In vitro Antioxidant, Cytotoxic and Antidiabetic Activity of Protein Hydrolysates Prepared from Grass Turtle (Chinemys reevesii)

Research background. Cardiovascular diseases and diabetes are the biggest causes of death globally. Bioactive peptides derived from many food proteins using enzymatic proteolysis and food processing perform a positive impact on the prevention of these diseases. The bioactivity of grass turtle muscle proteins and their enzymatic hydrolysates has not received much attention, thus this study aims to investigate the antioxidant, antidiabetic, and cytotoxic activities of the enzymatic hydrolysates of grass turtle muscle protein. Experimental approach. Grass turtle muscle(s) was hydrolysed using four proteolytic enzymes (Alcalase (AH), Flavourzyme (FH), Trypsin (TH), and Bromelain (BH)). The degree of hydrolysis was measured. High performance liquid chromatography (HPLC) was conducted to explore their amino acid profiles and molecular mass distribution of the hydrolysates. The antioxidant activities were evaluated using various in vitro tests, including 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), Hydroxyl radical (•OH) radical scavenging activities, reducing capacity, chelating (Fe2+), and lipid peroxide inhibition activity. Antidiabetic activity was evaluated against α-amylase inhibition and α-Glucosidase inhibition assay. Besides, cytotoxic effect of hydrolysates on human colon cancer (HT-29) cells was assessed. Results and conclusions. The amino acid composition of the hydrolysates revealed that higher contents of glutamic, aspartic, lysine, hydroxyproline, and hydrophobic amino acids. AH showed significantly the highest inhibition of lipid peroxidation. FH showed the highest lipid peroxidation, strongest radical scavenging activity of DPPH (68.32 %), ABTS (74.12 %), FRAP (0.30 unit), α-glucosidase (61.80 %) and cytotoxic effect on cancer cells (82.26 %) at 550 µg/mL for HT-29 cell line. TH and BH were showed significant (p<0.05) hydroxyl radical activity (92.70 %) and Fe2+ metal chelating (63.29 %), respectively. The highest α-amylase (76.89 %) inhibition was recorded by BH and FH. Novelty and scientific contribution. Enzymatic grass turtle muscle(s) proteins hydrolysates achieved great antioxidant, cytotoxic, and antidiabetic activity. The findings of this study indicated that the bioactive hydrolysates /peptides from grass turtle muscle(s) protein could be potential candidate as ingredients in pharmaceuticals and functional food formulations.

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In vitro Antioxidant Study and Determination of Flavonoids, Flavonols, Total Phenolic and Proanthocyanidins Content of Grewia abutilifolia Leaf Extracts

Phytothérapie ◽

10.3166/phyto-2018-0102 ◽

2019 ◽

Vol 18 (3-4) ◽

pp. 140-147

Author(s):

R. Salam ◽

R. Rafe

Keyword(s):

Lipid Peroxidation ◽

Hydroxyl Radical ◽

Total Phenolic Content ◽

Phenolic Content ◽

Radical Scavenging ◽

Total Phenolic ◽

Inhibition Activity ◽

Aqueous Fraction ◽

Lipid Peroxidation Inhibition

Grewia abutilifolia (Tiliaceae) tree is known for its uses in traditional medicine due to its various degrees of ethno-pharmacological activities. In this study, Grewia abutilifolia was evaluated for its in vitro antioxidant activities; further, flavonoids, flavonols, total phenolic content and proanthocyanidins content were determined to justify its uses against various oxidative stresses.We evaluated antioxidant activity by determining total antioxidant, Fe3+ reducing power, DPPH (2,2-diphenyl-1-picrylhydrazyl) and hydroxyl radical scavenging, and lipid peroxidation inhibition activity. Aqueous fraction (AQF) contained highest total phenolic content (238.56 mg of gallic acid equivalent/g). Crude methanolic extract (CHE) contained maximum flavonoid (211.54 mg) and flavonols (380.25 mg) content equivalent to quercetin which showed significant reducing activity. Proanthocyanidins content was highest in petroleum ether fraction (PEF), 45.89 mg of catechin equivalent per gram. PEF showed maximum activity in DPPH scavenging assay (IC50 = 3.82 ± 0.055 μg/ml) and in hydroxyl radical scavenging assay (IC50, concentration required to inhibit the process by half, of 6.45 ± 1.297 μg/ml). Lipid peroxidation inhibition activity was also performed in the test chloroform fraction (CLF), which showed the highest inhibition (IC50 = 15.62 ± 1.31 μg/ml). These results suggested its potentiality against various diseases related to oxidative stress.

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Cytotoxic effect of glycoprotein isolated from Solanum nigrum L. through the inhibition of hydroxyl radical-induced DNA-binding activities of NF-kappa B in HT-29 cells

Environmental Toxicology and Pharmacology ◽

10.1016/j.etap.2004.02.003 ◽

2004 ◽

Vol 17 (1) ◽

pp. 45-54 ◽

Cited By ~ 13

Author(s):

Kyung-Sun Heo ◽

Sei-Jung Lee ◽

Kye-Taek Lim

Keyword(s):

Dna Binding ◽

Hydroxyl Radical ◽

Cytotoxic Effect ◽

Solanum Nigrum ◽

Nf Kappa B ◽

Solanum Nigrum L ◽

Ht 29

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Nanostructured Ethosomal Gel Loaded with Arctostaphylos uva-ursi extract; In-vitro/In-vivo Evaluation as a Cosmeceutical Product

Current Drug Delivery ◽

10.2174/1567201818666210729111026 ◽

2021 ◽

Vol 18 ◽

Author(s):

Nayla Javed ◽

Shakeel Ijaz ◽

Naveed Akhtar ◽

Haji Muhammad Shoaib Khan

Keyword(s):

Zeta Potential ◽

Biological Activities ◽

Skin Permeation ◽

Radical Scavenging ◽

Reducing Power ◽

Potential Candidate ◽

Skin Rejuvenation ◽

In Vivo Evaluation

Background: Arctostaphylos uva-ursi (AUU) being rich in polyphenols and arbutin is known to have promising biological activities and can be a potential candidate as a cosmaceutical. Ethosomes encourage the formation of lamellar-shaped vesicles with improved solubility and entrapment of many drugs including plant extracts. Objective: The objective of this work was to develop an optimized nanostructured ethosomal gel formulation loaded with AUU extract and evaluated for skin rejuvenation and depigmentation. Methods: AUU extract was tested for phenolic and flavonoid content, radical scavenging potential, reducing power activity, and in-vitro SPF (sun protection factor) estimation. AUU loaded 12 formulations were prepared and characterized by SEM (scanning electron microscopy), vesicular size, zeta potential, and entrapment efficiency (%EE). The optimized formulation was subjected to non-invasive in-vivo investigations after incorporating it into the gel system and ensuring its stability and skin permeation. Results: Ethosomal vesicles were spherical in shape and Zeta size, zeta potential, PDI (polydispersity index), % EE and in-vitro skin permeation of optimized formulation (F3) were found to be 114.7nm, -18.9mV, 0.492, 97.51±0.023%, and 79.88±0.013% respectively. AUU loaded ethosomal gel formulation was stable physicochemically and exhibited non-Newtonian behavior rheologically. Moreover, it significantly reduced skin erythema, melanin as well as sebum level and improved skin hydration and elasticity. Conclusion: A stable AUU based ethosomal gel formulation could be a better vehicle for phytoextracts than conventional formulations for cosmeceutical applications such as for skin rejuvenation and depigmentation etc.

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IN VITRO - ANTIOXIDANT AND PRELIMINARY PHYTOCHEMICAL ANALYSIS OF AEGLE MARMELOS (L.) CORREA. LEAF EXTRACT

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2017.v10i6.17808 ◽

2017 ◽

Vol 10 (6) ◽

pp. 367

Author(s):

Samidha M Pawaskar ◽

Sasangan Kc

Keyword(s):

Hydroxyl Radical ◽

Leaf Extract ◽

Radical Scavenging ◽

Reducing Power ◽

Total Phenolic ◽

Phytochemical Analysis ◽

Aegle Marmelos ◽

Leaf Powder ◽

Phenolic And Flavonoid

Objective: In this study, the leaf powder of Aegle marmelos (L.) Correa. was subjected to preliminary phytochemical and in vitro antioxidant analysis. Methods: The freshly prepared plant leaf extract was subjected to preliminary phytochemical screening, which revealed the presence of alkaloids, tannins, saponins, flavonoids, glycosides, phenolic compounds, terpenoids, and steroids. Reducing power, superoxide (SO) anion radical, nitric oxide (NO) radical, and hydroxyl radical scavenging assays were carried out to evaluate the antioxidant potential of the methanolic leaf extract of this plant. The amounts of total phenolic and flavonoid compounds were also determined. Results: This study has revealed that the A. marmelos (L.) Correa. leaf extract showed considerably high amounts of most of the phytochemicals, total antioxidant capacity, total phenolic, and flavonoid content. The study also indicated that the A. marmelos (L.) Correa. showed comparatively good scavenging activity, i.e., inhibition of hydroxyl radical, NO and SO anion scavenging and reducing power activities when compared with the respective standards. Conclusion: The leaf powder of A. marmelos (L.) Correa. can be used as easily accessible source of natural antioxidant and as a possible food supplement or in pharmaceutical industry.

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In vitro inhibitory potentials of ethanolic extract of Moringa oleifera flower against enzymes activities linked to diabetes

Journal of Herbmed Pharmacology ◽

10.34172/jhp.2021.48 ◽

2021 ◽

Vol 10 (4) ◽

pp. 408-414

Author(s):

Oluwaseun Ruth Olasehinde ◽

Olakunle Bamikole Afolabi ◽

Benjamin Olusola Omiyale ◽

Oyindamola Adeniyi Olaoye

Keyword(s):

Free Radical ◽

Moringa Oleifera ◽

Antioxidant Properties ◽

Radical Scavenging ◽

Ethanolic Extract ◽

Antidiabetic Activity ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Antioxidant Power

Introduction: Diabetes mellitus (DM) has been recognized as the seventh leading cause of global mortality; however, researchers seek alternative means to manage the menace. The current study sought to investigate antioxidant potentials, α-amylase, and α-glucosidase inhibitory activities of ethanolic extract of Moringa oleifera flower in vitro. Methods: Antioxidant properties of the extract were appraised by assessing its inhibition against 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl (OH•), and hydrogen peroxide (H2O2) free radicals, as well as ferric reducing antioxidant power (FRAP), the antidiabetic activity was evaluated by α-amylase and α-glucosidase inhibition.Results: In this study, ethanolic extract of M. oleifera flower demonstrated a significant (P < 0.05) inhibition against DPPH free radical (43.57–83.56%) in a concentration-dependent manner, while FRAP (101.76 ± 1.63 mg/100 g), OH• scavenging ability (71.62 ± 0.95 mg/100 g), and H2O2 free radical scavenging capacity (15.33 ± 1.20 mg/100 g) were also observed. In the same manner, ethanolic extract of M. oleifera flower revealed a significant (P < 0.05) inhibition against α-amylase (IC50= 37.63 mg/mL) and α-glucosidase activities (IC50= 38.30 mg/mL) in the presence of their respective substrates in a concentration-dependent manner in comparison with acarbose. Conclusion: Ethanoic extract of M. oleifera flower could be useful as an alternative phytotherapy in the management of DM, having shown a strong antioxidative capacity and substantial inhibition against the activities of key enzymes involved in carbohydrate hydrolysis in vitro.

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EVALUATION AND COMPARISON OF CYTOTOXIC EFFECT OF VILAZODONE HYDROCHLORIDE WITH 5-FLUOROURACIL IN HT-29 BOWEL CANCER CELL LINE

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2019.v12i2.29097 ◽

2019 ◽

pp. 124-127

Author(s):

LATHA PRIYA A ◽

ANUSHA D ◽

DARLING CHELLATHAI K ◽

HEMALATHA A ◽

JEGAN MOHAN Y

Keyword(s):

Cell Line ◽

Cytotoxic Effect ◽

Depressive Disorders ◽

Cancer Cell Line ◽

Test Sample ◽

Standard Drug ◽

Drug Sample ◽

Diphenyl Tetrazolium Bromide ◽

Ht 29

Objectives: Vilazodone hydrochloride is a novel selective serotonin reuptake inhibitor (SSRI) used to treat major depressive disorders. There are only sparse data available to know about the SSRI’s and its association with colon cancer. This study aims to evaluate and compare the in vitro cytotoxic effect of vilazodone with 5-fluorouracil (5-FU) in HT-29 cell line. Methods: Cell viability was tested by the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay (Mosmann, 1983). Test sample and standard drug in variable concentrations were added to the HT-29 cell lines for incubation over 24 h under ideal conditions. After washing the test and standard drug sample from the well with saline, MTT was added and incubated for 4 h. Dimethyl sulfoxide of 1 ml was added in all wells after incubation with MTT. The absorbance at 570 nm was measured with an ultraviolet - spectrophotometer. Results: The values were tabulated, and the graph was plotted to find the IC-50 value (inhibitory concentration at 50%) which was struck at 28.5 μg/ml and12. 8 μg/ml for vilazodone hydrochloride and 5-FU, respectively. Conclusion: The results show that vilazodone hydrochloride has good anticancer property comparable with 5-FU, which would probably play a role as a cytotoxic agent in tumor cells. The proposed mechanism of action could be by activation of caspase-3 enzyme, thereby increasing apoptosis and indicates its use in coexisting depression and colon carcinoma. Other mechanism includes suppression of oncogene p53, which can be confirmed by future studies.

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IN VITRO ANTIOXIDANT AND ALPHA-AMYLASE, ALPHA-GLUCOSIDASE INHIBITORY ACTIVITY OF FLOWERS OF CALOTROPIS GIGANTEA

INDIAN DRUGS ◽

10.53879/id.51.10.10104 ◽

2014 ◽

Vol 51 (10) ◽

pp. 38-42

Author(s):

N. K Choudhary ◽

◽

J Dwivedi ◽

S Sharma

Keyword(s):

Nitric Oxide ◽

Radical Scavenging Activity ◽

Radical Scavenging ◽

Chloroform Extract ◽

Antidiabetic Activity ◽

Scavenging Activity ◽

Dpph Radical ◽

Dpph Radical Scavenging Activity ◽

Dpph Radical Scavenging

The present investigations were carried out to evaluate the in vitro antioxidant as well as antidiabetic activity of flowers of Calotropis gigantea. Different extracts (petroleum ether, chloroform and ethanolic extract) were prepared using successive solvent extraction method (soxhlet) and screened for its in vitro antioxidant activity using Diphenyl picryl hydrazyl (DPPH) radical scavenging activity, ABT S radical cation decolorization assay and nitric oxide (NO) radical scavenging activity and IC50 were calculated. In vitro antidiabetic activity was studied using α – amylase and α – glucosidase inhibitory assay. Chloroform extract, among the three extracts (i.e. petroleum ether, chloroform and ethanolic), showed maximum antioxidant activity with IC50 value of 151.23µg/ml, 73.56 µg/ml and 158.92µg/ml against DPPH radical scavenging activity, ABTS radical cation decolorization assay and nitric oxide (NO) radical scavenging activity respectively. The chloroform extract produced a significant in vitro antidiabetic activity with IC50 of 52.3µg/ml 18.2µg/ml against α – amylase and α – glucosidase enzymes but less inhibitory effect than standard acarbose.

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Evaluation of Antioxidant and Anti-Inflammatory Activity of Anthocyanin-Rich Water-Soluble Aronia Dry Extracts

Molecules ◽

10.3390/molecules25184055 ◽

2020 ◽

Vol 25 (18) ◽

pp. 4055 ◽

Cited By ~ 1

Author(s):

Mariusz Banach ◽

Magdalena Wiloch ◽

Katarzyna Zawada ◽

Wojciech Cyplik ◽

Wojciech Kujawski

Keyword(s):

Lipid Peroxidation ◽

Interleukin 1 ◽

Radical Scavenging ◽

Water Soluble ◽

Anthocyanin Content ◽

Necrosis Factor Alpha ◽

Paramagnetic Resonance ◽

Markers Of Inflammation ◽

Radical Scavenging Properties

Aronia fruits contain many valuable components that are beneficial to human health. However, fruits are characterized by significant variations in chemical composition dependent on the growing conditions and harvesting period. Therefore, there is a need to formulate the extracts with a precisely defined content of health-promoting substances. Aronia dry extracts (ADE) were prepared from frozen pomace applying water extraction, followed by purification and spray-drying. Subsequently, the content of anthocyanins, phenolic acids, and polyphenols was determined. The high-quality chokeberry pomace enabled obtaining extracts with anthocyanin content much higher than the typical market standards. Moreover, it was found that the antioxidant capacity of aronia extracts exceeded those found in other fruit preparations. Antioxidant and free-radical scavenging properties were evaluated using a 2,2′-diphenyl-1-picrylhydrazyl using Electron Paramagnetic Resonance (EPR) spectroscopy (DPPH-EPR) test and Oxygen Radical Absorbance Capacity (ORAC) assay. The inhibition of lipid peroxidation and the level of inflammatory markers have been also investigated using lipopolysaccharide (LPS)-stimulated RAW 264 cells. It was revealed that ADE standardized to 25% of anthocyanins depresses the level of markers of inflammation and lipid peroxidation (Interleukin 1 beta (IL-1β), tumor necrosis factor alpha (TNF-α), and malondialdehyde (MDA)) in in vitro conditions. Additionally, it was confirmed that ADE at all analyzed concentrations did not show any cytotoxic effect as demonstrated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.

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Evaluation of antioxidant activity of Melothria maderaspatana in vitro

Open Life Sciences ◽

10.2478/s11535-010-0005-5 ◽

2010 ◽

Vol 5 (2) ◽

pp. 224-230 ◽

Cited By ~ 7

Author(s):

Boobalan Raja ◽

Kodukkur Pugalendi

Keyword(s):

Hydrogen Peroxide ◽

Antioxidant Activity ◽

Free Radical ◽

Hydroxyl Radical ◽

Superoxide Anion ◽

Radical Scavenging ◽

Reducing Power ◽

Free Radical Scavenging ◽

Melothria Maderaspatana

AbstractIn this study, an aqueous extract of leaves from Melothria maderaspatana was tested for in vitro antioxidant activity. Free radical scavenging assays, such as hydroxyl radical, hydrogen peroxide, superoxide anion radical and 2,2-diphenyl-1-picryl hydrazyl (DPPH), 2,2’-azinobis-(3-ethyl-enzothiazoline-6-sulfonic acid) (ABTS) radical scavenging, and reducing power assay, were studied. The extract effectively scavenged hydroxyl radical, hydrogen peroxide and superoxide anion radicals. It also scavenged DPPH and ABTS radicals. Furthermore, it was found to have reducing power. All concentrations of leaf extract exhibited free radical scavenging and antioxidant power, and the preventive effects were in a dose-dependent manner. The antioxidant activities of the above were compared to standard antioxidants such as butylated hydroxytoluene (BHT), ascorbic acid, and α-tocopherol. The results obtained in the present study indicate that the M. maderaspatana extract could be considered a potential source of natural antioxidant.

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CHEMICAL COMPOSITION AND HEPATOPROTECTIVE EFFECTS OF POLYPHENOLIC FRACTION FROM RIVEA HYPOCRATERIFORMIS IN PARACETAMOL INDUCED LIVER DAMAGE IN WISTAR ALBINO RATS

International Journal of Pharmacy and Pharmaceutical Sciences ◽

10.22159/ijpps.2016v8i10.13606 ◽

2016 ◽

Vol 8 (10) ◽

pp. 228

Author(s):

Shivakumar S. Godipurge ◽

Naveen J. Biradar ◽

Jaiprakash S. Biradar ◽

Nitin Mahurkar

Keyword(s):

Lipid Peroxidation ◽

Chemical Composition ◽

Hydroxyl Radical ◽

Liver Damage ◽

Radical Scavenging Activity ◽

Radical Scavenging ◽

Scavenging Activity ◽

Aerial Parts ◽

Hepatoprotective Effects ◽

Phenolic And Flavonoid

Objective: This study was designed to chemical composition and hepatoprotective effects of a polyphenolic fraction of aerial parts of R. hypocrateriformis (PPFRH). It was shown to exhibit strong in vitro lipid peroxidation and scavenging activity against hydroxyl radical.Methods: The chemical composition of a polyphenolic fraction of R. hypocrateriformis was analyzed by High-performance liquid chromatography method. Hepatocellular injuries induced by paracetamol were assessed by liver damage in Wistar albino rat; the hepatoprotective effect was evaluated by biochemical parameters in rat serum, antioxidant hydroxyl radical scavenging activity and lipid peroxidation in liver tissue.Results: The polyphenolic fraction of aerial parts of R. hypocrateriformis for lipid peroxidation is significantly (p<0.05). In the hepatoprotective activity of liver enzymes and hepatic necrosis were significantly (p<0.001) closer to paracetamol. The correlation coefficient between the hydroxyl scavenging radical and total phenolic and flavonoid contents were found to be R² = 0.9045 and R² = 0.8876 suggesting the contribution of phenolic and flavonoid compounds of the polyphenolic fraction of aerial parts of R. hypocrateriformis by 90% and 88% to its radical scavenging activity.Conclusion: The polyphenolic fraction of aerial parts of R. hypocrateriformis possesses a significant protective effect against acute hepatotoxicity induced by paracetamol and which may be due to the phenolic and flavonoid components.

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