• Biomolecules & Therapeutics
• /
• 제4권4호
• /
• pp.402-407
• /
• 1996

Effects of oxidative stress on the induction of apoptosis and the activity of antioxidant enzymes were investigated in HL-60 cells using $H_2O$$_2$and cisplatin which generate oxygen species in the cell. Various concentrations of oxidants were treated to cells and at different incubation time, cells were harvested for assays. Cell viability, morphology by propidium iodide staining and DNA fragmentation by agarose gel electrophoresis were observed to determine whether they induce apoptosis. The activity of antioxidant enzymes such as superoxide dismutase and catalase was also measured to evaluate the cellular response to the oxidative damage. The results are as follows: $H_2O$$_2$ induced apoptosis at 10 $\mu$M after 6h incubation, while it took 12h for cisplatin. Both oxidants induced the superoxide dismutase activity at a tolerable low concentration. However, at a concentration which causes apoptotic cell death, the enzyme level was dropped markedly at first and then recovered to the normal level after which it declined again, probably due to cell death. On the other hand, changes in the activity of catalase were not significant at most concentrations except the statistically significant decrease at 24h after 10 $\mu$M-$H_2O$$_2$treatment. In this study, $H_2O$$_2$- and cisplatintreated cells showed similar results in apoptotic response and enzyme activities, suggesting that anticancer activity of cisplatin may be related, at least in part, to the production of oxygen free radicals.

• Natural Product Sciences
• /
• 제18권2호
• /
• pp.76-82
• /
• 2012

Stamens of Mesua ferrea L. are a well-known herbal drug used in Indian System of Traditional Medicine to treat various diseases. The claimed activity of this plant part is necessitated to investigate antioxidant and hepatoprotective activity. Authenticated plant sample was extracted with hexane, ethanol (EtOH) and water (aq.) using ASE 100 accelerated solvent extractor. Antioxidant activity was evaluated by means of different in vitro assays. Hepatoprotective effect was investigated on carbon tetrachloride induced oxidative stress in liver slice culture model. Cytotoxic marker lactate dehydrogenase (LDH) released in culture medium and the activity of lipid peroxidation along with antioxidant enzymes (AOEs) namely superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR) were estimated. Hexane and EtOH extracts were significantly inhibited DPPH, NO, SOD and $ABTS^+$ radical in dose dependent manner. The trade of phenol content was: aq. extract < hexane extract < EtOH extract. A significant correlation was shown by total phenol content and free radical scavenging activity of extracts. The culture system treated with hexane extract, EtOH extract or ascorbic acid exhibited significant depletion in LDH, lipid peroxidation, antioxidative enzymes SOD, CAT and GR. Hexane extract and EtOH extracts of stamen of M. ferrea protected liver slice culture cells by alleviating oxidative stress induced damage to liver cells.

• BMB Reports
• /
• 제40권6호
• /
• pp.1039-1049
• /
• 2007

Overdoses of acetaminophen cause hepato-renal oxidative stress. The present study was undertaken to investigate the protective effect of a 43 kDa protein isolated from the herb Cajanus indicus, against acetaminophen-induced hepatic and renal toxicity. Male albino mice were treated with the protein for 4 days (intraperitoneally, 2 mg/kg body wt) prior or post to oral administration of acetaminophen (300 mg/kg body wt) for 2 days. Levels of different marker enzymes (namely, glutamate pyruvate transaminase and alkaline phosphatase), creatinine and blood urea nitrogen were measured in the experimental sera. Intracellular reactive oxygen species production and total antioxidant activity were also determined from acetaminophen and protein treated hepatocytes. Indices of different antioxidant enzymes (namely, superoxide dismutase, catalase, glutathione-S-transferase) as well as lipid peroxidation end-products and glutathione were determined in both liver and kidney homogenates. In addition, Cytochrome P450 activity was also measured from liver microsomes. Finally, histopathological studies were performed from liver sections of control, acetaminophen-treated and protein pre- and post-treated (along with acetaminophen) mice. Administration of acetaminophen increased all the serum markers and creatinine levels in mice sera along with the enhancement of hepatic and renal lipid peroxidation. Besides, application of acetaminophen to hepatocytes increased reactive oxygen species production and reduced the total antioxidant activity of the treated hepatocytes. It also reduced the levels of antioxidant enzymes and cellular reserves of glutathione in liver and kidney. In addition, acetaminophen enhanced the cytochrome P450 activity of liver microsomes. Treatment with the protein significantly reversed these changes to almost normal. Apart from these, histopathological changes also revealed the protective nature of the protein against acetaminophen induced necrotic damage of the liver tissues. Results suggest that the protein protects hepatic and renal tissues against oxidative damages and could be used as an effective protector against acetaminophen induced hepato-nephrotoxicity.

• 한국식품영양과학회지
• /
• 제28권2호
• /
• pp.471-476
• /
• 1999

The effects of total dietary restriction(100% restriction of energy intake) on thiobarbituric acid reactive substance(TBARS) contents and intracellular antioxidant enzymes activities in the liver and kidney of young male Sprague Dawley rats were studied. The TBARS contents were reduced in both liver and kidney, up to 77% and 79% of the control rats, fed ad libitum, respectively at 7 days after dietary restriction . Superoxide dismutase(SOD) activities in the liver and kidney of rats were increased significantly by total dietary restriction. However, the activity of catalase in kidney was decreased 27% at 6 days after dietary restriction, but this enzyme activity did not change in liver. The changes of glutathione peroxidae(GSHPx) and catalase activities in the liver and kidney of rats with dietary restriction were not significant. These result suggested that dietary restriction reduce the free radical induced by tissue damage, as determined by TBARS content, in both the liver and kidney but the changes of activities of antioxidant enzymes may not be a contributory factor in reducing oxidative damage to tissue.

• 한국환경성돌연변이발암원학회지
• /
• 제25권4호
• /
• pp.129-133
• /
• 2005

We investigated the cytoprotective effect of phloroglucinol, which was isolated from Ecklonia cava (brown seaweed), against oxidative stress induced cell damage in Chinese hamster lung fibroblast(V79-4) cells. Phloroglucinol was found to scavenge intracellular reactive oxygen species (ROS) generated by $\gamma-ray$ radiation. In addition, Phloroglucinol inhibited cell damage induced by radiation through scavenging ROS. Phloroglucinol increased the superoxide dismutase and glutathione peroxidase activity, Taken together, the results suggest that phloroglucinol protectes V79-4 cells against oxidative damage by enhancing the cellular antioxidant enzymes activity.

• 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.

• 환경생물
• /
• 제20권2호
• /
• pp.173-179
• /
• 2002

N-nitrosoethylurea(NEU)에 의한 지질 과산화물의 함량 변화와 알데히드 대사효소 및 항산화효소의 활성변화를 쥐 간세포에서 측정하였다. 알데히드 대사효소로는 alcohol dehydrogenase와 aldehyde dehydrogenase가 사용되었고 항산화효소로는 glutathione transferase, superoxide dismutase, glutathione reductase와 catalase가 사용되었다. 쥐 간세포에 다양한 농도의 NEU를 처리한 후 지질 과산화물의 함량변화를 측정하였다. 그 결과 6.25mM NEU에 의하여 지질 과산화물의 함량이 최대 2.5배 증가하였다. Alcohol dehydrogenase의 활성은 NEU처리에 의하여 대조군보다 최대 2.3배 증가하였고 aldehyde dehydrogenase의 활성은 약 2배 증가하였다. 전암성 병변의 지표로 이용되는 glutathione transferase와 catalase의 경우 NEU처리에 의한 활성증가가 미미하였다. 그러나 superoxide dismutase의 활성은 최대 1.5배 증가하였고, glutathione reductase의 활성은 약 3배 증가하였다. 그러므로 superoxide dismutase와 g1utathione reductase의 활성증가가 NEU의 독성에 대한 세포내 항산화 방어과정에서 중요한 역할을 할 것으로 추측된다

• Preventive Nutrition and Food Science
• /
• 제16권1호
• /
• pp.12-17
• /
• 2011

This study was designed to investigate the protective effect of Sasa borealis leaf extract on 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative stress in LLC-PK1 cells (porcine kidney epithelial cells). The butanol fraction from Sasa borealis leaf extract (SBBF) was used in this study because it possessed strong antioxidant activity and high yield among fractions. Exposure of LLC-PK1 cells to 1 mM AAPH for 24 hr resulted in a significant decrease in cell viability, but SBBF treatment protected LLC-PK1 cells from AAPH-induced cell damage in a dose dependant manner. To determine the protective action of SBBF against AAPH-induced damage of LLC-PK1 cells, we measured the effects of SBBF on lipid peroxidation and antioxidant enzymes activities of AAPH treated cells as well as scavenging activities on superoxide anion radical and hydroxyl radical. SBBF had a protective effect against the AAPH-induced LLC-PK1 cellular damage and decreased lipid peroxidation and increased activities of antioxidant enzymes such as superoxide dismutase and glutathione peroxidase. Furthermore, SBBF showed strong scavenging activity against superoxide anion radical. The $IC_{50}$ value of SBBF was $28.45{\pm}1.28\;{\mu}g/mL$ for superoxide anion radical scavenging activity. The SBBF also had high hydroxyl radical scavenging activity ($IC_{50}=31.09{\pm}3.08\;{\mu}g/mL$). These results indicate that SBBF protects AAPH-induced LLC-PK1 cells damage by inhibiting lipid peroxidation, increasing antioxidant enzyme activities and scavenging free radicals.

• Preventive Nutrition and Food Science
• /
• 제20권3호
• /
• pp.210-214
• /
• 2015

Grape pomace is an abundant source of underutilized winery by-products. Polyphenols were extracted from grape pomace using cellulase and gluco-amylase enzymes. 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Folin-Ciocalteu's assays were used to measure antioxidant activity and total polyphenolic contents. Both cellulase, and gluco-amylase digested grape pomace showed efficient radical scavenging activity. In addition, the total polyphenolic contents of cellulase digested grape pomace showed lower concentrations were effective compared to higher concentrations, whereas glucoamylase enzyme did not show remarkable variations. The DPPH radical scavenging activity and total polyphenolic contents were significantly higher in the cellulase digested grape pomace compared to the gluco-amylase digested and the not digested grape pomace. It is notable that enzymatic digestions were efficient for extracting polyphenols from grape pomace. The underutilized grape pomace polyphenols can be further used for food safety as a natural antioxidant.

• 한국응용과학기술학회지
• /
• 제36권3호
• /
• pp.985-994
• /
• 2019

뽕잎 (Morus alba L.)을 노루궁뎅이 버섯균사체(Hericium erinaceum)로 발효시켜 열수 추출한 발효물(MA-HE)이 항산화 효소인 Cu,Zn-superoxide dismutase (SOD)와 ceruloplasmin (CP)의 산화적 변형에 미치는 영향을 관찰하였다. 세포질에서 중요한 항산화 활성을 나타내는 Cu,Zn-SOD와 CP는 모두 구리이온을 포함하는 단백질이다. MA-HE은 peroxyl radical에 의해 변형되는 Cu,Zn-SOD와 CP를 모두 보호하였다. MA-HE은 peroxyl radical에 의한 Cu,Zn-SOD의 단백질 fragmentation과 효소의 inactivation을 모두 효과적으로 억제하였다. 또한, MA-HE는 CP의 fragmentation과 inactivation도 효과적으로 억제하였다. MA-HE가 peroxyl radical을 소거할 수 있는 기능을 가지고 있는 지를 확인하기 위해 thiobarbituric acid를 이용한 활성 측정법으로 관찰한 결과, MA-HE는 농도가 증가함에 따라 비례적으로 peroxyl radical 소거 활성이 높아지는 것으로 관찰되었다. MA-HE는 $100{\mu}g/mL$에서 44.03% 소거활성을 나타냈다. 따라서, MA-HE는 peroxyl radical을 효과적으로 소거함으로서 세포 내의 항산화 효소를 산화적 손상으로부터 보호할 것으로 생각된다. 이와 같은 결과는 MA-HE가 세포 내에서 발생하는 활성산소종을 효과적으로 소거하여 산화적 스트레스에 의해 야기되는 세포 독성에 대한 보호 작용을 할 것으로 생각된다.