• KSBB Journal
• /
• v.10 no.5
• /
• pp.561-567
• /
• 1995

Using superoxide dismutase (SOD)-deficient mutants of Saccharomyces cerevisiae, the oxygen toxicity induced by paraquat was studied. In aerobic culture condition, yeasts lacking MnSOD (milochondrial SOD) showed more significant growth retardation than CuZnSOD (cytoplasmic SOD)-deficient yeasts. However, not so big differences in growth pattern of those mutants compared with wild type were observed under anaerobic condition. When exposed to paraquat, the growth of yeasts lacking CuZnSOD was severely affected by higher than 0.01mM of paraquat in culture medium. By the analysis of several cellular components ivolved in free radical generating and scavenging system, it was found that, under aerobic condition, the content of lipid peroxides in cell membrane as well as cellular activity of glutathion peroxidase of CuZnSOD-deficient mutants was increased in the presence of paraquat, although significant decrease of catalase activity was observed in those stratns. In MnSOD-deficient yeast, however, increment in cellular activity of glutathion peroxldase and catalase by paraquat was observed without any deterioration of membrane lipid. It implies that the lack of mitochondrial SOD could be compensated by both of glutathion peroxldase and catalase, but that only glutathion peroxidase might act for CuZnSOD in cytoplasm. In contrast, all of SOD-deficient mutants showed a significant decrease in catalase activity, but slight increase in the activities of glutathion peroxidase, when cultivated anaerobically in the medium containing paraquat. Nevertheless, any significant changes of lipid peroxides in cell membranes were not observed during anaerobic cultivation of SOD-deficient mutants. It suggests that a little amount of free radicals generated by paraquat under anaerobic condition could be sufficiently overcome by glutathion peroxidase but not by catalase.

• Journal of Life Science
• /
• v.31 no.6
• /
• pp.559-567
• /
• 2021

This study compared the nutritional characteristics and antioxidant effects of sea mustards sourced from five different areas (Barammaegi, Gultongmeori, Chanmulgae, Johongtaek, and Goraedeung) in Taejongdae, Youngdo, Busan. The contents of total flavonoids and phenols and fatty acid composition were measured. To evaluate their antioxidant effects, 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays were used. Acetone/methylene chloride (A+M) extracts from all the sea mustards contained higher amounts of total flavonoids and phenols than methanol (MeOH) extracts. Among the sea mustards obtained from the different areas, the total flavonoid and total phenolic content of the A+M extract of the sea mustard from Gultongmeori was 1.44±0.04 mg/g and 1.72±0.06 mg/g, respectively. In terms of the fatty acid composition, the Gultongmeori sea mustard had higher percentages of total n-6, total n-3, eicosapentaenoic acid (EPA, 20:5n-3), and docosahexaenoic acid (DHA, 22:6n-3) than the sea mustards from the other areas. The A+M extract of the sea mustard from Gultongmeori was more effective in terms of scavenging free radicals as compared with that of the other sea mustards, as assessed by the DPPH and ABTS assays (p<0.05). In a 120-minute reactive oxygen species (ROS) production assay, all the extracts tested decreased cellular ROS production induced by H₂O₂ compared to that produced by exposure to an extract-free control (p<0.05). The extracts from Barammaegi and Gultongmeori had a greater inhibitory effect on cellular ROS production. These results indicated that the antioxidant effects of sea mustards might be associated with a higher amount of flavonoids and phenols. This study suggests that food-processed products from sea mustard can be developed as functional foods for promoting health in the local population.

• Tuberculosis and Respiratory Diseases
• /
• v.39 no.2
• /
• pp.120-130
• /
• 1992

Background: Although there have been many studies on the pathogenetic mechanism of acute lung injury, it is still elusive. Recently interests have been focused on the role of oxygen free radicals. But the effect of hydroxyl radical on the neutrophil mobilization and the alveolar-capillary permeability is not clear especially in the endotoxin-induced acute lung injury model of rats. This investigation was performed to evaluate the pathogenetic role of hydroxyl radical on the neutrophil accumulation into the lung and the increased alveolar-capillary permeability in the endotoxin-induced acute lung injury in rats. Method: Fifty rats were divided into four groups: vehicle control group (n=5, 6hrs; n=5, 24hrs), endotoxin-treated group (n=10, 6hrs; n=10, 24hrs), Dimethylthiourea (DMTU)-pretreated group (n=10, 6hrs), and deferoxamine (DFX)-pretreated group (n=10, 6hrs). Thirty minutes before sacrifice, $^{125}I$-tagged bovine serum albumin was injected. Six and twenty four hours after endotoxin injection, the rats were sacrificed, and the radioactivity of lung tissue and peripheral blood was counted. Permeability index was defined as the ratio of radioactivity between lung tissue and peripheral blood. Another set of rats (n=52) were divided into the same four groups as before [vehicle control group (n=5, 6hrs; n=5, 24hrs), endotoxin·treated group (n=7, 6hrs; n=8, 24hrs), DMTU-pretreated group (n=6, 6hrs; n=9, 24hrs), and DFX-pretreated group (n=5, 6hrs; n=7, 24hrs)], and were sacrificed 6 and 24 hours after endotoxin injection. In these rats, cell profile of peripheral blood and bronchoalveolar lavage fluid was evaluated, and the pathologic examination of lung tissue was performed. Results: 1) Increased alveolar-capillary permeability was observed 6 hours after endotoxin injection, which was normalized after 24 hours, and this increase was attenuated by pretreatment with DMTU and DFX. 2) Neutrophil sequestration into the lung was observed 24 hours after endotoxin administration, but this was not influenced by DMTU and DFX pretreatment. Conclusion: These results suggest that hydroxyl radical would not be involved in the sequestration of neutrophils into the lung, but plays an important role in the increase of alveolar-capillary permeability in the endotoxin-induced acute lung injury in rats.

• The Korean Journal of Mycology
• /
• v.29 no.1
• /
• pp.41-46
• /
• 2001

Oxidative stress and defense system of SD-rats were studied with an edible Nigerian mushroom, namely, Lentinus tuber-regium (Fries) Singer. Experimental diets prepared with Lentinus tuber-regium (LTR) instead of carbohydrates were fed to SD rats for 6 weeks. Hydrolxyl radical $({\cdot}OH)$ formations were significantly inhibited (21.7% and 16.4%, respectively). In LTR-50 and LTR-100 groups used instead of carbohydrates, and hydrogen peroxide and nitric oxide (NO) were also significantly inhibited by 10%, and $6{\sim}10%$, respectively compared with control group, but there was no significant changes in superoxide radical $({O_2}^-)$ formations in these groups. Lipid peroxide (LPO) and oxidized protein (OP) levels as an oxidative stress were desirably inhibited ($6{\sim}12%\;and\;5{\sim}13%$, respectively) in these LTR groups compared with control group. Superoxide dismutase (SOD), glutathione peroxidase (GSHPx) and catalase (CAT) activities were significantly increased ($15{\sim}50%,\;10{\sim}25%\;and\;60{\sim}90%$, respectively) in these LTR groups. These results suggest that an edible mushroom, Lentinus tuber-regium may inhibit an oxygen radicals and oxidative stresses, but may also effectively modulate an aging processes.

• The Korean Journal of Pharmacology
• /
• v.32 no.3
• /
• pp.335-345
• /
• 1996

A critical role of oxygen-derived free radicals has been implicated in ischemia/reperfusion (I/R)-induced brain damage. In this study, we have produced experimental I/R to the brains of Mongolian gerbil (Meriones unguiculatus) by a transient occlusion and release of the common carotid arteries. We have attempted to determine whether the oxidative stress is generated upon I/R and whether this oxidative stress is linked to the cell damage. Since hippocampus has been suggested as one of the most vulnerable regions of the brain to the oxidative stress, we analyzed samples from hippocampus in comparison with those from cortex. In addition, we have examined the expression of heat shock protein 70kD species (HSP70) in these regions in order to evaluate a possible role of this protein in I/R-induced brain damage. To determine whether the oxidative stress is produced upon I/R, we measured the glutathione oxidation, GSSG/ (GSH + 2xGSSG), as an index of oxidative stress. We found an increase of the glutathione oxidation primarily in hippocampus upon I/R. To determine whether this oxidative stress is linked to the cell damage, we measured the degree of lipid peroxidation upon I/R. We found an increase of lipid peroxidation in both regions. However, the magnitude of increases was greater in hippocampus than in cortex. In addition, we found that changes in both the magnitude and the temporal patterns of glutathione oxidation closely correlated with those of lipid peroxidation. Our study provides biochemical evidences that the oxidative stress is generated upon I/R and this oxidative stress is linked to the oxidative cell damage. Our study also provides evidences that the degree of oxidative stress as well as oxidative cell damage is greater in hippocampus than in cortex. We could not find difference in the basal level of HSP70 expression between hippocampus and cortex, indicating that the intrinsic vulnerability of hippocampus cannot be explained by the lower level of HSP70 expression. We did find, however, that the induction of HSP70 expression upon I/R was impaired in the hippocampus. This impairment appeared to be at the transcriptional level. These results suggest that the measurement of HSP70 induction may be employed as a useful predictor of differential cellular susceptibilities to the I/R-induced brain damage.

• Clinical and Experimental Pediatrics
• /
• v.52 no.2
• /
• pp.213-219
• /
• 2009

Purpose:Iron is a critical nutritional element that is essential for a variety of important biological processes, including cell growth and differentiation, electron transfer reactions, and oxygen transport, activation, and detoxification. Iron is also required for neoplastic cell growth due to its catalytic effects on the formation of hydroxyl radicals, suppression of host defense cell activities, and promotion of cancer cell multiplication. Chronic transfusion-dependent patients receiving chemotherapy may have iron overload, which requires iron-chelating therapy. We performed this study to demonstrate whether the iron chelating agent deferoxamine induces apoptosis in Saos-2 osteosarcoma cells, and to investigate the underlying apoptotic mechanism. Methods:To analyze the apoptotic effects of an iron chelator, cultured Saos-2 cells were treated with deferoxamine. We analyzed cell survival by trypan blue and crystal violet analysis, apoptosis by nuclear condensation, DNA fragmentation, and cell cycle analysis, and the expression of apoptotic related proteins by Western immunoblot analysis. Results:Deferoxamine inhibited the growth of Saos-2 cell in a time- and dose-dependent manner. The major mechanism for growth inhibition with the deferoxamine treatment was by the induction of apoptosis, which was supported by nuclear staining, DNA fragmentation analysis, and flow cytometric analysis. Furthermore, bcl-2 expression decreased, while bax, caspase-3, caspase-9, and PARP expression increased in Saos-2 cells treated with deferoxamine. Conclusion:These results demonstrated that the iron chelating agent deferoxamine induced growth inhibition and mitochondrial-dependent apoptosis in osteosarcoma Saos-2 cells, suggesting that iron chelating agents used in controlling neoplastic cell fate can be potentially developed as an adjuvant agent enhancing the anti-tumor effect for the treatment of osteosarcoma.

• The Korean Journal of Pharmacology
• /
• v.28 no.2
• /
• pp.201-212
• /
• 1992

Effect of exogenous taurine on HOCl, $NH_2Cl$ and other oxidants-induced degradation of hyaluronic acid was investigated. The scavenging action of taurine on HOCl, $NH_2Cl$ and other oxidants was examined. The antioxidant action of taurine was also compared with that of thiol compounds. Viscosity of hyaluronic acid was markedly decreased by HOCl and $NH_2Cl$ on a dose dependent fashion. The degradative effect of HOCl on hyaluronic acid was greater than that of $NH_2Cl$. Taurine effectively inhibited HOCl-and $NH_2Cl-induced$ degradation of hyaluronic acid in a dose dependent fashion. The degradative effect of HOCl was markedly inhibited by DMSO. $Fe^{2+}$ plus $H_2O_2-induced$ degradation of hyaluronic acid was inhibited by catalase and DMSO but not affected by taurine. The desradative action of xanthine and xanthine oxidase was effectively inhibited by SOD and catalase but not affected by taurine. HOCl was significantly decomposed by taurine, DMSO, GSH and MPG. Both absorbance of HOCl at 250 nm and absorbance of $NH_2Cl$ at 242 nm were significantly increased by the addition of taurine. Interaction of $NH_2Cl$ with GSH or MPG showed an initial peak absorbance, but these absorbances were gradually decreased with time. OH production in the presence of $Fe^{2+}$ and $H_2O_2$ was inhibited by catalase and DMSO but not affected by taurine. Taurine did not affect $^1O_2$ production by U.V. irradiation which is responsible for DABCO and DABA. GSH and MPG markedly inhibited the degradative action of HOCl. These results suggest that the protective action of taurine on oxidants-induced damages of tissue components, including degradation of hyaluronic acid may be attributable to both its scavenging action on HOCl and $NH_2Cl$ and the complex formation of taurine with HOCl or $NH_2Cl$ without scavenging action on oxygen free radicals. Sulfhydryl group of taurine appears to show partially a protective action on HOCl-and $NH_2Cl-induced$ degradation.

• Tuberculosis and Respiratory Diseases
• /
• v.43 no.1
• /
• pp.46-53
• /
• 1996

Background: Endotoxin or lipopolysaccharide(LPS) can prime phagocytic cells such as polymorphonuclear leukocytes, monocytes or animal peritoneal macrophages to generate increased amounts of secretory products such as oxygen free radicals and tumor necrosis factor, which play an important role in developing adult respiratory distress syndrome in gram negative sepsis. Human alveolar macrophages(HAM) are continuously exposed to various stimuli inhaled into the alveoli, and the response to LPS might be different in HAM. Therefore, we investigated the effect of LPS pre-exposure on HAM adhered to plastic surface and A549 cell(type II human alveolar epithelial cell line) monolayer. Methods: HAM were isolated from bronchoalveolar lavage fluid from normal lung of the patients with localized lung cancer and esophageal cancer. LPS was exposed to HAM for 2hrs before or after adherence to plastic surface of 24-well Linbro plate and A549 cell monolayer. And then HAM was stimulated with PMA(phorbol myristate acetate) or fMLP(N-formyl-methionylleucyl-phenylalanine). The amount of hydrogen peroxide($H_2O_2$) production in the supernatant was measured on the principle of peroxidase-dependent oxidation of phenol red by hydrogen peroxide. Results: LPS pre-exposure could not enhance $H_2O_2$ production in neither HAM adhered to plastic surface nor one to A549 cell monolayer. But LPS even in the absence of PMA or fMLP stimulation directly increased $H_2O_2$ release in HAM if added after the adherence to A549 cell monolayer. Conclusion: Endotoxin does not prime HAM, but may directly activate HAM adhered to alveolar epithelial cells. Further investagation will be necessary.

• Korean Journal of Veterinary Research
• /
• v.39 no.5
• /
• pp.878-895
• /
• 1999

Ischemic preconditioing (IPC), i.e., a preliminary brief episode of ischemia and reperfusion, has been shown to reduce the cell damage induced by long ischemia and reperfusion. Superoxide radical which is produced during reperfusion after ischemia was recognized as a factor of the ischemic injury and it is dismutated into $H_2O_2$ and $O_2$ by two types of intracellular superoxide dismutase (SOD), Cu,Zn-SOD in cytoplasm and Mn-SOD in mitochondria. Recently oxygen free radicals are suggested to induce the apoptosis, however mechanism of the reduced apoptosis by ischemic preconditioing was unknown, while many studies performed in mammalian heart indicated that ATP-sensitive $K^+$ ($K_{APT}$) channel activation related with the protective effects. The aim of present study is to investigate 1) whether IP upregulate the Cu,Zn-SOD and Mn-SOD activities, and 2) whether ischemic preconditioning decreases apoptosis via $K_{APT}$ channel activation in timely reperfused skeletal muscle after long ishemia. The experimental animals, Sprague-Dawley rats weighing 250~300g, were divided into 8 groups; 1) control group, 2) ischemic preconditioning only groups, 3) pinacidil, a $K_{APT}$ channel opener, treatment only groups, 4) glibenclamide, a $K_{APT}$ channel blocker, treatment only groups, 5) ischemia groups, 6) ischemia after IPC groups, 7) ischemia and pinacidil treatment groups, and 8) IP and ischemia after glibenclamide pretreatment groups. Animals of the control group were administered with the vehicle (DMSO) alone. Pinacidil (1mg/kg) was administered intravenously 5 minutes after initiation of ischemia, and glibenclamide (0.5mg/kg) was injected intravenously 20 minutes before IPC. In rats that were ischemic preconditioned, the left common iliac artery was occluded for 5 minutes followed by 5 minutes of reperfusion by three times using vascular clamp. Ischemia was done by occlusion of the same artery for 4 hours. The specimens of left rectus femoris muscle were obtained immediately (0 hour), 12 hours, 24 hours after drug administrations, IP or ischemia and reperfusion. The immunoreactivities of SOD and its alterations were observed by use of sheep antihuman Cu,Zn-SOD and Mn-SOD antibodies on the $10{\mu}m$ cryosections. The incidencies of apoptosis were observed by TUNEL methods with in situ apoptosis detection kit on $6{\mu}m$ paraffine section. The results obtained were as follows : 1. After IPC, immunoreactivities of Cu,Zn-SOD mainly in the small-sized fibers were increased by 24 hours, that of Mn-SOD at 0 hour and 24 hours. 2. No significant changes in immunoreactivities of SOD was observed in the pinacidil and in the glibenclamide treatment only groups, and in the ischemia only groups. 3. The immunoreactivities of the Cu,Zn-SOD were increased in the ischemia after IPC groups and the ischemia and pinacidil treatment groups. 4. The immunoreactivities of the Cu,Zn-SOD in the IPC and ischemia after glibenclamide pretreatment groups were not increased except for the 12 hours reperfusion group. But, Mn-SOD immunoreactivities were increased in the 0 hours, 12 hours and 24 hours after reperfusion. 5. In the control group, the IPC only groups, and the pinacidil treatment only groups, negative or trace apoptotic reactions were observed, but the positive apoptotic reaction occured in the glibenclamide treatment groups. 6. Moderate or many number of apoptosis were revealed in the ischemia groups, and also the IPC and ischemia after glibenclamide pretreatment group except for 12 hours and 24 hours after reperfusion. However, the incidence of apoptosis was decreased in the ischemia after IPC groups and in the ischemia and pinacidil treatment groups. 7. There is a coincidence between the increase of Cu,Zn-SOD immunoreactivities and the decrease of apoptosis in the presence of ischemia and reperfusion. These results suggest that the protective effects of ishemic preconditioing may related to the SOD activation, and the ischemic preconditioning decreases the apoptosis partially via $K_{APT}$ channel activation in timely reperfused rat skeletal muscle. It is also suggested that inhibition of apoptosis by IPC may related with the SOD activation.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.32 no.7
• /
• pp.1082-1087
• /
• 2003

This study was designed to investigate the effects of butanol (BuOH) fraction of pine (Pinus densiflora Sieb et Zucc) needle extract on membrane fluidity (MF), basal and induced oxygen radicals (BOR and IOR), lipid peroxide (LPO) and oxidized protein (OP) as an oxidative stress, and lipofuscin (LF) in liver membranes of Sprague-Dawley male rats. The rats were fed basic diets (control group) and experimental diets (BuOH-25, BuOH-50 and BuOH-100) prepared with 25, 50 and 100 mg added to basic diet for 45 days. MFs were significantly increased (about 16∼22%) in mitochondria of BuOH-50 and BuOH-100 groups compared with control group (p<0.01∼0.001) BOR and IOR formations in mitochondria were significantly decreased (11∼17% and 11∼28%, respectively) in these three BuOH groups (p<0.05∼0.001), while BOR and IOR formations in microsomes were significantly decreased (11∼24%) in BuOH-50 and BuOH-100 groups, and (15∼24%) in these three BuOH groups compared with control group (p<0.05∼0.001; p<0.01-0.001). LPO levels were significantly decreased (9% and 9∼13%, respectively) in mitochondria of BuOH-100 and microsomes of BuOH-50 and BuOH-100 groups (p<0.05∼0.01), whereas OP levels were significantly decreased (10∼12%) in mitochondria of BuOH-50 and BuOH-100 groups compared with control group (p<0.05). LF formations were significantly decreased (8∼9%) in BuOH-50 and BuOH-100 groups (p<0.05). These results suggest that butanol fraction of pine needle extract may playa effective role in an attenuating an oxidative stress and increasing a membrane fluidity.