scholarly journals Rapid Identification and Isolation of Inhibitors of Rat Lens Aldose Reductase and Antioxidant inMaackia amurensis

2017 ◽  
Vol 2017 ◽  
pp. 1-10 ◽  
Author(s):  
Set Byeol Kim ◽  
Seung Hwan Hwang ◽  
Zhiqiang Wang ◽  
Jae Myung Yu ◽  
Soon Sung Lim

Oxidative stress and aldose reductase activity have been implicated in the development of diabetic complications. In this study, the antioxidant and aldose reductase (AR) inhibitory effects ofMaackia amurensis(MA) were investigated. The ethyl acetate fraction of the MA extract showed the highest inhibitory activity in antioxidant and rat lens AR (RLAR). To identify and isolate the active components in the ethyl acetate fraction of the MA extract, high-speed countercurrent chromatography and Sephadex LH-20 column chromatography were performed and guided by an offline HPLC-ABTS assay and HPLC microfractionation AR assay. Four antioxidants, namely, piceatannol (IC50= 6.73 μM), resveratrol (IC50= 11.05 μM),trans-ferulic acid (IC50= 13.51 μM), and chlorogenic acid (IC50= 27.23 μM), and six AR inhibitors, namely, chlorogenic acid (IC50= 4.2 μM), tectoridin (IC50= 50.4 μM), genistein (IC50= 57.1 μM), formononetin (IC50= 69.2 μM), resveratrol (IC50= 117.6 μM), and daidzein (IC50= 151.9 μM), were isolated and identified. The screening results of the offline HPLC-ABTS assay and HPLC microfractionation AR assay matched the activity of isolated compounds. Thus, MA is potentially valuable for antioxidant and AR inhibitor discovery and efficient drug design for the prevention and treatment of diabetic complications.

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
Ji Hun Paek ◽  
Kuk Hyun Shin ◽  
Young-Hee Kang ◽  
Jae-Yong Lee ◽  
Soon Sung Lim

The ethyl acetate (EtOAc) soluble fraction of methanol extracts ofPerilla frutescens(P. frutescens) inhibits aldose reductase (AR), the key enzyme in the polyol pathway. Our investigation of inhibitory compounds from the EtOAc soluble fraction ofP. frutescenswas followed by identification of the inhibitory compounds by a combination of HPLC microfractionation and a 96-well enzyme assay. This allowed the biological activities to be efficiently matched with selected HPLC peaks. Structural analyses of the active compounds were performed by LC-MSn. The main AR inhibiting compounds were tentatively identified as chlorogenic acid and rosmarinic acid by LC-MSn. A two-step high speed counter current chromatography (HSCCC) isolation method was developed with a solvent system of n-hexane-ethyl acetate-methanol-water at 1.5 : 5 : 1 : 5, v/v and 3 : 7 : 5 : 5, v/v. The chemical structures of the isolated compounds were determined by1H- and13C-nuclear magnetic resonance spectrometry (NMR). The main compounds inhibiting AR in the EtOAc fraction of methanol extracts ofP. frutescenswere identified as chlorogenic acid (2) (IC50= 3.16 μM), rosmarinic acid (4) (IC50= 2.77 μM), luteolin (5) (IC50= 6.34 μM), and methyl rosmarinic acid (6) (IC50= 4.03 μM).


Molecules ◽  
2021 ◽  
Vol 26 (9) ◽  
pp. 2555
Author(s):  
Sang Koo Park ◽  
Yoon Kyung Lee

Using natural products as antioxidant agents has been beneficial to replace synthetic products. Efforts have been made to profile the antioxidant capacities of natural resources, such as medicinal plants. The polyphenol content of Himalayan rhubarb, Rheum emodi wall, was measured and the antioxidant activity was determined using DPPH and ABTS+ assay, and the oxidative stress was assessed using SOD enzymatic assay. Five different solvent fractions, n-hexane, n-butanol, ethyl acetate, dichloromethane, and water, were used for screening the antioxidant capacity in effort to determine the optimum extraction solvent. The total phenolic contents for R. emodi fractions ranged from 27.76 to 209.21 mg of gallic acid equivalents (GAE)/g of dry weight. DPPH and ABTS+ assay results are presented into IC50 values, ranged from 21.52 to 2448.79 μg/mL and 90.25 to 1718.05 μg/mL, respectively. The ethyl acetate fraction had the highest antioxidant activity among other fractions. Also, n-butanol and water fractions showed significantly lower IC50 values than the positive control in DPPH radical scavenging activity. The IC50 values of SOD assay of fractions ranged from 2.31 to 64.78 μg/mL. A similar result was observed with ethyl acetate fraction showing the highest SOD radical scavenging activity. The study suggests that the ethyl acetate fraction of R. emodi possess the strongest antioxidant activity, thus the most efficient in extracting antioxidant contents. Moreover, a highly significant correlation was shown between total polyphenol content and antioxidant activity screening assays. The compounds related to the antioxidant activity of R. emodi were identified to myricitrin, myricetin 3-galloyl rhamnoside, and myricetin, which have not been reported in studies about R. emodi before.


2014 ◽  
Vol 86 (2) ◽  
pp. 881-888 ◽  
Author(s):  
MARIANA PIANA ◽  
ALINE A. BOLIGON ◽  
THIELE F. DE BRUM ◽  
MARINA ZADRA ◽  
BIANCA V. BELKE ◽  
...  

The antioxidant capacity of the crude extract and fractions ofTabernaemontana catharinensis fruits and branches, was evaluated by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method and the content of polyphenols, flavonoids, alkaloids and condensed tannins were determined by the spectrophotometric method. The ethyl acetate fraction of the fruits and the n-butanol fraction of the branches showed IC50 of 181.82 µg/mL and 78.19 µg/mL, respectively. All fractions were analyzed by high performance liquid chromatography (HPLC), in the branches were quantified chlorogenic acid in the chloroform (8.96 mg/g), ethyl acetate (4.31 mg/g) and n-butanol (3.33 mg/g) fractions; caffeic acid in the ethyl acetate (5.24 mg/g) and n-butanol (1.81 mg/g); gallic acid (0.52 mg/g) in the n-butanol. In the fruits, chlorogenic acid in the chloroform (1.67 mg/g); rutin in the ethyl acetate (3.45 mg/g) and n-butanol (8.98 mg/g) fractions. The present study showed that these quantified compounds can contribute to antioxidant capacity which was higher in the branches than in the fruits.


Author(s):  
Le Quy Thuong ◽  
Bach Tuyet Mai ◽  
Nguyen Minh Chau ◽  
Le Thi Phuong Hoa ◽  
Nguyen Quang Huy

Typhonium flagelliforme is a medicinal plant that has variety of uses. In medicinal traditional T. flagelliforme is used to treatment cough, headache, stomach pain chronic, and tracheitis. Moreover, use fresh bulbs treatment furuncle, the bites of poisonous insects. The active components in T. flagelliforme are flavonoids. In this study, the T. flagelliforme extract was obtained by methanol  to determine the chemical composition. Then, The extracts of methanol are extracted with polarization increases gradually solvents such as haxane, dichloromethane and ethyl acetate. Determination of antioxidant activity, cytotoxic activity of extracted fractions. Results obtained showed that the chemical compositions by the qualitative reaction preliminary were identified from T. flagelliforme containing reducing sugars, amino acids, organic acids, flavonoids, alcaloids, sterols. The antioxidant capacity of the ethyl acetate fraction reached 94.76 μg/ml, 10 times higher than the positive control is Quercetin. Cytotoxic activity of the haxane and diclomethane extracted fractions from T. flagelliforme exhibited cytotoxic activity on all three experimental cancers cell lines: KB, HepG2, Lu after 72h of culture with IC50 values ​​range from 92.8 to 107.76 μg/ml. From dichlomethane extracted of T. flagelliforme was purified TF1 as Stigmast-4-en- 3-on.    


2021 ◽  
Vol 33 (5) ◽  
pp. 977-982
Author(s):  
Deepak Singh Janoti ◽  
Kumud Upadhyaya

The present study is based on the selection of extract and fraction of Swertia chirata plant for the antioxidant potential with HPLC fingerprinting, which includes the simultaneous detection and quantification of four analytical markers protocatechuic acid (PCA), swertiamarin (SM), mangiferin (MF) and amarogentin (AG) by HPLC-DAD. The yield of water extract (SWA), hydroalcoholic extract (SHA) and fractions of hydroalcoholic extracts were evaluated for their antioxidant potential against 2,2-diphenyl-1-picrylhydrazyl-hydrate free radical assay (DPPH assay), 2,2′-azino-bis(3- ethylbenzothiazoline-6-sulfonic acid radical scavenging assay (ABTS assay), total reducing assay (TRA), ferric reducing antioxidant potential assay (FRAP assay), total antioxidant capacity assay (TAC assay). The hydroalcoholic extracts (SHA) can be a better choice as compared to water extract (SWA) due to higher yield of extract (13.680 ± 0.548%) and higher antioxidant activity against DPPH assay, ABTS assay, TRA assay, FRAP assay and TAC assay. In hydroalcoholic extract (SHA), ethyl acetate fraction (SEA) showed most potent activity against DPPH (IC50 = 0.008 ± 0.002 mg/mL) and ABTS (0.025 ± 0.001 mg/mL). n-Hexane fraction of SHA showed higher FRAP (28.664 ± 3.153 μmol/mL) and TAC (3.263 ± 0.325 μmol/mL) value (equivalent to ascorbic acid in μmol/mL) but showed very low yield (0.468 ± 0.018%), SBU showed higher TRA value (0.413 ± 0.309 mg/mL). The ethyl acetate fraction (SEA) can be a choice for antioxidant as it showed second highest FRAP (19.547 ± 2.119 μmol/mL) and TAC (2.750 ± 0.466 μmol/mL) with better yield (2.473 ± 0.594%) as compared to n-hexane (SH) fractions (0.468 ± 0.018%).


Molecules ◽  
2019 ◽  
Vol 24 (15) ◽  
pp. 2709 ◽  
Author(s):  
Zhiqiang Wang ◽  
Shigang Shen ◽  
Ze Cui ◽  
Hailiang Nie ◽  
Dandan Han ◽  
...  

Aldose reductase (AR) is a drug target for therapies to treat complications caused by diabetes mellitus, and the development of effective AR inhibitors (ARIs) of natural origin is considered to be an attractive option for reducing these complications. In this research, the rat lens AR (RLAR) inhibitory activity of evening primrose (Oenothera biennis) seeds was investigated for the first time. In our results, the 50% (v/v) methanol extract of evening primrose seeds exhibits excellent RLAR inhibitory activity (IC50 value of 7.53 μg/mL). Moreover, after enrichment of its bioactive components, the ARIs are more likely to be present in the ethyl acetate fraction of 50% (v/v) methanol extract (EME) of evening primrose seeds, which exhibits superior RLAR inhibitory activity (IC50 value of 3.08 µg/mL). Finally, gallic acid (1), procyanidin B3 (2), catechin (3), and methyl gallate (4) were identified as the major ARIs from the EME by affinity-based ultrafiltration-high-performance liquid chromatography and were isolated by high speed countercurrent chromatography, with gallic acid (11.46 µmol/L) and catechin (14.78 µmol/L) being the more potent inhibitors of the four ARIs identified. The results demonstrated that evening primrose seeds may be a potent ingredient of ARIs.


2020 ◽  
Vol 2020 ◽  
pp. 1-8
Author(s):  
Zahra Abbasi ◽  
Gholamali Jelodar ◽  
Bita Geramizadeh

Background. Increased activity of aldose reductase (AR) is one of the mechanisms involved in the development of diabetic complications. Inhibiting AR can be a target to prevent diabetes complications. This study is aimed at evaluating the effect of cyclohexane (CH) and ethanol extracts (ET) of walnut leaves on AR activity in the lens and testis of diabetic rats. Methods. Fifty-six male rats classified into seven groups as control and treatment groups and treated for 30 days. The treatment groups were treated with different concentrations of ET and CH. The diabetic control (DC) group was exposed to streptozotocin. AR activity was measured in the lens and testis. The expression of AR in the testis was evaluated by the immunohistochemistry method. Results. Both extracts significantly reduced the AR activity (ng/mg of tissue protein) in the testis (0.034±0.004, 0.038±0.010, and 0.040±0.007 in the treatment groups vs. 0.075±0.007 in the DC group) and lens (1.66±0.09, 2.70±0.47, and 1.77±0.20 in the treatment groups vs. 6.29±0.48 in the DC group) of the treatment group compared to those of the DC group (P<0.05). AR expression in the testes of the treatment groups was decreased compared with that of the DC group (P<0.0001). Conclusion. Walnut leaf extracts can reduce the activity and localization of AR in the testes and its activity in the lens of diabetic rats.


2011 ◽  
Vol 34 (5) ◽  
pp. 847-852 ◽  
Author(s):  
Chan-Sik Kim ◽  
Junghyun Kim ◽  
Yun Mi Lee ◽  
Eunjin Sohn ◽  
Kyuhyung Jo ◽  
...  

2018 ◽  
Vol 2018 ◽  
pp. 1-6 ◽  
Author(s):  
Lei Fang ◽  
Hua Zhang ◽  
Jie Zhou ◽  
Yanling Geng ◽  
Xiao Wang

An efficient method using high-speed counter-current chromatography (HSCCC) coupled with DPPH-HPLC assay has been developed for rapid screening and preparative isolation of antioxidants from ethyl acetate fraction ofAlpinia officinarumHance. Target-guided by DPPH-HPLC assay, two antioxidants, galangin and kaempferide, were targeted and further separated with purities of 99.3% and 98.5% by HSCCC using petroleum ether–ethyl acetate–methanol–water (0.8 : 1 : 1 : 0.8,v/v) as the solvent system. The antioxidant activities of galangin and kaempferide were further evaluated by measuring their inhibiting effects on superoxide anion radical, hydroxyl radical, and hydrogen peroxide in different luminol chemiluminescence (CL) systems. As a result, galangin and kaempferide both showed potent antioxidant activities. Results of the present study indicated that the combinative method by offline coupling DPPH-HPLC and HSCCC could be widely applied for rapid screening and isolation of antioxidants from complex TCM extract.


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