• Journal of Embryo Transfer
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• v.22 no.3
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• pp.155-160
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• 2007

During in vitro culture of mammalian oocytes and embryos, the cells are exposed to the risks that cause cell injury or death. Numerous studies have been reported that the cell injury may be induced by the action of free radicals generated by auto-oxidation. This study was undertaken to investigate the optimal culture condition system for in vitro culture of porcine embryos. We first evaluated the effect of culture media on the porcine embryo development. NCSU-23 and PZM-5, culture medium tested, were failed to produce significant difference on the rate of blastocyst formation. In NCSU-23, the developmental rate was slightly higher than that in PZM-5. During in vitro maturation (IVM), fertilizaton (IVF), and culture (IVC) under 5 or 20% oxygen ($O_2$), the rates of cleavage and development were insignificantly different from each other under our culture condition (20% $O_2$, in NCSU-23), the mean cell number per blastocyst was $40{\pm}10$. These results showed that medium and $O_2$ concentration had no significant effect on the development of porcine embryos.

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

• Korean Journal of Plant Resources
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• v.22 no.4
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• pp.349-358
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• 2009

The potential use of 4 plant species, Astragalus membranaceus var. membranaceus, Senna occidentalis, Dianthus longicalyx and Plantago asiatica, as new sprout vegetables with high antioxidant function was examined in the present experiments. Seeds of above plants were allowed to germinate under light condition, and seedlings were maintained under dark condition for shoot growth in length for contain period of time. Then the seedlings were put under light for photosynthesis (greening treatment) for the period of 0${\sim}$3 days. Samples were collected to analyze the changes in antioxidant levels and activity, and it was observed that antioxidant substances were affected by greening treatments, depending on plant species. In A. membranaceus, the contents of total polyphenol was highest with no greening, total flavonoids with 3 days greening, DPPH radical scavenging effects with no greening, ABTS scavenging with 1 day greening, $Fe^{2+}$ chelating effects with no greening, and inhibitory activity against linoleic acid peroxidation with 3 day greening. In S. occidentalis, highest levels of antioxidant activity and radical scavenging effects were obtained by 2 day greening, $Fe^{2+}$ chelating effects by no greening and inhibitory activity against linoleic acid peroxidation by 1 day greening. In D. longicalyx, highest levels of antioxidant activity and $Fe^{2+}$ chelating effects were obtained by 2 day greening, $Fe^{2+}$ chelating effects by no greening and inhibitory activity against linoleic acid peroxidation by 1 day greening. In D. longicalyx, highest levels antioxidant activity and $Fe^{2+}$ chelating effects were observed with 3 day greening, and highest radical scavenging effects and inhibitory activity against linoleic acid peroxidation with no greening treatment. In P. asiatica, antioxidant activity and radicals scavenging effects were highest with 2 day greening, whereas highest chelating effects was obtained with no greening and highset inhibitory activity against linoleic acid peroxidation with 3 day greening. As the length of greening treatments influenced the antioxidant levels and function in plant species tested in this experiments, different culture methods are recommended for different plant species to get maximum health benefits out of sprout vegetables.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.5
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• pp.739-744
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• 2003

This study was carried out to investigate the antioxidative activity and to identify the active components of hot-water extract of Paeoniajaponica (PJ), which was a main ingredient of a herb mixture preparation recently established as a potent candidate of radioprotector in our laboratory. The water extract was fractionated with CHCl$_3$, EtOAc and n-BuOH. The extract and its fractions showed very low activity in hydroxyl radical scavenging test. In lipid peroxidation test, the extract, EtOAc and water fractions showed moderate inhibition with the ratio above 50%. In DPPH radical scavenging test, the extract, EtOAc and water fraction showed high activity with the ratio above 80%, especially. EtOAc fraction scavenged the radicals as much as synthetic antioxidant (BHA), even at low concentration. It is suggested that mai or partition for antioxidative activity of Paeonia japonica was EtOAc fraction. Subsequently, two active compounds (PJE021-1 and JE024-1) from EtOAc fraction were isolated by using MCI gel and silica gel column chromatography The two compounds inhibited remarkedly the $H_2O$$_2$-induced DNA damage in human peripheral blood lymphocytes, measured by single-cell gel electrophoresis (SCGE). PJE021-1 protected the cells to almost negative control level, dose-dependently. PJE024-1 exhibited a potent inhibition with the ratio of 71% at even low concentration (0.5 $\mu\textrm{g}$/$m\ell$). Finally, their chemical structures were identified as gallic acid (PJE021-1) and (＋)-catechin (PJE024-1), respectively, on the basis of the speculation of spectral and physical data.

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

• Journal of Yeungnam Medical Science
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• v.11 no.2
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• pp.262-269
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• 1994

Glutathione (GSH) is a well-known antioxidative cellular component which is ubiquitous in nature. Several enzymes involved in GSH metabolism and recycling have been found to play important roles in detoxification of xenobiotics and free radicals. In this study, total GSH content, activity of GSH peroxidase and GSH reductase were measured in liver and kidney of cisplatin treated rats. Total GSH content (mM/g protein) of liver was higher in cisplatin treated rats ($1.51{\pm}0.28$) than of nontreated control ($0.95{\pm}0.28$), and in kidney, it was also higher in cisplatin treated rats ($0.87{\pm}0.20$) than that of control ($0.68{\pm}0.14$). The activity of GSH peroxidase (${\mu}M/mg$ protein/min) was lower in liver of cisplatin treated rats ($348.0{\pm}18.54$) than that of control ($415.5{\pm}53.15$), in kidney it was increase din cisplatin treated rats ($380.5{\pm}51.86$) compared to control ($327.3{\pm}20.36$). The activity of GSH reductase (${\mu}M/mg$ protein/min) was higher in liver of cisplatin treated rats ($3.09{\pm}0.88$) than that of control ($2.28{\pm}0.61$), in kidney it was also higher in cisplatin treated rats ($8.50{\pm}2.62$) than that of control ($3.30{\pm}1.10$). In summary, detoxification of ciplatin was revealed lesser effect in kidney as show increasion of GSH peroxidase and reductase and detoxification of cisplatin was expressed effectively in liver by increasing of GSH content and decreasing GSH peroxidase.

• Clinical and Experimental Pediatrics
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• v.52 no.2
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• pp.213-219
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• 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
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• v.24 no.1
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• pp.53-61
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• 1988

The effect of antioxidants on the myocardial cellular damage which occurs during reoxygenation of hypoxic myocardium was examined in isolated rat hearts. The roles of oxygen free radical and lipid peroxidation in reoxygenation injury of myocardium were also investigated. In Langenorff preparation of isolated rat heart, which was made hypoxic by perfusion with the substrate free, hypoxic cardioplegic solution ($37^{\circ}C$, 90 min), the release of cytosolic enzymes (creatine phosphokinase, lactic dehydrogenase) and a lipid peroxidation product, malondialdehyde into the coronary effluent were abruptly increased by reoxygenation. The release of enzymes was closely parallel to that of MDA. These increases of enzymes and lipid peroxidation product were suppressed to various degrees in the presence of scavengers of superoxide anion (superoxide dismutase, 10,000 U), hydrogen peroxide (catalase, 25,000 U) and hydroxyl radical (dimethyl sulfoxide, 10%). A natural antioxidant, ${\alpha}-tocopherol$(4.5 uM) and a synthetic one, butylated hydroxytoluene (2 uM) suppressed the release of cytosolic enzymes with the concomittent reduction of lipid peroxidation as measured by malondialdehyde release into the coronary effluent. These effects of antioxidants were dose dependent, and were more pronounced when the antioxidants were administered throughout hypoxic and reoxygenation periods than given during reoxygenation period only. These results suggest that cytotoxic oxygen free radicals produced in the myocardium during reoxygenation may be responsible fur the myocardial cellular injury by enhancing the lipid peroxidation of cellular membranes. Furthermore, the antioxidants may exert protective effect against reoxygenation damage of hypoxic myocardium through the inhibition of lipid peroxidation reaction.

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

• The Korean Journal of Pharmacology
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• v.30 no.3
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• pp.321-330
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• 1994

The protective effect of 'ischemic preconditioning (PC)' on ischemia-reperfusion injury of heart has been reported in various animal species, but without known mechanisms in detail. In an attempt to investigate the cardioprotective mechanism of PC, we examined the effects of PC on the myocardial oxidative injuries and the oxygen free radical production in the ischemia-reperfusion model of isolated Langendorff preparations of rat hearts. PC was performed with three episodes of 5 min ischemia and 5 min reperfusion before the induction of prolonged ischemia (30 min)-reperfusion(20 min). PC prevented the depression of cardiac function (left ventricular pressure x heart rate) observed in the ischemic-reperfused heart, and reduced the release of lactate dehydrogenase during the reperfusion period. On electron microscopic pictures, myocardial ultrastructures were relatively well preserved in PC hearts as compared with non-PC ischemic-reperfused hearts. In PC hearts, lipid peroxidation of myocardial tissue as estimated from malondialdehyde production was markedly reduced. PC did not affect the activity of xanthine oxidase which is a major source of oxygen radicals in the ischemic rat hearts, but the myocardial content of hypoxanthine (a substrate for xanthine oxidase) was much lower in PC hearts. It is suggested from these results that PC brings about significant myocardial protection in ischemic-reperfused heart and this effect may be related to the suppression of oxygen free radical reactions.