• Journal of Microbiology and Biotechnology
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• v.29 no.4
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• pp.518-526
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• 2019

Pueraria montana var. lobata is a bioactive substance with various beneficial health effects and has long been extensively used as a traditional medication for the treatment of fever, acute dysentery, diabetes, and cardiovascular diseases in Northeast Asian countries. The purpose of this study was to evaluate the cytoprotective activity of Pueraria montana var. lobata ethanol extract (PLE) for ultraviolet B (UVB)-induced oxidative stress in human dermal fibroblasts (HDF). It was hypothesized that PLE treatment ($25-100{\mu}g/ml$) would reduce intracellular reactive oxygen species (ROS) levels as well as increase collagen production in UVB-irradiated HDF. The results confirmed this theory, with collagen production increasing in the PLE treatment group in a dose-dependent manner. In addition, regulators of cellular ROS accumulation, including HO-1 and NOQ-1, were activated by Nrf2, which was mediated by PLE. Hence, intracellular levels of ROS were also reduced in the PLE treatment group in a dose-dependent manner. In conclusion, PLE increases collagen production and maintains hyaluronic acid (HA) levels in human dermal fibroblasts exposed to UVB-irradiation, thereby inhibiting photoaging.

• Clinical and Experimental Reproductive Medicine
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• v.49 no.2
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• pp.110-116
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• 2022

Objective: Sperm vitrification leads to the production of reactive oxygen species (ROS) that can damage the functional parameters of sperm. The present study aimed to investigate the antioxidant effect of Nigella sativa extract on motility, plasma membrane function, mitochondrial membrane potential (MMP), DNA damage, and intracellular ROS production. Methods: A total of 20 sperm samples were used. Samples were divided into six experimental groups, including groups with aqueous extract from N. sativa seeds at concentrations of 1% to 6%, a cryopreserved control group, and a fresh control group. Results: Statistical analysis showed significantly higher total sperm motility at concentrations of 3% to 6% than in the vitrified semen control group. Additionally, progressive motility and all motion characteristics at all concentrations were significantly higher than in the vitrified semen control group. The presence of N. sativa seed extract also improved the quality of the sperm parameters assayed in all experimental groups (1%-6%; intracellular ROS production, DNA damage, MMP, and sperm membrane function) compared to the control group. Conclusion: Higher concentrations of N. sativa led to improvements in all sperm parameters and sperm quality. These findings indicate that N. sativa seed extract is effective for improving the quality of sperm after vitrification.

• Korean Journal of Agricultural Science
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• v.49 no.2
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• pp.259-267
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• 2022

Although pesticides are recognized as necessary substances to improve agricultural production, exposure to pesticides is known to have a direct or indirect adverse effect on the reproductive function of mammals. The present study examines the effects of mancozeb, a well-known fungicide, on the fertility capacity of spermatozoa. Boar spermatozoa exposed to varying concentrations of mancozeb (0.01 - 0.5 µM) were evaluated for motility, motion kinetic parameters, viability, acrosome integrity and the generation of intracellular reactive oxygen species (ROS) after 30 min or 2 hrs of incubation. A significant reduction in the motility of spermatozoa was observed upon exposure to mancozeb. Similarly, there was a significant reduction of the motion kinematics of sperm treated with mancozeb as compared to untreated controls (p < 0.05). The sperm viability percentage and acrosome integrity also showed dose-dependent decreases upon exposure to mancozeb. High concentrations of mancozeb (0.2 - 0.5 µM) induced higher levels of intracellular ROS production, which resulted in the loss of the sperm membrane and decreased sperm motility due to oxidative stress. Taken together, the results here indicate that direct exposure to mancozeb affects the sperm fertility capacity. Hence, careful research that examines the interaction between reproduction and environmental toxins is crucial to prevent fertility disorders in animals.

• Development and Reproduction
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• v.27 no.2
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• pp.77-89
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• 2023

Metformin is the most widely used anti-diabetic drug that helps maintain normal blood glucose levels primarily by suppressing hepatic gluconeogenesis in type II diabetic patients. We previously found that metformin induces apoptotic death in H4IIE rat hepatocellular carcinoma cells. Despite its anti-diabetic roles, the effect of metformin on hepatic de novo lipogenesis (DNL) remains unclear. We investigated the effect of metformin on hepatic DNL and apoptotic cell death in H4IIE cells. Metformin treatment stimulated glucose consumption, lactate production, intracellular fat accumulation, and the expressions of lipogenic proteins. It also stimulated apoptosis but reduced autophagic responses. These metformin-induced changes were clearly reversed by compound C, an inhibitor of AMP-activated protein kinase (AMPK). Interestingly, metformin massively increased the production of reactive oxygen species (ROS), which was completely blocked by compound C. Metformin also stimulated the phosphorylation of p38 mitogen-activated protein kinase (p38MAPK). Finally, inhibition of p38MAPK mimicked the effects of compound C, and suppressed the metformin-induced fat accumulation and apoptosis. Taken together, metformin stimulates dysregulated glucose metabolism, intracellular fat accumulation, and apoptosis. Our findings suggest that metformin induces excessive glucose-induced DNL, oxidative stress by ROS generation, activation of AMPK and p38MAPK, suppression of autophagy, and ultimately apoptosis.

• Journal of Microbiology and Biotechnology
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• v.26 no.2
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• pp.277-286
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• 2016

Makgeolli lees (ML) has several physiological effects such as antioxidant, antidiabetic, and anticancer properties, but its biological functions have not been determined definitively. Here, we tested whether ML has a cytoprotective effect on paraquat (PQ)-induced oxidative stress in the human lung carcinoma cell line A549. At 0.1 mg/ml ML, viability of PQ-exposed A549 cells was restored by 12.4%, 18.5%, and 48.6% after 24, 48, and 72 h, respectively. ML also reduced production of the intracellular reactive oxygen species (ROS) that were generated by PQ treatment. Further experiments revealed that ML treatment enhanced the expression and nuclear translocation of nuclear factor erythroid 2-related factor 2 (NRF2) as well as ARE-GFP reporter activity. ML treatment also effectively increased the expression of NRF2's target genes NAD(P)H dehydrogenase quinone 1 (NQO1) and heme oxygenase 1 (HO-1). Moreover, we found that expression of cytoprotective genes, including glutathione peroxidases (GPXs), superoxide dismutase (SOD1), catalase (CAT), peroxiredoxin 3 (PRDX3), and peroxiredoxin 4 (PRDX4), was greatly enhanced by treatment with ML during PQ exposure. Taken together, the data suggest that treatment of PQ-exposed A549 cells with ML ameliorates cytotoxicity through induction of NRF2 expression and its target genes HO-1, NQO1, and other antioxidant genes. Thus, ML may serve as a functional food applicable to ROS-mediated human diseases.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.6
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• pp.907-915
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• 2014

Alpha-lipoic acid (${\alpha}$-LA) is not only involved in energy metabolism, but is also a powerful antioxidant that can protect against hepatic oxidative stress induced by some drugs, toxins, or under various physiological and pathophysiological conditions. Here, we investigated the effect of ${\alpha}$-LA against liver oxidative damage in broilers exposed to aflatoxin $B_1$ ($AFB_1$). Birds were randomly divided into four groups and assigned different diets: basal diet, 300 mg/kg ${\alpha}$-LA supplementation in basal diet, diet containing 74 ${\mu}g/kg$ $AFB_1$, and 300 mg/kg ${\alpha}$-LA supplementation in diet containing 74 ${\mu}g/kg$ $AFB_1$, for 3 weeks. The results revealed that the addition of 300 mg/kg ${\alpha}$-LA protected against the liver function damage of broilers induced by chronic low dose of $AFB_1$ as estimated by a significant (p<0.05) change in levels of plasma total protein, albumin, alkaline phosphatase and the activities of liver glutamic-oxalacetic transaminase and glutamic-pyruvic transaminase. The histopathological analysis also showed that liver tissues were injured in the $AFB_1$ diet, but this effect was alleviated by the addition of 300 mg/kg ${\alpha}$-LA. Additionally, $AFB_1$ induced a profound elevation of oxidative stress in birds, as indicated by an increase in malondialdehyde level, a decrease in glutathione peroxidase activity and a depletion of the glutathione content in the liver. All of these negative effects were inhibited by treatment with ${\alpha}$-LA. Our results suggest that the inhibition of $AFB_1$-induced excess production of lipid peroxides and the maintenance of intracellular antioxidant status may play important roles in the protective effects of ${\alpha}$-LA against $AFB_1$-induced oxidative damage in the liver.

• Journal of Embryo Transfer
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• v.32 no.4
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• pp.257-265
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• 2017

Ferulic Acid (FA) is a metabolite of phenylalanine and tyrosine, a phenolic compound commonly found in fruits and vegetables. Several studies have shown that FA has various functions such as antioxidant effect, prevention of cell damage from irradiation, protection from cell damage caused by oxygen deficiency, anti-inflammatory action, anti-aging action, liver protective effect and anti-cancer action. In this study, we investigated the maturation rate, intracellular glutathione (GSH) and reactive oxygen species (ROS) of porcine oocytes by adding FA to the in vitro maturation (IVM) medium and examined subsequent embryonic developmental competence at 5% oxygen through parthenogenesis. There is no significant difference between the control group ($0{\mu}M$) and treatment groups ($5{\mu}M$, $10{\mu}M$, $20{\mu}M$) on maturation rates. Intracellular GSH levels in oocyte treated with $5{\mu}M$ of FA significantly increased (P < 0.05), and $20{\mu}M$ of FA revealed significant decrease (P < 0.05) in intracellular ROS levels compared with the control group. Oocytes treated with FA exhibited significantly higher cleavage rates (79.01% vs 89.19%, 92.20%, 90.89%, respectively) than the control group. Oocytes treated with $10{\mu}M$ showed significantly higher blastocyst formation rates (28.3% vs 40.3%, respectively) after PA than the control group. Total cell numbers in blastocyst of $10{\mu}M$ FA displayed significantly higher (39.4 vs 51.9, respectively) than the control group. In conclusion, these results suggested that treatment with FA during IVM improved the developmental potential of porcine embryos by increasing intracellular GSH synthesis and reducing ROS levels. Also, there was an improvement of cleavage rate, blastocyst formation and total cell numbers in blastocysts. It might be associated with Keap1-Nrf2 pathway as an antioxidant regulate pathway that plays a crucial role in determining the sensitivity of cells to oxidative damages by regulating the basal and inducible expression of enzymes which is related to detoxification and anti-oxidative effects, stress response enzymes and/or proteins and ABC transporters.

• Food Science and Biotechnology
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• v.17 no.5
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• pp.1086-1091
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• 2008

The antioxidant activities of the extracts of dangyuja (Citrus grandis Osbeck) leaves were evaluated. The highest phenolic content was obtained from the ethyl acetate fraction (EF) (202.1$\pm$0.8 mg GAE/g dried extract) and it exhibited the highest 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity. The cytoprotective effects of EF on oxidative damage induced by tert-butyl hydroperoxide (t-BHP) in a human hepatoma cell line, HepG2 cells, were investigated to understand the intracellular antioxidant mechanisms. Treatment of HepG2 cells with EF prior to oxidative stress was found to inhibit reactive oxygen species (ROS) generation, lipid peroxidation, and DNA damage in a dose-dependent manner. Gas chromatography-mass spectrometry (GC-MS) studies on EF resulted in tentative identification of 19 compounds representing 94.3% of the total content. Taken together, these results demonstrated that EF has excellent antioxidant activities and thus dangyuja leaves have great potential as a source for natural antioxidant which can be applied in food products.

• Korean Journal of Pharmacognosy
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• v.36 no.1 s.140
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• pp.34-37
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• 2005

Reactive oxygen species (ROS) are known to cause oxidative modification of DNA, proteins, lipids and small cellular molecules and are associated with tissue damage and are the contributing factors for inflammation, aging, cancer, arteriosclerosis, hypertension and diabetes. We screened the anti-oxidants in V79-4 hamster lung fibroblast cells induced by hydrogen peroxide with eighteen pure compounds isolated from natural products. Allantoin, brassicasterol, and hypaconitine were found to strongly scavenge intracellular reactive oxygen species, which is measured by dichlorodihydrofluorescin diacetate method (DCHF-DA), and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical.

• International Journal of Industrial Entomology and Biomaterials
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• v.45 no.1
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• pp.1-11
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• 2022

Silkworm pupae, a by-product of the silk industry are known to be valuable resource of nutrients for humans as well as animals besides encompassing diverse bioactive constituents. However, there is a paucity of knowledge on their role in amelioration of oxidative stress and anti-skin aging properties. In the present study, we evaluated the inhibitory effect of aqueous as well as ethanolic (30% and 70%) extracts from distinct stages of male and female silkworm pupae belonging to two silkworm varieties on skin aging-related enzymes. The activities of collagenase, elastase and tyrosinase were effectively inhibited by 70% ethanolic silkworm pupal extracts (SPE), followed by 30% with aqueous extracts exhibiting meager inhibitory potential. SPE were also investigated for their antioxidant activity in oxidative-stressed murine fibroblasts (L929). The intracellular ROS and lipid peroxidation induced by tert-butyl hydroperoxide (t-BHP) in fibroblasts was better attenuated by pre treatment with ethanolic (30%) and aqueous extracts, respectively. The safety of the extracts was determined by studying their effect on fibroblast cell viability and it was found that none of the extracts were cytotoxic. Our findings indicate the potential utility of SPE as anti-aging components in cosmeceuticals.