• 약학회지
• /
• 제51권1호
• /
• pp.75-82
• /
• 2007

Reactive oxygen species (ROS) are produced in the metabolic process of oxygen in cells. The superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) in cells systemize the antioxidant enzymes to control the oxidative stress. Genistein is one of the isoflavonoids, and its role in controlling cellular oxidative stress is presently the active issue at question. In this study; we analyzed genistein-induced survival rates of the CHO-K1 cells, activities of antioxidant enzymes, ROS levels, and expression levels of antioxidant enzyme genes in order to investigate the effect of genistein on cellular ROS production and antioxidative systems in CHO-K1 cells. As results, the survival rate of cells was decreased as the dose of genistein increases (12.5${\sim}$200 ${\mu}$M). Genistein increased cellular ROS levels, while it reduced total SOD activities and the expression of CuZnSOD. In conclusion, we suggest that genistein may induce oxidative stress via down-regulation of SOD.

• 약학회지
• /
• 제49권1호
• /
• pp.86-91
• /
• 2005

Flavonoids are class of polyphenolic compounds widely distributed in the plant kingdom, which display a variety of biological activities, including antiviral, antithrombotic, antiinflammatory, antihistaminic, antioxidant and free-radica 1 scavenging abilities. The antioxidant enzyme (AOE) system plays an important role in the defense against oxidative stress insults. To determine whether flavonoid, myricetin can exert antioxidative effects not only directly by modulating the AOE system but also scavenging free radical, we investigated the influence of the flavonoid myricetin on cell viability, different antioxidant enzyme activities, ROS level and the expression of different antioxidant emzyme in B16F10 murine melanoma cells. Myricetin in a concentration range from 6.25 to $50\;{\mu}M$ decreased superoxide dismutase (SOD) and glutathione peroxidase (GPx) enzyme activities, but catalase (CAT) activity was increased. In the myricetin-treated group, ROS levels were decreased dose-dependently. Antioxidant enzyme expression was measured by RT-PCR. Myricetin treatment of B16F10 cells increased catalase expression. Expression levels of copper zinc superoxide dismutase (CuZn SOD) were not affected by exposure of myricetin. Manganese superoxide dismutase (Mn SOD) and GPx expression levels decreased slightly after myricetin treatment. In conclusion, the antioxidant capacity of myricetin was due to CAT and free-radical scavenging.

• Molecular & Cellular Toxicology
• /
• 제5권4호
• /
• pp.273-282
• /
• 2009

Cadmium (Cd) and tributyltin (TBT) are common contaminants of marine and freshwater ecosystems, and can induce the formation of reactive oxygen species (ROS). These ROS can, in turn, cause oxidative stress. In the present study, we investigated time-related effects of Cd (0.05 and 0.1 ppm) and TBT (5 and 10 ppb) treatment on antioxidant enzyme activity, i.e., the activity of superoxide dismutase (SOD) and catalase (CAT) in the gills and digestive glands of the ark shell, Scapharca broughtonii. In addition, hydrogen peroxide ($H_2O_2$) concentrations, lysozyme activity, and glutamate oxaloacetate transaminase (GOT) and glutamate pyruvate transaminase (GPT) levels were measured in the hemolymph. We found that Cd and TBT treatment significantly increased antioxidant enzyme mRNA expression and activity in the digestive glands and gills in a time-dependent manner. In response to the Cd and TBT treatments, antioxidant enzymes mRNA expression and activity increased up to day 5 in the digestive glands and then decreased by day 7. In the gills, antioxidant enzymes mRNA expression and activity increased up to day 3 and then decreased by day 5. Likewise, $H_2O_2$ concentrations significantly increased up to day 5 and then decreased by day 7. Finally, lysozyme activity decreased during the experimental period, whereas GOT and GPT levels were significantly increased in a time-dependent manner. These results suggest that antioxidant enzymes play an important role in decreasing ROS levels and oxidative stress in ark shells exposed to Cd and TBT.

• Biomolecules & Therapeutics
• /
• 제24권1호
• /
• pp.33-39
• /
• 2016

Oxidative stress activates several intracellular signaling cascades that may have deleterious effects on neuronal cell survival. Thus, controlling oxidative stress has been suggested as an important strategy for prevention and/or treatment of neurodegenerative diseases. In this study, we found that ginsenoside Rh1 inhibited hydrogen peroxide-induced reactive oxygen species generation and subsequent cell death in rat primary astrocytes. Rh1 increased the expression of phase II antioxidant enzymes, such as heme oxygenase-1 (HO-1), NAD(P)H:quinone oxidoreductase 1, superoxide dismutase-2, and catalase, that are under the control of Nrf2/ARE signaling pathways. Further mechanistic studies showed that Rh1 increased the nuclear translocation and DNA binding of Nrf2 and c-Jun to the antioxidant response element (ARE), and increased the ARE-mediated transcription activities in rat primary astrocytes. Analysis of signaling pathways revealed that MAP kinases are important in HO-1 expression, and act by modulating ARE-mediated transcriptional activity. Therefore, the upregulation of antioxidant enzymes by Rh1 may provide preventive therapeutic potential for various neurodegenerative diseases that are associated with oxidative stress.

• 대한한방내과학회지
• /
• 제31권1호
• /
• pp.25-39
• /
• 2010

Background : At high glucose levels in $\beta$-cells, cell viability and insulin secretion are decreased by glucotoxicity. Sopyung-tang(SPT) had an effect on blood glucose level decrease and antioxidant enzyme activities in streptozotocin-induced diabetic rats. Objectives : This study performed a series of experiment to verify the effects of SPT extract on the cell viability, antioxidant enzyme activities, insulin secretion and insulin mRNA expression at hyperglycemic states of RIN-m5F. Methods : After treatment at various concentrations of SPT added to the RIN-m5F cells, cell viability by MTT assay, free radical-scavenging activity, SOD activity and insulin secretion were measured. Additionally, insulin-related gene expression was measured using real-time RT-PCR. Results : Compared to the control group, SPT extract showed considerable effects on RIN-m5F cell viability, DPPH radical-scavenging activity, superoxide dismutase (SOD) activity, insulin secretion and insulin-related gene expression. Conclusions : This study showed that SPT extract has an effect on $\beta$-cell cell viability, insulin secretion and insulin-related gene expression. Thus, SPT extract may be used for treatment of diabetes and its complications. Further mechanism studies of SPT seem to be necessary on the glucotoxicity and oxidative stress.

• Nutrition Research and Practice
• /
• 제8권3호
• /
• pp.272-277
• /
• 2014

BACKGROUND/OBJECTIVES: This study investigated the antioxidant activities and hepatoprotective effects of Schisandra chinensis Baillon extract (SCE) against tert-butyl hydroperoxide (t-BHP)-induced oxidative hepatic damage in rats. MATERIALS/METHODS: Sprague-Dawley (SD) rats were pretreated with SCE (300, 600, and 1,200 mg/kg BW) or saline once daily for 14 consecutive days. On day 14, each animal, except those belonging to the normal control group, were injected with t-BHP (0.8 mmol/kg BW/i.p.), and all of the rats were sacrificed 16 h after t-BHP injection. RESULTS: Although no significant differences in AST and ALT levels were observed among the TC and SCE groups, the high-dose SCE group showed a decreasing tendency compared to the TC group. However, erythrocyte SOD activity showed a significant increase in the low-dose SCE group compared with the TC group. On the other hand, no significant differences in hepatic total glutathione (GSH) level, glutathione reductase (GR), and glutathione peroxidase (GSH-Px) activities were observed among the TC and SCE groups. Hepatic histopathological evaluation revealed that pretreatment with SCE resulted in reduced t-BHP-induced incidence of lesions, such as neutrophil infiltration, swelling of liver cells, and necrosis. In particular, treatment with a high dose of SCE resulted in induction of phase II antioxidant/detoxifying enzyme expression, such as glutathione S-transferase (GST) and glutamate-cysteine ligase catalytic subunit (GCLC). CONCLUSIONS: Based on these results, we conclude that SCE exerts protective effects against t-BHP induced oxidative hepatic damage through the reduction of neutrophil infiltration, swelling of liver cells, and necrosis. In addition, SCE regulates the gene expression of phase II antioxidant/detoxifying enzymes independent of hepatic antioxidant enzyme activity.

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

• The Korean Journal of Physiology and Pharmacology
• /
• 제23권6호
• /
• pp.519-528
• /
• 2019

Mitochondrial dysfunction is closely associated with reactive oxygen species (ROS) generation and oxidative stress in cells. On the other hand, modulation of the cellular antioxidant defense system by changes in the mitochondrial DNA (mtDNA) content is largely unknown. To determine the relationship between the cellular mtDNA content and defense system against oxidative stress, this study examined a set of myoblasts containing a depleted or reverted mtDNA content. A change in the cellular mtDNA content modulated the expression of antioxidant enzymes in myoblasts. In particular, the expression and activity of glutathione peroxidase (GPx) and catalase were inversely correlated with the mtDNA content in myoblasts. The depletion of mtDNA decreased both the reduced glutathione (GSH) and oxidized glutathione (GSSG) slightly, whereas the cellular redox status, as assessed by the GSH/GSSG ratio, was similar to that of the control. Interestingly, the steady-state level of the intracellular ROS, which depends on the reciprocal actions between ROS generation and detoxification, was reduced significantly and the lethality induced by $H_2O_2$ was alleviated by mtDNA depletion in myoblasts. Therefore, these results suggest that the ROS homeostasis and antioxidant enzymes are modulated by the cellular mtDNA content and that the increased expression and activity of GPx and catalase through the depletion of mtDNA are closely associated with an alleviation of the oxidative stress in myoblasts.

• 동의생리병리학회지
• /
• 제25권3호
• /
• pp.451-458
• /
• 2011

The medicinal mushroom Inonotus obliquus is a traditional and widely used multi-functional fungus. In this study, we have investigated whether Inonotus obliquus (Chaga mushroom) extracts exerts anti-oxidant effects on cisplatin-induced cytotoxicity in auditory cell line, HEI-OC1 cells. First of all, Chaga extracts has no harmful effects on viability of HEI-OC1 cells in the dose range of $65{\sim}125{\mu}g/m{\ell}$. Moreover, it shows cyto-protective effects on the cells treated with cisplatin-induced cytotoxicity in HEI-OC1 cells and the damage of hair cells arrays of the rat primary organ of Corti explants in the presence of cisplatin. Pretreatment with Chaga extracts inhibited the cell death, reactive oxygen species generation (ROS), lipid peroxidation induced by cisplatin. These effects were associated with the induction of antioxidant enzyme by Chaga extracts. We further investigated the effects of Chaga extracts on expression of antioxidant enzymes such as Cu, Zn superoxide dismutase (SOD 1) and Mn SOD (SOD 2) by RT-PCR. In addition, Chaga extracts shows SOD activity and SOD protein expression in cisplatin treated group induced similar to control group. Taken together, these results indicate that Chaga extracts can prevent cisplatin-induced cytotoxicity by radical-scavenging activity (SOD activity) in HEI-OC1 cells. It might be an effective as antioxidant and further studies on the chemo-preventive mechanisms of Inonotus obliquus are needed.

• 생명과학회지
• /
• 제23권6호
• /
• pp.743-750
• /
• 2013

본 연구의 목적은 혈관계 순환을 촉진시키는 약효제를 발굴하는 것이다. 아사이베리(Acai berry) 에탄올 추출물(ABEE)의 항산화 효과 뿐만 아니라 토끼의 폐에서 유래한 angiotensin converting enzyme (ACE)의 활성 및 대식세포와 혈관내피세포에서 nitric oxide 생성에 대한 효과가 본 연구에서 조사되었다. 먼저 환원력과 지질과산화에 대한 ABEE의 항산화효과가 관찰되었다. 뿐만 아니라, ABEE는 hydroxyl radical에 의하여 유발된 DNA 산화에 대한 보호효과를 보여주었다. 더욱이 ABEE는 0.01%의 농도에서 약 50% 정도의 angiotensin converting enzyme 활성에 대한 억제효과를 발휘하는 것으로 나타났다. 혈관내피세포에서 ABEE는 nitric oxide 의 생성을 증가시켰으나 대식세포에서는 lipopolysaccharide에 의하여 생성된 nitric oxide 생성을 감소시켰다. 게다가 superoxide dismutase (SOD)-2와 -3 발현수준은 ABEE 처리에 의하여 증가되었으나 SOD-1의 발현수준은 일정하였다. 더욱이 nitric oxide synthases-1 (NOS-1)의 발현수준은 ABEE 처리에 의하여 증가되었으나, 유발 효소인 NOS-2의 발현수준은 일정하였다. 또한 SOD의 전사인자인 Nrf-2의 발현수준은 ABEE에 의하여 증가되었다. 그러므로 이러한 결과들은 ABEE가 이상의 작용을 경유하여 혈액순환을 촉진시킬 수 있고, 혈관의 건강을 위한 큰 잠재성을 가지고 있다는 것을 암시하고 있다.