• 대한약침학회지
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• 제24권1호
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• pp.24-31
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• 2021

Objectives: Accumulation of cellular reactive oxygen species (ROS) leads to oxidative stress. Increased production of ROS, such as superoxide anion, or a deficiency in their clearance by antioxidant defences, mediates cellular pathology. Grewia Spp fruits are a source of bioactive compounds and have notable antioxidant activity. Although the antioxidant capacity of Grewia Spp has been studied, there is very limited evidence that links the antioxidant activities of Grewia bicolor and Grewia flava to the inhibition of free radical formation associated with damage in biological systems. Methods: This study evaluated the protective effects of Grewia bicolor and Grewia flava extracts against free radical-induced oxidative stress and the resulting cytotoxicity effect using HeLa cells. Antioxidant properties determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and total phenolic content (TPC) assays showed significantly higher (p < 0.05) antioxidant activity in Grewia flava (ethanol extract) than Grewia flava (water extract) and Grewia bicolor (ethanol and water extracts). Results: Using 3-(4,5-dimethylthiazol-2-yl)-2,5diphenyltetrazolium bromide or MTT assay, cytotoxicity results showed that extracts of Grewia bicolor and Grewia flava were less toxic to HeLa cells at tested concentrations compared to the untreated control. This confirmed the low toxicity of these edible fruits at the tested concentrations in HeLa cells. Furthermore, hydrogen peroxide (H2O2)-induced cell loss was effectively reduced by pre-incubating HeLa cells with Grewia bicolor and Grewia flava extracts, with Grewia flava (ethanol extract) revealing better protection. Conclusion: The effect was speculated to be associated with the higher antioxidant activity of Grewia flava (ethanol extract). Additional studies will warrant confirmation of the mechanism of action of such effects.

• 한국식품과학회지
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• 제49권6호
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• pp.659-667
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• 2017

본 연구는 in vitro에서 나린진과 나린제닌의 항산화 활성을 평가하고, in vivo에서 BALB/c 마우스에 나린진과 나린제닌을 7일 동안 경구투여 한 후 6Gy의 감마선을 조사시켜 24시간 뒤 감마선 조사로 인한 세포 손상 및 산화적 스트레스에 대한 방사선 예방효과를 검토하였다. 나린진과 나린제닌의 항산화 활성평가에서 나린제닌이 나린진보다 ABTS 제거 활성 및 FRAP을 유의적으로 증가시켜 아글리콘 형태인 나린제닌이 글리코사이드 형태의 나린진보다 항산화 능력이 뛰어난 것을 확인하였다. 감마선 조사 전 나린제닌을 투여 한 군이 감마선 조사군에 비해 비장 지수, 흉선 지수 및 백혈구 수치가 증가하여 조혈 면역계 손상에 대해 보호효과가 있음을 확인하였다. 나린제닌을 투여 후 감마선을 조사한 군의 ALT와 AST가 감마선 조사군에 비해 유의적으로 감소하여 간세포 손상에 대한 예방효과를 확인하였다. 감마선 조사 전에 나린제닌을 투여한 군의 지방질과산화는 감마선 조사군에 비해 유의적으로 낮게 나타남을 보였으며, XO는 감마선 조사군에 비해 유의적으로 감소하여 방사선에 의한 장해를 감소시킨 것으로 판단할 수 있다. 감마선 조사 전 나린제닌을 투여한 군의 GSH와 항산화 효소의 활성은 감마선 조사군에 비해 유의적으로 증가하여 생체 내에 항산화 활성을 회복시켰다. 이 상의 결과를 통해 방사선 조사 전에 나린제닌의 투여는 방사선에 의한 세포 손상과 산화적 스트레스에 대해 보호 효과가 있어 방사선보호제로 유용하게 사용될 수 있다.

• Journal of Ginseng Research
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• 제34권3호
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• pp.198-204
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• 2010

Korean ginseng (Panax ginseng C.A. Meyer) preserved in syrup, known as ginseng Jung Kwa (GJK), is a popular traditional snack in Korea. We investigated the effects of heat processing time on total phenolic content and antioxidant capacity of GJK. Water extract was prepared from GJK with different heat processing times, 3 hours (GJK-3), 6 hours (GJK-6), or 12 hours (GJK-12), with sonication for 2 hours. The GJK extract contained total phenolic content in the following order: GJK-12 (2.28%)>GJK-6 (1.57%)>GJK-3 (1.29%). Both the peroxy and hydroxyl radical-scavenging activity and cellular antioxidant capacity of GJK extract was significantly enhanced with increasing heat processing time. The hydroxyl radical-scavenging activity of GJK-12 extract was greater than that of the GJK-3 and GJK-6 extracts, consistent with metal chelating capacity and reducing capacity. In a cellular model, the GJK extract effectively reduced 2,2'-azobis(2-amidinopropane) dihydrochloride, $Cu^{2+}$-, and $H_2O_2$-induced oxidative stress, with GJK-12 and GJK-6 extracts demonstrating greater cellular antioxidant capacity than the GJK-3 extract. These results suggest that heat processing time can contribute to the antioxidant capacity of GJK and that GJK extract may have the potential to be used as an effective dietary antioxidant to prevent oxidative stress-related diseases.

• Journal of Ginseng Research
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• 제41권2호
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• pp.169-179
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• 2017

Background: Ginsenosides are the main pharmacological components of Panax ginseng root, which are thought to be primarily responsible for the suppressing effect on oxidative stress. Methods: 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and oxygen radical absorption capacity were applied to evaluate the antioxidant activities of the ginsenosides. Human embryonic kidney 293 (HEK-293) cells were incubated with ginsenosides extracted by pulsed electric field (PEF) and solvent cold soak extraction (SCSE) for 24 h and then the injury was induced by $40{\mu}M$ $H_2O_2$. The cell viability and surface morphology of HEK-293 cells were studied using MTS assay and scanning electron microscopy, respectively. Dichloro-dihydro-fluorescein diacetate fluorescent probe assay was used to measure the level of intracellular reactive oxygen species. The intracellular antioxidant activities of ginsenosides were evaluated by cellular antioxidant activity assay in HepG2 cells. Results: The PEF extracts displayed the higher 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and stronger oxygen radical absorption capacity (with an oxygen radical absorption capacity value of $14.48{\pm}4.04{\mu}M\;TE\;per\;{\mu}g/mL$). The HEK-293 cell model also suggested that the protective effect of PEF extracts was dose-dependently greater than SCSE extracts. Dichloro-dihydro-fluorescein diacetate assay further proved that PEF extracts are more active (8% higher than SCSE extracts) in reducing intracellular reactive oxygen species accumulation. In addition, scanning electron microscopy images showed that the HEK-293 cells, which were treated with PEF extracts, maintained more intact surface morphology. Cellular antioxidant activity values indicated that ginsenosides extracted by PEF had stronger cellular antioxidant activity than SCSE ginsenosides extracts. Conclusion: The present study demonstrated the antioxidative effect of ginsenosides extracted by PEF in vitro. Furthermore, rather than SCSE, PEF may be more useful as an alternative extraction technique for the extraction of ginsenosides with enhanced antioxidant activity.

• KSBB Journal
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• 제29권6호
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• pp.432-436
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• 2014

We investigated antioxidative activity of the ethanol extracts of leaves of Cornus walteri Wanger (CWE) by treated enzyme in human dermal fibroblast (HDFs) irradiated by UVB. We examined the in vitro chemical and cellular antioxidant activities of CWE in HDFs. We employed scavenging assay for the 1,1-diphenyl-2,5-picrylhydrazyl (DPPH) radicals and cellular antioxidative activity of CWE, and we was investigated in $H_2O_2$-treated or UVB-irradiated HDFs. The CWE effectively scavenged DPPH radicals ($IC_{50}$ $7.03{\pm}0.4{\mu}g/mL$) when compared to the scavenging activities of L-ascorbic acid ($IC_{50}$ $4.69{\pm}0.3{\mu}g/mL$). CWE reduced UVB-induced cellular damage in HS68 cells by MTT assay and inhibited intracellular ROS generation in dose-dependent manner. In addition, CWE also attenuated the elevated levels of 8-isoprostane resulting from UVB-mediated oxidative stress. Collectively, these results suggest that CWE could be a new potential candidate as antioxidant against UVB-induced oxidative stress in HDFs.

• 한국미생물·생명공학회지
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• 제38권1호
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• pp.24-31
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• 2010

본 연구는 제주 연안 해수로부터 분리된 항산화물질을 생산하는 해양 방선균의 천연해양항산화물질에 관한 연구이다. 균주 ACT-18은 16S rDNA 염기서열, 전자현미경을 이용한 형태학적 분석, 생리학적, 생화학적 특성 및 세포 지방산분석을 통하여 동정되었다. 분리균주 ACT-18은 Gram 양성, 호기성, 비운동성 포자, 배지상에서 substrate mycelia의 색이 어두운 초록색, aerial mycelia는 노란색의 회색을 나타내였다. 세포사이즈는 약 $0.5{\sim}1.0\;{\mu}m$였다. 16S rDNA 염기서열 분석결과 Gram 양성박테리아인 Streptomyces 종들과 그룹을 형성하였다. 지방산분석결과 주요 지방산은 $C_{15:0}$ anteiso(39.33%), $C_{16:1}$ cis 9(11.96%), $C_{16:0}$ (13.08%)과 $C_{17:0}$ anteiso(10.99%)로 분석되었다. Streptomyce sp. ACT-18의 배양액 메탄올 추출물의 항산화 활성은 electron spin resonance (ESR) spectrometer에 의한 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl, alkyl 라디컬 소거활성을 측정하여 나타내었다. 그 결과 배양액 메탄올 추출물의 DPPH 라디컬 소거활성은 0.1 mg/mL의 농도에서 46%의 라디컬 소거활성을 보였으며, Hydroxyl 라디컬 소거활성은 0.1 mg/mL에서 63%, alkyl 라디컬 소거활성은 0.1 mg/mL에서 39%의 소거 활성을 보였다.

• Molecules and Cells
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• 제28권5호
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• pp.479-487
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• 2009

Glutathione reductase (GR) is an enzyme that recycles a key cellular antioxidant molecule glutathione (GSH) from its oxidized form (GSSG) thus maintaining cellular redox homeostasis. A recombinant plasmid to overexpress a GR of Brassica rapa subsp. pekinensis (BrGR) in E. coli BL21 (DE3) was constructed using an expression vector pKM260. Expression of the introduced gene was confirmed by semi-quantitative RT-PCR, immunoblotting and enzyme assays. Purification of the BrGR protein was performed by IMAC method and indicated that the BrGR was a dimmer. The BrGR required NADPH as a cofactor and specific activity was approximately 458 U. The BrGR-expressing E. coli cells showed increased GR activity and tolerance to $H_2O_2$, menadione, and heavy metal ($CdCl_2$, $ZnCl_2$ and $AlCl_2$)-mediated growth inhibition. The ectopic expression of BrGR provoked the co-regulation of a variety of antioxidant enzymes including catalase, superoxide dismutase, glutathione peroxidase, and glucose-6-phosphate dehydrogenase. Consequently, the transformed cells showed decreased hydroperoxide levels when exposed to stressful conditions. A proteomic analysis demonstrated the higher level of induction of proteins involved in glycolysis, detoxification/oxidative stress response, protein folding, transport/binding proteins, cell envelope/porins, and protein translation and modification when exposed to $H_2O_2$ stress. Taken together, these results indicate that the plant GR protein is functional in a cooperative way in the E. coli system to protect cells against oxidative stress.

• Animal Bioscience
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• 제34권3_spc호
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• pp.345-353
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• 2021

Phytobiotics, also known as phytochemicals or phytogenics, have a wide variety of biological activities and have recently emerged as alternatives to synthetic antibiotic growth promoters. Numerous studies have reported the growth-promoting effects of phytobiotics in chickens, but their precise mechanism of action is yet to be elucidated. Phytobiotics are traditionally known for their antioxidant activity. However, extensive investigations have shown that these compounds also have anti-inflammatory, antimicrobial, and transcription-modulating effects. Phytobiotics are non-nutritive constituents, and their bioavailability is low. Nonetheless, their beneficial effects have been observed in several tissues or organs. The health benefits of the ingestion of phytobiotics are attributed to their antioxidant activity. However, several studies have revealed that not all these benefits could be explained by the antioxidant effects alone. In this review, I focused on the bioavailability of phytobiotics and the possible mechanisms underlying their overall effects on intestinal barrier functions, inflammatory status, gut microbiota, systemic inflammation, and metabolism, rather than the specific effects of each compound. I also discuss the possible mechanisms by which phytobiotics contribute to growth promotion in chickens.

• BMB Reports
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• 제50권10호
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• pp.528-533
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• 2017

Peroxiredoxin I (Prx I) plays an important role as a reactive oxygen species (ROS) scavenger in protecting and maintaining cellular homeostasis; however, the underlying mechanisms are not well understood. Here, we identified a critical role of Prx I in protecting cells against ROS-mediated cellular senescence by suppression of $p16^{INK4a}$ expression. Compared to wild-type mouse embryonic fibroblasts (WT-MEFs), Prx $I^{-/-}$ MEFs exhibited senescence-associated phenotypes. Moreover, the aged Prx $I^{-/-}$ mice showed an increased number of cells with senescence associated-${\beta}$-galactosidase (SA-${\beta}$-gal) activity in a variety of tissues. Increased ROS levels and SA-${\beta}$-gal activity, and reduction of chemical antioxidant in Prx $I^{-/-}$ MEF further supported an essential role of Prx I peroxidase activity in cellular senescence that is mediated by oxidative stress. The up-regulation of $p16^{INK4a}$ expression in Prx $I^{-/-}$ and suppression by overexpression of Prx I indicate that Prx I possibly modulate cellular senescence through $ROS/p16^{INK4a}$ pathway.

• International Journal of Oral Biology
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• 제30권3호
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• pp.77-84
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• 2005

EGCG[(-)-epigallocatechin gallate], is a major component of green tea has been considered as a major antioxidant constituent. It has been considered as potential chemopreventive and chemotherapeutic agents. However, very little is known about the cellular actions by which EGCG mediates its therapeutic effects. Various aspects of antioxidant activity of EGCG were evaluated in this study. EGCG itself did not show significant cytotoxicity. Significant 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity was observed in all ranges of concentration ($0.8-100{\mu}g/ml$) used in this study. Protective effect of EGCG against hydrogen peroxide induced cell death was observed. Relatively high lipid peroxidation inhibitory activity were detected ($IC_{50}$ was about $20{\mu}g/ml$). EGCG also dose-dependently enhanced the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) in V79-4 cells. In concentrations of $100{\mu}g/ml$ of EGCG, activities of SOD, CAT and GPX were measured as 36.9 U/mg of protein, 22.9 U/mg of protein and 17.8 U/mg of protein, respectively. When these values were compared with those of the control groups (24.9 U/mg of protein, 14.9 U/mg of protein and 11.7 U/mg of protein), the relative increases were calculated as 48, 54 and 52%, respectively. Taken together, our findings suggest that EGCG can act as an antioxidant by scavenging radicals and enhancing antioxidant enzyme activities.