scholarly journals Antioxidant Peptides from the Protein Hydrolysate of Monkfish (Lophius litulon) Muscle: Purification, Identification, and Cytoprotective Function on HepG2 Cells Damage by H2O2

Marine Drugs ◽  
2020 ◽  
Vol 18 (3) ◽  
pp. 153 ◽  
Author(s):  
Xiao-Meng Hu ◽  
Yu-Mei Wang ◽  
Yu-Qin Zhao ◽  
Chang-Feng Chi ◽  
Bin Wang

In the work, defatted muscle proteins of monkfish (Lophius litulon) were separately hydrolyzed by pepsin, trypsin, and in vitro gastrointestinal (GI) digestion methods, and antioxidant peptides were isolated from proteins hydrolysate of monkfish muscle using ultrafiltration and chromatography processes. The antioxidant activities of isolated peptides were evaluated using radical scavenging and lipid peroxidation assays and H2O2-induced model of HepG2 cells. In which, the cell viability, reactive oxygen species (ROS) content, and antioxidant enzymes and malondialdehyde (MDA) levels were measured for evaluating the protective extent on HepG2 cells damaged by H2O2. The results indicated that the hydrolysate (MPTH) prepared using in vitro GI digestion method showed the highest degree of hydrolysis (27.24 ± 1.57%) and scavenging activity on a 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical (44.54 ± 3.12%) and hydroxyl radical (41.32 ± 2.73%) at the concentration of 5 mg protein/mL among the three hydrolysates. Subsequently, thirteen antioxidant peptides (MMP-1 to MMP-13) were isolated from MPTH. According to their DPPH radical and hydroxyl radical scavenging activity, three peptides with the highest antioxidant activity were selected and identified as EDIVCW (MMP-4), MEPVW (MMP-7), and YWDAW (MMP-12) with molecular weights of 763.82, 660.75, and 739.75 Da, respectively. EDIVCW, MEPVW, and YWDAW showed high scavenging activities on DPPH radical (EC50 0.39, 0.62, and 0.51 mg/mL, respectively), hydroxyl radical (EC50 0.61, 0.38, and 0.32 mg/mL, respectively), and superoxide anion radical (EC50 0.76, 0.94, 0.48 mg/mL, respectively). EDIVCW and YWDAW showed equivalent inhibiting ability on lipid peroxidation with glutathione in the linoleic acid model system. Moreover, EDIVCW, MEPVW, and YWDAW had no cytotoxicity to HepG2 cells at the concentration of 100.0 µM and could concentration-dependently protect HepG2 cells from H2O2-induced oxidative damage through decreasing the levels of reactive oxygen species (ROS) and MDA and activating intracellular antioxidant enzymes of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px). These present results indicated that the protein hydrolysate and isolated antioxidant peptides from monkfish muscle, especially YWDAW could serve as powerful antioxidants applied in the treatment of some liver diseases and healthcare products associated with oxidative stress.

Marine Drugs ◽  
2019 ◽  
Vol 17 (9) ◽  
pp. 531 ◽  
Author(s):  
Zhao ◽  
Yang ◽  
Wang ◽  
Zhao ◽  
Chi ◽  
...  

For the full use of Spanish mackerel (Scomberomorous niphonius) muscle to produce antioxidant peptides, the proteins of Spanish mackerel muscle were separately hydrolyzed under five kinds of enzymes and in vitro gastrointestinal digestion, and antioxidant peptides were isolated from the protein hydrolysate using ultrafiltration and multiple chromatography methods. The results showed that the hydrolysate (SMPH) prepared using in vitro GI digestion showed the highest degree of hydrolysis (27.45 ± 1.76%) and DPPH radical scavenging activity (52.58 ± 2.68%) at the concentration of 10 mg protein/mL among the six protein hydrolysates, and 12 peptides (SMP-1 to SMP-12) were prepared from SMPH. Among them, SMP-3, SMP-7, SMP-10, and SMP-11 showed the higher DPPH radical scavenging activities and were identified as Pro-Glu-Leu-Asp-Trp (PELDW), Trp-Pro-Asp-His-Trp (WPDHW), and Phe-Gly-Tyr-Asp-Trp-Trp (FGYDWW), and Tyr-Leu-His-Phe-Trp (YLHFW), respectively. PELDW, WPDHW, FGYDWW, and YLHFW showed high scavenging activities on DPPH radical (EC50 1.53, 0.70, 0.53, and 0.97 mg/mL, respectively), hydroxyl radical (EC50 1.12, 0.38, 0.26, and 0.67 mg/mL, respectively), and superoxide anion radical (EC50 0.85, 0.49, 0.34, and 1.37 mg/mL, respectively). Moreover, PELDW, WPDHW, FGYDWW, and YLHFW could dose-dependently inhibit lipid peroxidation in the linoleic acid model system and protect plasmid DNA (pBR322DNA) against oxidative damage induced by H2O2 in the tested model systems. In addition, PELDW, WPDHW, FGYDWW, and YLHFW could retain their high activities when they were treated under a low temperature (<60 °C) and a moderate pH environment (pH 5–9). These present results indicate that the protein hydrolysate, fractions, and isolated peptides from Spanish mackerel muscle have strong antioxidant activity and might have the potential to be used in health food products.


INDIAN DRUGS ◽  
2014 ◽  
Vol 51 (10) ◽  
pp. 38-42
Author(s):  
N. K Choudhary ◽  
◽  
J Dwivedi ◽  
S Sharma

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.


2015 ◽  
Vol 2015 ◽  
pp. 1-8
Author(s):  
Xiao-Tian Zhang ◽  
Chun-Jiang Yu ◽  
Jian-Wei Liu ◽  
Yan-Ping Zhang ◽  
Chao Zhang ◽  
...  

We analyzed the effects of a traditional Chinese medicine, Qizhi Jiangtang Jiaonang (QJJ), on insulin resistance (IR) in vitro. After an in vitro model of IR was established by treating human liver cancer cells (HepG2 cells) with palmitic acid, the cells were then treated with various concentrations of QJJ. Treatment with 400 µM palmitic acid for 24 h induced IR in HepG2 cells. The survival rate for HepG2 cells in the IR group was significantly lower than that of the untreated control group (P< 0.001); however, QJJ restored HepG2 cell survival (P< 0.001). As compared with HepG2 cells in the IR group, QJJ at all doses analyzed significantly increased glucose consumption (allP< 0.05). Moreover, treatment with all the QJJ doses significantly reduced the mean intracellular reactive oxygen species levels as compared with the IR group (allP< 0.05). Furthermore, high-dose QJJ reduced both TNF-αand IL-6 levels as compared to the IR group (allP< 0.05). QJJ ameliorated the altered PI3K, GLUT4, and RAGE expression observed with IR. In conclusion, QJJ can improve IR in HepG2 cells, which may be mediated through the IRS-1/PI3K/GLUT4 signaling pathway as well as regulation of NF-κB-mediated inflammation and oxidative stress.


2011 ◽  
Vol 361-363 ◽  
pp. 748-752 ◽  
Author(s):  
Jiao Jiao Mu ◽  
Qian Cheng Zhao ◽  
Jian Wei Li

The Mercenaria mercenaria protein hydrolysates (MMPH) and Ruditapes philippinarum protein hydrolysates (RPPH) were prepared by three enzymatic methods, and the radical scavenging activity of the protein hydrolysates was analyzed. The results showed that the protein hydrolysates prepared by the combination of Protamex and Flavourzyme exhibited significant radical scavenging activity; For DPPH radical, the EC50values of MMPH and RPPH were 33.0 mg/mL and 93.0 mg/mL, respectively, and for hydroxyl radical, the EC50values of the protein hydrolysates were 7.5 mg/mL and 7.0 mg/mL, respectively; The protein hydrolysates were further fractionated by ultrafiltration membrane of 6000 Da, and the radical scavenging activity of the resultant fraction with lower molecular weight (<6000 Da) was significantly (P<0.05) increased, and for DPPH radical, the EC50values of MMPH and RPPH fractions were 29.5 mg/mL and 58.5 mg/mL, respectively, and for hydroxyl radical, the EC50 values of the fractions were 6.0 mg/mL and 7.0 mg/mL, respectively.


2015 ◽  
Vol 6 (6) ◽  
pp. 1919-1927 ◽  
Author(s):  
Fengchao Zha ◽  
Binbin Wei ◽  
Shengjun Chen ◽  
Shiyuan Dong ◽  
Mingyong Zeng ◽  
...  

A shrimp by-product protein hydrolysate via the Maillard reaction could alleviate cellular damage, but result in higher HMF and loss of nutritional quality.


Molecules ◽  
2020 ◽  
Vol 25 (18) ◽  
pp. 4114
Author(s):  
Jozef Fejér ◽  
Ivan Kron ◽  
Daniela Gruľová ◽  
Adriana Eliašová

In the present study, in vitro hydroxyl radical scavenging activities, total phenols and terpene contents in 70% ethanol extracts were evaluated. Samples of crushed (CB) and non-crushed ripe juniper berries (NCB) collected at five localities in North-East Slovakia during the years 2012–2014 were compared. Standard or modified methods for determining phytochemical and antioxidant activity were used together with a novel method for the evaluation of the results after the correction of the measured values per gram of dry matter (DM). Statistically significantly higher DM contents (average values for three years) were found in the CB extracts (ranging from 18.86 to 21.91 mg/mL) in comparison to those for NCB (ranging from 2.59 to 9.90 mg/mL). Depending on the localities and years, the hydroxyl radical scavenging activity ranged from 15.52 to 32.85% for NCB and from 65.59 to 88.12% for CB, respectively. The contents of total phenols ranged from 43.75 to 246.75 mg gallic acid equivalents (GAE)/L (NCB) and from 151.84 to 278.19 mg GAE/L (CB), respectively. However, the higher content of total phenols per gram of DM was found in the NCB extracts (8.49–42.23 mg GAE/g DM) and then in CB (6.87–18.77 GAE/g DM). The results obtained in this study showed a higher efficiency for extraction from juniper berries in 70% ethanol if the pericarp was disrupted in comparison to that achieved with the maceration of intact berries.


Author(s):  
Archana V ◽  
Indumathy R

Objective: The aim of this study is to evaluate the protective effect of Delonix elata (L.) leaf extract against doxorubicin-induced cardiotoxicity in H9c2 cells. Methods: Doxorubicin has been used to treat cancer, but its clinical uses are limited because of its dose-dependent cardiotoxicity. Reactive oxygen species play an important role in the pathological process of cardiotoxicity. The various extracts (pet.ether, ethyl acetate and ethanol) of Delonix elata leaves antioxidant property was evaluated by SOD antioxidant assay and DPPH free radical scavenging assay. The cells were incubated with different concentrations of various extracts of Delonix elata leaves for 2 hr, followed by incubation with 5µM doxorubicin for 24 hr. Cell viability was determined by using MTT assay, respectively. Results: The various extracts of Delonix elata leaves exhibits antioxidant activity. The Doxorubicin significantly decreased cell viability which was accompanied by an increased ROS production. Pre-treatment with various extracts of Delonix elata leaves increased the viability ofcells and inhibit the generation of reactive oxygen species. Conclusion: In this study, findings how that Delonix elata leaf extract exhibited a protective effect against oxidative stress-induced cardiomyocyte damage. The ethanolic extract of Delonix elata leaves possesses significant antioxidant and cardioprotective activity.


Luminescence ◽  
2007 ◽  
Vol 22 (4) ◽  
pp. 379-386 ◽  
Author(s):  
Irena Kruk ◽  
Hassan Y. Aboul-Enein ◽  
Teresa Michalska ◽  
Krzysztof Lichszteld ◽  
Katarzyna Kubasik-Kladna ◽  
...  

Author(s):  
Abdul Sadat ◽  
Mayukh Hore ◽  
Kaushik Chakraborty ◽  
Subhrajyoti Roy

Objective: The present study was carried out to evaluate the in vitro preliminary phytochemical analysis and antioxidant activity of methanolic leaf extracts of Corchorus olitorius L. The in vitro antioxidant activity was evaluated by DPPH radical scavenging activity method.Methods: Fresh jute leaves (immature, mature and senescence) were collected, air dried and the crude powder was prepared for phytochemical and antioxidant analysis. The powder was mixed with 70% methanol and the supernatant was separated. The antioxidant activity of this methanolic extract was measured on the basis of the scavenging activity of the stable 1, 1-diphenyl 2-picrylhydrazyl (DPPH) free radical with slight modifications. Phytochemical analysis was performed according to standard laboratory protocol.Results: The results indicated the presence of different phytochemicals viz. glycosides, steroids, cholesterol, alkaloids, phenols, flavonoids, riboflavin, saponins and terpenoids. The sample also showed antioxidant activity by inhibiting DPPH radical. The significant antioxidant activity of methanolic leaf extract of might be due to the presence of saponins, phenols, flavonoids and alkaloids found in the preliminary phytochemical analysis.Conclusion: Present study reveals that the jute leaf possesses different phytonutrients and exhibited DPPH radical scavenging activity, and therefore, may be used for therapeutic purposes.


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