.01.2017, Accepted: 16.03.2017 ©Turk J Pharm Sci, Published by Galenos Publishing House. 97 98 ABDALRAHMAN et al. Screening Effects of Methanol Extracts of Diplotaxis tenuifolia INTRODUCTION Phytochemical studies show that the aerial parts of D. tenuifolia Reactive oxygen species (ROS) is a term used to describe a contain significantly high concentration of flavonoids, tannins, number of reactive molecules and free radicals derived from glucosinolates, sterols, and vitamin C.9 molecular oxygen, which are generated by all aerobic species. The genus Reseda is one of the herbs in the Resedaceae family. These molecules are generated as by-products during the In Turkey, this genus is represented by 15 species including mitochondrial electron transport of aerobic respiration or Reseda lutea L. and Reseda luteola L. It is known as yellow by oxidoreductase enzymes and metal catalyzed oxidation. mignonette or wild mignonette and has economic importance. In normal physiologic conditions, a number of defense It is widely used in the carpet and rug industry as a source mechanisms have evolved to provide a balance between the of natural dye due to its high luteolin content. In addition to production and removal of ROS, but alterations of the balance its staining properties, luteolin has attracted great scientific between ROS production and the capacity to detoxify reactive interest because of its pharmacologic activities. Luteolin intermediates lead to oxidative stress. It has been caused to a displays numerous anti-inflammatory effects at micromolar wide variety of states, processess and metabolic diseases such concentrations, which cannot be completely explained by its as heart disease, severe neural disorders such as Alzheimer’s antioxidant capacities. In addition, phytochemical analysis of and Parkinson’s, and some cancers.1,2 Under oxidative stress, aerial parts of R. lutea has shown the presence of flavonoid, an organism has a variety of defense mechanisms to prevent anthocyanin, and glucosides.10 or neutralize negative ROS effects. These are mainly based on The aim of the present study was to evaluate the total amount enzymes such as catalase (CAT), superoxide dismutase (SOD), of the phenolic and flavonoid contents of methanol extract glutathione peroxidase (GPx), glutathione reductase (GR), and obtained from the flowers and leaves of D. tenuifolia and R. lutea glutathione-S-transferase (GST) or non-enzymatic components and to determine their effects on the activity of AR, CAT, GST, 3 such as vitamin E, vitamin C, glutathione, and flavonoids. and GPx. These enzymes play critical roles in the antioxidant GST is one of the phase II enzymes and plays a critical role defense system. in the detoxification and metabolism of many xenobiotic 4 compounds. GPx has an important role as a catalyst in the EXPERIMENTAL reduction of hydro peroxides, including hydrogen peroxides (H2O2), by using GSH. GPx also functions to protect the cell Chemicals materials from oxidative damage. Several studies related dysfunctional In this study, 4-aminoantipyrine, H2O2 and sodium azide (NaN3) GPx with cancer.5 CAT is a very important enzyme of living were provided by Acros, USA. Ethylenediaminetetraacetic acid organisms, which catalyzes the decomposition of H2O2 to water (EDTA), Folin-Ciocalteu reagent, reduced glutathione (GSH), and oxygen. Aldose reductase (AR) is a nicotinamide adenine GR, horseradish peroxidase, CAT, gallic acid (GA), and quercetin dinucleotide phosphate (NADPH)-dependent enzyme and it hydrate were supplied by Sigma Aldrich, Germany. Lithium has been implicated in the formation of cancer and diabetic sulphate (Li2SO4) and NADPH were purchased from Gerbu, complications such as retinopathy, neuropathy, nephropathy, Germany. All other chemicals used were analytical grade and and cardiovascular disorders.6 provided by Sigma Aldrich, Germany. Plants synthesize a vast range of organic compounds that are Plant materials traditionally classified as primary and secondary metabolites. Plant samples of D. tenuifolia and R. lutea were harvested in Primary metabolites are compounds that have essential roles July 2010 from Ankara, Turkey, and were authenticated by associated with photosynthesis, respiration, growth, and Prof. Dr. Fatmagül Geven, in the Department of Biology, development. Other phytochemicals that accumulate in high Ankara University. The plant specimens with their localities concentrations in some species are known as secondary and the necessary field records were recorded and numerated metabolites, which possess antioxidant activity. Antioxidant as voucher specimen numbers. The voucher numbers of compounds found in different parts of plants involve phenolics, D. tenuifolia and R. lutea were FG-2010-10 and FG-2010-13, flavonoids, alkaloids, glycosides, tocopherols, carotenoids, and respectively. They were deposited in the herbarium department ascorbic acid. These are structurally diverse and many are at Ankara University. distributed among a very limited number of species within the plant kingdom.7 Secondary metabolite compounds have played Extraction of plant an important role in treating and preventing human diseases. Different parts of fresh plant samples (flowers and leaves) were They are important sources for new drugs and are also washed with tap water and dried at room temperature before suitable lead compounds for further modification during drug analysis. For methanol extraction, 2 g of dried samples were 4 development. weighed and ground into a fine powder with liquid nitrogen, then Diplotaxis tenuifolia (L.) DC., commonly known as ‘wild rocket’, mixed with 20 mL methyl alcohol at room temperature in 160 rpm belongs to the Brassicaceae family. It was originally found as for 24 h. The obtained extract was filtered over Whatman No. 1 a crop in Mediterranean and Middle Eastern countries and paper and the filtrate was collected. Methanol