• 생명과학회지
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• 제29권8호
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• pp.888-894
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• 2019

류마티스관절염(rheumatoid arthritis)과 골관절염(osteoarthritis) 같은 관절질환은 연골세포(chondrocytes) 감소와 관절연골(articular cartilage)의 분해를 수반한다. 최근, 연골세포의 활성과 연골 항상성(cartilage homeostasis)에 염증성 ROS (reactive oxygen species) burst 및 나이와 관련된 산화적 스트레스(oxidative stress)의 증가와 관련된 연구가 활발히 진행되고 있다. 본 연구는 관절연골로부터 분리한 연골세포의 노화 과정과 나이대별 관절연골에서 항산화 인자들(antioxidants)의 변화를 조사함으로써, 연골세포와 관절연골의 노화 과정 동안 산화적 스트레스로부터 조직을 보호하는 항산화 인자들의 역할을 규명하는데 목적이 있다. 쥐의 관절연골로부터 분리한 연골세포의 연속 계대배양을 통한 노화 과정에서 산화적 스트레스가 증가함을 관찰하였다. 그리고, 노화 유도한 연골세포는 세포내 총 glutathione (GSSG/GSH) 양과 항산화 효소 superoxide dismutase (SOD)와 heme oxygenase-1 (HO-1)의 발현이 증가하였다. 다음으로, 나이대별 쥐로부터 분리한 관절연골에서 항산화 인자의 발현을 분석하였다. 항산화 인자 glutathione의 양은 40주령에서 발현이 가장 높게 관찰되었으며 72주령에 다소 감소하였고, SOD와 HO-1의 발현은 나이대별로 현저히 증가되는 경향을 보였다. 이를 종합해 볼 때, 세포내 항산화 인자들은 과도한 양의 ROS에 반응하여 연골세포의 노화와 나이와 관련된 관절연골의 퇴화 과정에서 중요한 역할을 하는 것으로 보인다.

• 한국식품과학회지
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• 제46권5호
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• pp.601-608
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• 2014

본 연구에서는 녹색 및 자색 콜라비 착즙액의 이화학적 특성(pH 및 $^{\circ}Bx$), 총 페놀함량, 항산화 활성(ORAC 지수, DPPH radical 소거능, ABTS radical 소거능, FRAP 및 reducing power), 아질산염 소거능, 피부 섬유아세포에서의 세포 보호 효과 및 ROS 생성억제 효과를 측정하였다. 총 페놀 함량은 녹색 콜라비 착즙액(153.74 mg GAE/mL)에서 자색 콜라비 착즙액(122.55 mg GAE/mL)보다 더 높게 나타난 반면, ORAC 지수의 경우 자색 콜라비 착즙액($664.32{\mu}MTE/mL$ )에서 녹색 콜라비 착즙액($572.74{\mu}MTE/mL$)보다 더 높게 나타났다. DPPH radical 소거능, ABTS radical 소거능, FRAP 및 reducing power에서는 콜라비 착즙액 10, 50, 100% 농도에서 농도 의존적으로 항산화 활성이 증가하는 경향을 나타내었으며, 같은 농도로 비교하였을 때, 녹색 콜라비 착즙액에서 자색 콜라비 착즙액보다 높은 항산화 활성을 보였다. 아질산염 소거능에서도 녹색 콜라비 착즙액이 자색 콜라비 착즙액보다 높은 효능을 나타내었다. 피부 섬유아세포에서는 녹색 및 자색 콜라비 착즙액을 동결 건조하여 사용하였으며 10, 25, $50{\mu}g/mL$의 농도에서 독성을 나타내지 않았다. Hydrogen peroxide로 산화적 스트레스를 유도한 상태에서 세포 보호 효과를 측정한 결과 $10{\mu}g/mL$의 농도에서는 녹색 및 자색 콜라비 착즙액에서 hydrogen peroxide를 처리한 군과 차이를 나타내지 않았으나, 25, $50{\mu}g/mL$의 농도에서는 농도 유의적으로 세포보효효과가 증가하였다. 또한 녹색 콜라비 착즙액에서는 양성대조군으로 사용한 항산화 물질인 NAC 수준까지 세포생존율이 증가하였다. $H_2$-DCFDA 염색을 통하여 관찰한 ROS 생성 억제 효과는 세포 보호 효과와 유사한 경향으로 관찰되었다. 이상의 결과를 종합하여 볼 때 녹색 및 자색 콜라비 착즙액이 다양한 항산화 모델에서 효능을 나타내었으며, 피부 섬유아세포에서 세포 보호 효과 및 ROS 생성 억제효과가 관찰되어 기능성 식품원료로서의 활용도가 매우 넓을 것으로 판단된다.

• Animal Bioscience
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• 제36권5호
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• pp.710-719
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• 2023

Objective: The present study investigated whether protodioscin (PD), a steroidal saponin mainly found in rhizome of Dioscorea species, alleviates oxidative stress-induced damage of porcine oocytes during in vitro maturation. Methods: Oocytes were treated with different concentrations of PD (0, 1, 10, 100, and 200 µM) in the presence of 200 µM H₂O₂ during in vitro maturation. Following maturation, spindle morphology and mitogen-activated protein kinase activity was assessed along with reactive oxygen species level, GSH activity, and mRNA expression of endogenous antioxidant genes at the MII stage. On the day 7 after parthenogenetic activation, blastocyst formation rate was calculated and the quality of embryo and mRNA expression of development-related genes was evaluated. Results: Developmental competence was significantly poorer in the 0 µM PD-treated (control) group than in the non-treated (normal) and 10 µM PD-treated (10PD) groups. Although the reactive oxygen species level did not significantly differ between these three groups, the glutathione level and mRNA expression of antioxidant genes (superoxide dismutase 1 [SOD1], SOD2, nuclear factor erythroid 2-related factor 2 [Nrf2], and hemo oxygenase-1 [HO-1]) were significantly higher in the normal and 10PD groups than in the control group. In addition, the percentage of oocytes with defective spindle and abnormal chromosomal alignment was significantly lower and the ratio of phosphorylated p44/42 to total p44/42 was significantly higher in the normal and 10PD groups than in the control group. The total cell number per blastocyst was significantly higher in the 10PD group than in the control group. The percentage of apoptotic cells in blastocysts was highest in the control group; however, the difference was not significant. mRNA expression of development-related genes (POU domain, class 5, transcription factor 1 [POU5F1], caudal type homeobox 2 [CDX2], Nanog homeobox [NANOG]) was consistently increased by addition of PD. Conclusion: The PD effectively improves the developmental competence and quality of blastocysts by protecting porcine oocytes against oxidative stress.

• 대한약침학회지
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• 제14권3호
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• pp.55-61
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• 2011

Objectives: Reactive Oxygen Species(ROS) are continuously produced at a high rate as a by-product of aerobic metabolism. Since tissue damage by free radical, ROS such as hydrogen peroxide($H_2O_2$), nitric oxide(NO) increases with age. Several lines of evidence provided that ROS appears to cause to develop aging-related various diseases such as cancer, arthritis, cardiovascular disease. In this study, we have conducted to investigate the pharmacological effects of red ginseng for the development possibility to pharmacopuncture drug sources or healthy aid foods. Methods: For our aims, it was investigated the biological activities of Red Ginseng ethanol extracts (RGEE) by measuring total polyphenol contents, total flavonoid contents, DPPH radical scavenging activity, ABTS radical scavenging activity and cell viability of MCF 10A and SK-MEL-2 in vitro with MTT assay method. Results: The total polyphenol contents of RGEE was 3.06${\pm}$0.11mg/g in 10mg/ml, the total flavonoid contents of RGEE was 1.35${\pm}$0.01mg/g in same concentration. The ABTS radical scavenging activity was about 80% and that of DPPH activity was 65% in 50mg/ml of RGEE. The cell viability of SKMEL-2, skin cancer cell line was decreased and that of MCF 10A, skin normal cell line was increased. Conclusions: We conclude that RGEE may be useful as potential functional foods or pharmacopuncture drug sources on the diseases induced by oxidant stress.

• Journal of Web Engineering
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• 제14권5호
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• pp.1044-1057
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• 2022

Helicobacter pylori (H. pylori) causes gastric diseases by increasing reactive oxygen species (ROS) and interleukin (IL)-8 expression in gastric epithelial cells. ROS and inflammatory responses are regulated by the activation of nuclear factor erythroid-2-related factor 2 (Nrf2) and the expression of Nrf2 target genes, superoxide dismutase (SOD) and heme oxygenase-1 (HO-1). We previously demonstrated that Korean red ginseng extract (RGE) decreases H. pylori-induced increases in ROS and monocyte chemoattractant protein 1 in gastric epithelial cells. We determined whether RGE suppresses the expression of IL-8 via Nrf2 activation and the expression of SOD and HO-1 in H. pylori-infected gastric epithelial AGS cells. H. pylori-infected cells were treated with RGE with or without ML385, an Nrf2 inhibitor, or zinc protoporphyrin (ZnPP), a HO-1 inhibitor. Levels of ROS and IL-8 expression; abundance of Keap1, HO-1, and SOD; levels of total, nuclear, and phosphorylated Nrf2; indices of mitochondrial dysfunction (reduction in mitochondrial membrane potential and ATP level); and SOD activity were determined. As a result, RGE disturbed Nrf2-Keap1 interactions and increased nuclear Nrf2 levels in uninfected cells. H. pylori infection decreased the protein levels of SOD-1 and HO-1, as well as SOD activity, which was reversed by RGE treatment. RGE reduced H. pylori-induced increases in ROS and IL-8 levels as well as mitochondrial dysfunction. ML385 or ZnPP reversed the inhibitory effect of RGE on the alterations caused by H. pylori. In conclusion, RGE suppressed IL-8 expression and mitochondrial dysfunction via Nrf2 activation, induction of SOD-1 and HO-1, and reduction of ROS in H. pylori-infected cells.

• 미생물학회지
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• 제51권4호
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• pp.319-328
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• 2015

Schizosaccharomyces pombe $sdu1^+$ 유전자는 PPPDE superfamily member에 속하는 탈유비퀴틴화 효소인 Sdu1을 엔코딩한다. 이전 연구에서, $sdu1^+$ 유전자 포함 재조합 플라스미드 pYSTP를 사용하여 Sdu1이 카르복시 말단 유비퀴틴 가수분해 활성을 보유한다는 사실이 입증된 바 있다. 본 연구에서는, 높은 배양 온도에 대한 Sdu1의 열내성적 역할이 검토되었다. 온도 천이 실험에서, pYSTP 포함 S. pombe 세포들이 37도나 42도로 천이한 후에 벡터 대조 세포들보다 훨씬 더 잘 성장하였다. 37도나 42도로 천이 한 후 6시간 배양한 pYSTP 포함 S. pombe 세포들이 벡터 대조 세포들보다 낮은 활성산소종 수준을 나타내었다. pYSTP 포함 S. pombe 세포들이 온도 천이와 관계없이 벡터 대조 세포들보다 다소 낮은 일산화질소 수준을 나타내었다. pYSTP 포함 S. pombe 세포들이 벡터 대조 세포들보다 훨씬 높은 총 글루타치온 수준을 나타내었다. 온도천이 후의 총 수퍼옥시드 디스무타아제 및 글루타치온 과산화효소 활성들이 pYSTP 포함 S. pombe 세포들에서 더 높은 것으로 측정되었다. 요약하면, S. pombe Sdu1은 활성산소종과 일산화질소 수준은 낮추고, 총 글루타치온, 총 수퍼옥시드 디스무타아제 및 글루타치온 과산화효소 수준들은 증가시킴으로써 높은 배양 온도에 대한 열내성적 역할을 나타낸다.

• 한국자원식물학회지
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• 제19권6호
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• pp.649-654
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• 2006

The objective of present study was to investigate the anti oxidative and hepatoprotective effects of tomato extracts. Total antioxidant capacity and total antioxidant response were 5.5 and $19.8{\mu}g$ Trolox equivalent per mg of tomato extract, respectively. DPPH radical scavenging activity of tomato extracts ($10mg\;ml^{-1}$) was 70% as compared to 100% by pyrogallol solution as a reference. The effect of the tomato extracts on lipid peroxidation was examined using rat liver mitochondria induced by iron/ascorbate. Tomato extracts at the concentration of $0.5mg\;ml^{-1}$ significantly decreased TBARS concentration. Tomato extracts prevented lipid peroxidation in a dose-dependent manner. The effect of the tomato extracts on reactive oxygen species (ROS) generation was examined using cell-free system induced by $H_2O_2/FeSO_4$. Addition of $1mg\;ml^{-1}$ of tomato extracts significantly reduced dichlorofluorescein (DCF) fluorescence. Tomato extracts caused concentration-dependent attenuation of the increase in DCF fluorescence, indicating that tomato extracts significantly prevented ROS generation in vitro. The effect of tomato extracts on cell viability and proliferation was examined using hepatocyte culture. Primary cultures of rat hepatocytes were incubated with 1mM tert-butyl hydroperoxide (t-BHP) for 90 min in the presence or absence of tomato extracts. MTT values by addition of tomato extracts at the concentration of 2, 10, and $20mg\;ml^{-1}$ in the presence of t-BHP were 13, 33 and 48%, respectively, compared to 100% as control. Tomato extracts increased cell viability in a dose-dependent manner. These results demonstrate that tomato extracts suppressed lipid peroxidation and t-BHP-induced hepatotoxicity and scavenged ROS generation. Thus antioxidant and hepatoprotective effects of tomato extracts seem to be due to, at least in part, the prevention from free radicals-induced oxidation, followed by inhibition of lipid peroxidation.

• Journal of Ginseng Research
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• 제43권2호
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• pp.179-185
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• 2019

Background: Oxidative stress induces the production of reactive oxygen species (ROS), which play important causative roles in various pathological conditions. Black ginseng (BG), a type of steam-processed ginseng, has drawn significant attention due to its biological activity, and is more potent than white ginseng (WG) or red ginseng (RG). Methods: We evaluated the protective effects of BG extract (BGE) against oxidative stress-induced cellular damage, in comparison with WG extract (WGE) and RG extract (RGE) in a cell culture model. Ethanolic extracts of WG, RG, and BG were used to evaluate ginsenoside profiles, total polyphenols, flavonoid contents, and antioxidant activity. Using AML-12 cells treated with $H_2O_2$, the protective effects of WGE, RGE, and BGE on cellular redox status, DNA, protein, lipid damage, and apoptosis levels were investigated. Results: BGE exhibited significantly enhanced antioxidant potential, as well as total flavonoid and polyphenol contents. ATP levels were significantly higher in BGE-treated cells than in control; ROS generation and glutathione disulfide levels were lower but glutathione (GSH) and NADPH levels were higher in BGE-treated cells than in other groups. Pretreatment with BGE inhibited apoptosis and therefore protected cells from oxidative stress-induced cellular damage, probably through ROS scavenging. Conclusion: Collectively, our results demonstrate that BGE protects AML-12 cells from oxidative stress-induced cellular damages more effectively than WGE or RGE, through ROS scavenging, maintenance of redox status, and activation of the antioxidant defense system.

• BMB Reports
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• 제34권6호
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• pp.544-550
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• 2001

In pre-transplant total-body irradiation (TBI), the lung is a critical dose-limiting organ. Also, the possible role of oxidative stress was suggested in the development of TBI-induced lung damage. This study explores the association between TBI-induced oxidative stress and the induction of lung pathogenesis by investigating TBI-induced oxidative stress in the lungs of male C57BL/6 mice after a single dose of 10 Gy TBI. We showed significant increases of reactive oxygen species (ROS) formation and lipid peroxidation, and also a depletion and oxidation of glutathione after TBI. There is evidence that pretreatment with 1,10-phenanthroline (o-phen) significantly reduces oxidative stress in the lung. This indicates that the TBI-induced ROS generation involves a metal-catalyzed Fenton-type reaction. A pretreatment of buthionine sulfoximine (BSO) augmented the glutathione depletion and oxidation, but had no effect on the ROS formation and lipid peroxidation up to 6 h after TBI. Histopathological features that are consistent with pneumonitis were observed in the BSO pretreated-mice 1 week after irradiation. The results suggest that TBI-induced oxidative stress in the lung involves a generation of ROS through a Fenton-type reaction. Also, glutathione plays an important inhibitory role in the radiation-induced lung pathogenesis by participating in the self-amplifying cascade subsequent to the ROS generation by irradiation.

• 동아시아식생활학회지
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• 제24권3호
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• pp.359-367
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• 2014

This study analyzed phytochemicals, including various carotenoids, tocopherol and L-ascorbic acid, in green, yellow and orange paprikas (GP, YP and OP) and measured the preventive effects of lipophilic extracts from different colored paprikas on the blockage of gap junctional intercellular communication (GJIC), which is known as a cellular event associated with tumor promotion. Main carotenoids were lutein and ${\beta}$-carotene in GP, lutein, ${\beta}$-carotene, capsanthin, violaxanthin, ${\beta}$-carotene and capsorubin in YP, and lutein, ${\beta}$-carotene, cryptoxanthin and zeaxanthin in OP. Total carotenoid contents were $65.54{\pm}15.87$ mg/100 g dw in OP, $11.98{\pm}0.69$ mg/100 g dw in YP and $10.30{\pm}1.43$ mg/100 g dw in GP. Tocopherol contents were highest in GP compared with in YP and OP, whereas L-ascorbic acid contents were very high in all paprikas. We determined the non-cytotoxic levels of paprika extracts by MTT assay, which showed less formation of reactive oxygen species (ROS) induced by $500{\mu}M$ $H_2O_2$ for 1h. Finally, we showed that pretreatment of paprika extracts prevented inhibition of GJIC induced by $500{\mu}M$ $H_2O_2$ by the scrape-loading/dye-transfer technique. In conclusion, each colored paprika has unique phytochemicals and showed a protective effect on inhibition of GJIC.