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CRF, Non-Selective

Cell Lines and Reagents In vitro assays were carried out using normal VERO (ECACC, No

Cell Lines and Reagents In vitro assays were carried out using normal VERO (ECACC, No. extracts. The methanol extract showed potent enzyme inhibitory activity (4.87 mg galantamine equivalent/g, 3.52 mg galantamine equivalent/g, 126.80 mg kojic acid equivalent/g, and 24.68 mg acarbose equivalent/g, for acetylcholinesterase, butyrylcholinesterase, tyrosinase, and -glucosidase, respectively) and antioxidant potential (96.52, 109.10, 154.02, and 104.85 mg trolox equivalent/g, for DPPH, ABTS, CUPRAC, and FRAP assays, respectively). Interestingly, caffeic acid-extracts showed no cytotoxicity towards VERO cell line and a weak cytotoxic potential against FaDu and SCC-25 cell lines. Interesting scientific evidence gathered from the present study support further investigation on in the view of designing and developing a novel therapeutic agent for the management of Alzheimers disease, type II diabetes, skin hyperpigmentation problems, as well as cancer. (DC.) Boiss. is used against the common cold [8]. However, to date, few information exists regarding the use of as food ingredient; in fact, to the best of our knowledge, the edible part of this plant (also known as paper pumpkinseed) is the young leaf. In particular, the raw leaves are eaten in the eastern Mediterranean as part of salads. No additional information is provided in the scientific literature regarding other uses as a food ingredient. Overall, a decoction prepared from the stem and fruits of (L.) Medik. is used against FAXF kidney stones [9], whilst powdered fruits of Boiss. are used against cattle infertility [10]. extracts were previously reported exhibiting anti-leishmanial activities on the intracellular amastigote form of the parasite and induced nitrous oxide production by human macrophages [11]. Therefore, according to the literature, the comprehensive chemical characterization, together with the description of other biological properties (such as enzyme inhibitory and/or anti-cancer potential) of the Fibigia species, is still scarce. Considering the importance of plant bioactive compounds as related to health-promoting attributes, several recent works analyzed the novel source of phytochemicals by using high-resolution targeted/untargeted mass spectrometry approaches [4,6,7]. In fact, according to the literature [12], using liquid chromatography coupled with mass spectrometry (LC-MS) is recommended to profile and then quantify antioxidant compounds (such as polyphenols) in both plant and food matrices. Therefore, the main goal of this work was to assess the potential enzyme inhibitory activity, in vitro antioxidant properties, and cytotoxicity of the ethyl acetate, methanol, and aqueous extract of was collected IDO-IN-12 in the area of Han?n village (Kastamonu, Turkey) in the summer of 2019. Taxonomic identification was performed by the botanist Dr. Ismail Senkardes (Marmara University, Department of Pharmaceutical Botany, Istanbul, Turkey), and 1 voucher specimen was deposited at the herbarium of Selcuk University (MARE-19856). The grinding of naturally dried aerial parts of the plant was carried out by a laboratory mill. For the extraction step, the maceration technique based on two different organic solvents, namely ethyl acetate (EA) and methanol. For this purpose, samples of the plant material (5 g) IDO-IN-12 were macerated with 100 mL of each solvent for 24 h at room temperature (about 25 C). Then, the solvents were evaporated under vacuum using a rotary evaporator. The aqueous extract was prepared by traditional infusion technique, and plant material (5 g) was kept with the boiled water (100 mL) for 20 min. Then the water extract was filtered and then lyophilized. All extracts were stored at +4 C until analysis. 2.2. Profiling of Bioactive Compounds in the Different Extracts To determine total phenolic and flavonoid contents of extracts, colorimetric methods were used based on our previous work [13]. In this regard, the results were finally expressed as namely gallic acid equivalents (GAE) for total phenolics and rutin equivalents (RE) for total flavonoids. Thereafter, the phytochemical analysis of each plant extract was carried out using Agilent 1200 Infinity HPLC and Agilent 6530B QTOF spectrometer (Agilent Technologies, Santa Clara, CA, USA). The conditions of the analyses were described previously [14]. The identification was based on the obtained fragmentation patterns, which were compared to the data available in the scientific literature and the Metlin database (https://metlin.scripps.edu). 2.3. Determination of Antioxidant and Enzyme Inhibitory Effects To detect antioxidant properties, several chemical assays were used, including different mechanisms, namely, radical scavenging, reducing power, and metal chelating. Trolox (TE) and ethylenediaminetetraacetic acid (EDTA) were used as standard antioxidant compounds. Obtained results were expressed as equivalents of these compounds, Grochowski, et al. [15]. To detect inhibitory effects on enzymes, colorimetric enzyme inhibition assays were used, and these assays included tyrosinase, -glucosidase, -amylase, and cholinesterases. Some standard inhibitors (galantamine, kojic acid, and acarbose) were used as positive controls. 2.4. Cell Assays 2.4.1. Cell Lines and Reagents In vitro assays were carried out using normal VERO (ECACC, No. 84113001) and cancer IDO-IN-12 FaDu (ATCC, HTB-43) and SCC-25 (ATCC, CRL-1628) cell lines. Cell media used in experiments, antibiotic supplement (Penicillin-Streptomycin Solution), and PBS (phosphate buffer saline) were.