• Applied Microscopy
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• v.27 no.3
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• pp.333-346
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• 1997

The ischemia and reperfusion injury of the skeletal muscles is caused by generation of reactive oxygen during ischemia and reperfusion. It is well known that over 4 hours of ischemia injures the skeletal muscles irreversibly. The author has demonstrated the effects of SOD (superoxide dismutase), DMTU (dimethyl thiourea) and ischemic preconditioning on ultrastructural changes of the muscle fibers in the rectus femoris muscles after 4 hours of ischemia and 1 day and 3 days of reperfusion. A total of 72 healthy Sprague-Dawley rats weighing from 200 gm to 250 gm were used as experimental animals. Under urethane(1.15 g/kg, IP, 2 times) anesthesia, lower abdominal incision was done and the left common iliac artery was occluded by using vascular clamp for 4 hours. The left rectus femoris muscles were obtained at 1 and 3 days after the removal of vascular clamp. The SOD (15,000 unit/kg) or DMTU (500 mg/kg) were administered intraperitoneally at 1 hour before induction of ischemia. The ischemic preconditioned group underwent three episodes of 5 minutes occlusion and 5 minutes reperfusion followed by 4 hours of ischemia and 1 day and 3 days of reperfusion. The specimens were sliced into $1mm^3$ and prepared by routine methods for electron microscopic observation. All specimens were stained with uranyl acetate and lead citrate and then observed with Hitachi-600 transmission electron microscope. The results were as follows: 1. SOD or DMTU alone did not affect the ultrastructure of muscle fibers in the rectus femoris muscles. The electron density of mitochondrial matrix was decreased by ischemic preconditioning. 2. Dilated cisternae of sarcoplasmic reticulum, triad, mitochondria and the loss of myofilament in the sarcomere were observed in the 4 hours ischemia and 1 day reperfused rectus femoris muscles. Markedly changed sarcoplasmic reticulum, triad, disordered or loss of myofilament, indistinct A-band and I-band, and irregular electron lucent M -line and Z-line are seen in the 4 hours ischemia and 3 days reperfused rectus femoris muscles. 3. SOD reduced the changes of organelles in the muscle fibers of the 4 hours ischemia and 1 day reperfused rectus femoris muscles of the rats, but SOD did not affect the changes of muscle fibers in the 4 hours ischemia and 3 days reperfused muscles. On the other hand, DMTU markedly attenuated considerably the ultrastructural change of the 4 hours ischemia and 1 day or 3 days reperfused rectus femoris muscles. 4. By the ischemic preconditioning, the change was attenuated remarkably in the 4 hours ischemia and 1 day reperfused rectus femoris muscles. As the ischemic reperfused changes of muscle fibers were regenerated or recovered by ischemic preconditioning, the ultrastructures of them were similar to those of normal control in the 4 hours ischemia and 3 days reperfused rectus formoris muscles. Consequently, it is suggested that DMTU is stronger inhibitor to ischemic reperfused change than SOD. The ischemia and reperfusion-induced muscular damage is remarkably inhibited by ischemic preconditioning.

• Journal of Life Science
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• v.14 no.2
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• pp.280-285
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• 2004

The ovarian hormone-deficiency induced ovariectomy rat is widely used as an aging model due to its practicality, convenience, and cost effectiveness. The surgically ovariectomized rat induces reactive oxygen species (ROS) generation like aging phenomena. Free oxygen radicals have been proposed as important causative agents of aging. The purpose of this study was to investigate the effect of chondroitin sulfate (CS) to prevent ovariectomy (OVX)-induced oxidative stress. The OVX rats were given intraperitoneally CS at doses of 100 mg/kg and 200 mg/kg daily for fifteen weeks. Malondialdehyde (MDA) levels were determined as well as the activities of superoxide dismutase (SOD), catalase (CAT), reduced-glutathione (GSH), oxidized-glutathione (GSSG), glutathione peroxidase (GPx) in the liver. The liver antioxidative enzyme activity was elevated while MDA concentration decreased in all CS treated animals. The results demonstrated that CS reduced oxidative stress in a dose dependent manner. These results suggest that CS might be a useful candidate for antioxidative reagent.

• Journal of Life Science
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• v.28 no.10
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• pp.1193-1200
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• 2018

This study investigated the physiochemical properties, antioxidative, nitrite-scavenging, and alcohol metabolism enzyme activities of nectarine vinegar prepared by a traditional fermentation method. The pH of nectarine vinegar was 3.70, the sugar content was $8.87^{\circ}Brix$, and the total acidity was 6.29%. Among organic acids detected, acetic acid was highest at 32.42 mg/ml, followed by lactic acid, malic acid, and succinic acid. Total phenol content of the nectarine vinegar was $121.84{\mu}g$ tannic acid equivalents (TAE)/100 ml. The antioxidative effects of muskmelon vinegar were measured using 1,1-Diphenyl2-picrylhydrazy (DPPH) radical-scavenging activity and superoxide dismutase (SOD) assay. DPPH of nectarine vinegar was increased in a dose-dependent manner, which was 84.47% at 40% concentration. SOD activity was increased in a dose-dependent manner, which was 89.06% at 60% concentration. Nitric scavenging activities of nectarine vinegar were 94.17%, 76.91%, and 20.21% at pH values 1.2, 3.0, and 6.0 at 100% concentration, respectively. The effects of nectarine vinegar on alcohol-metabolism were determined by measuring the generation of reduced nicotinamide adenine dinucleotide (NADH) by alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH). ADH and ALDH activities of nectarine vinegar were increased in a dose-dependent manner, which were 153.61% and 178.20 % at 60% concentration, respectively. These results suggest that nectarine vinegar has great potential as a resource for high quality functional health beverages.

• The Journal of Korean Medicine
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• v.28 no.1 s.69
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• pp.237-248
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• 2007

Objectives : Diabetes is a disease in which the body does not produce or properly use insulin. Etiological studies of diabetes and its complications showed that oxidative stress might play a major role. Therefore, many efforts have been tried to regulate free oxygen radicals for treating diabetes and its complications. Iksujisundan has been known to be effective for the treatment of diabetes. The present study was carried out to investigate the effect of Iksujisundan on renal function, peroxynitrite(ONOO-) scavenging activity and polyol pathway in streptozotocin-induced diabetic rats. Methods : The crushed Iksujisundan was extracted 3 times, each time with 3 volumes of methyl alcohol at 60$^{\circ}C$ for 24 h. The extract was filtered and evaporated under a reduced pressure using a rotary evaporator to yield 87.8g. Iksujisundan extract was orally administreted at 100 mg per 1 kg of body weight for 20 days to the diabetic rats induced by streptozotocin(60mg/kg). The effects of Iksujisundan extract on the streptozotocin-induced diabetic rats were observed by measuring the serum level of glucose, insulin, lipid components, creatinine and BUN, and also the kidney levels of superoxide anion radical(${\cdot}$O2-), nitric oxide(NO) and ONOO-, and also the enzyme activities involved in the polyol pathway. Results : The effects of Iksujisundan on the streptozotocin-induced diabetic rats with regards to body weight, blood glucose and indulin levels, creatinine and BUN levels, total cholesterol and triglyceride lavels, and HDL-cholesterol levels were all shown to be good enough to prevent and cure the diabetes and its complications. Iksujisundan inhibited the generation of ${\cdot}$O2-,NO and ONOO- in the kidney of streptozotocin-induced diabetic rats. Renal aldose reductase and sorbitol dehydrogenase activities were increased in the streptozotocin-induced diabetic rats were reversed toward natural activities. Conclusions : Iksujisundan might inhibit the development of diabetes and its complications by scavenging reactive oxygen and nitrogen species, thereby by reducing oxidative stresses and also by regulating the activities of polyol pathway enzymes, all of which could help to recover kidney function.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.2
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• pp.154-159
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• 2009

The hepatoprotective effect of ethanol extract from Hovenia dulcis fruit (HD) against ethanol-induced oxidative damage was investigated. Ethanol-induced reactive oxygen species (ROS) generation and liver damage on HepG2/2E1 cells were protected by $100{\mu}g/mL$ ethanolic extract from HD. Male C57BL/6 mice were divided into 3 groups; control (NC), ethanol (ET), ethanol plus 1 g/kg body weight ethanolic extract of HD (ET-HD). The activities of serum alanine amintransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) were significantly increased in ethanol-treated group. However, ET-HD group showed protective effect by lowering serum activities. The ET group markedly decreased the activities of catalase (CAT), superoxide dismutase (SOD), and glutathione-s-transferase (GST) with the reduced level of glutathione (GSH) in liver. On the other hand, ET-HD group increased the activities of SOD and GST, and the level of GSH. Lipid peroxidation level, which was increased after ethanol administration, was significantly reduced in ET-HD group. Based upon these results, it could be assumed that ethanolic extract of HD protected the liver against ethanol-induced oxidative damage by possibly inhibiting the suppression of antioxidant activity and reducing the rate of lipid peroxidation in vitro and in vivo. Therefore, extract of Hovenia dulcis fruit might be used as a protective agent for ethanol-induced hepatic damages.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.31 no.2 s.51
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• pp.197-206
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• 2005

In the present study, the Chuncheon buckwheat extracts prepared from its seed coats, seeds and stems were used to determine anti-oxidative effects, the content of rutin and phytic acid, and the protective role against cadmium at the cellular level. futhermore, it was evaluated whether the buckwheat, mainly known as a healthy food source, might be applicable to functional cosmetics. Up to $100 {\mu}g/mL$ of the extract was not toxic in HaCaT and B16F10 cell lines using MTT assay. The anti-oxidative capacity of superoxide radicals was shown in seed coats extracts > stem extracts=seed extracts. Although its content of rutin, known as one of effective anti-oxidants, mainly exists in the stem, any extract did not eliminate hydroxyl radicals. Phytic acid, known as a heavy metal-chelate agent, was highly concentrated in the stem. The Chuncheon buckwheat extract had $10\%$ protective effect against the treatment of $50{\mu}M$ cadmium at which $50\%$ of HaCaT cells survived. Confocal laser scanning microscope revealed the intracellular generation of reactive oxygen species (ROS) by cadmium treatment. Finally, we identified that the stem extract had the most protective effect on the elimination of ROS.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.3
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• pp.671-678
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• 2007

Diabetes is a disease in which the body does not produce or properly use insulin. Etiological studies of diabetes and its complications showed that oxidative stress might play a major role. Therefore, many efforts have been tried to regulate free oxygen radicals for treating diabetes and its complications. Gamigukihwandong-hwan has been known to be effective for the treatment of diabetes. The present study was carried out to investigate the effect of Gamigukihwandong-hwan on renal function, peroxynitrite (ONOO$^-$) scavenging activity and polyol pathway in streptozotocin-induced diabetic rats. The crushed Gamigukihwandong-hwan was extracted 3 times, each time with 3 volumes of methyl alcohol at 60$^{\circ}C$ for 24 h. The extract was filtered and evaporated under a reduced pressure using a rotary evaporator to yield 74.95 g. Gamigukihwandong-hwan extract was oral-administered 100 mg per 1 kg of body weight for 20 days to the diabetic rats induced by streptozotocin (60 mg/kg). The effects of Gamigukihwandong-hwan extract on the streptozotocin-induced diabetic rats were observed by measuring the serum level of glucose, insulin, lipid components, creatinine and BUN, and also the kidney levels of superoxide anion radical (${\cdot}O_2^-$), nitric oxide (NO) and ONOO$^-$, and also the enzyme activities involved in polyol pathway. The Effects of Gamigukihwandong-hwan on the streptozotocin-induced diabetic rats with regards to body weight, blood glucose and insulin levels, creatinine and BUN levels, total cholesterol and triglyceride levels, and HDL-cholesterol levels were all shown to be good enough to cure and prevent the diabetes and its complications. Gamigukihwandong-hwan inhibited the generation of ${\cdot}O_2^-$, NO and ONOO$^-$ in the kidney of streptozotocin-induced diabetic rats. Renal aldose reductase and sorbitol dehydrogenase activities were increased in the streptozotocin-induced diabetic rats, whereas the ones in the Gamigukihwandong-hwan administered group among the streptozotocin-induced diabetic rats were reversed toward the natural activities. Gamigukihwandong-hwan might inhibit the development of diabetic nephropathy by scavenging reactive oxygen and nitrogen species, thereby by reducing oxidative stresses and also by regulating the activities of polyol pathway enzymes, all of which could help to recover the function of kidney.

• Journal of Life Science
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• v.32 no.4
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• pp.290-297
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• 2022

This study was performed to investigate the organic acids, alcohol metabolism enzyme, and antioxidative, nitrite-scavenging, and hepatoprotective effects of Dendropanax morbifera vinegar prepared by a traditional fermentation method. Among the organic acids detected, acetic acid was the highest found, at 91.72 mg/ml, followed by lactic acid (7.31 mg/ml), malic acid (1.36 mg/ml), and succinic acid (1.20 mg/ml). The total polyphenol content of the D. morbifera vinegar was 13.73 ㎍ tannic acid equivalent (TAE)/ml. The 1,1-Diphenyl-2-picrylhydrazy (DPPH) radical scavenging activity of D. morbifera vinegar was 76.04% at a 60% concentration. The superoxide dismutase (SOD) activity of D. morbifera vinegar was increased in a dose-dependent manner, which was 95.14% at a 60% concentration, while the α-glucosidase inhibitory activity of D. morbifera vinegar was 98.94% at a 10% concentration. The effects of D. morbifera vinegar on alcohol metabolism were determined by measuring the generation of reduced nicotinamide adenine dinucleotide (NADH) by alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH). The ADH and ALDH activities of D. morbifera vinegar were increased in a dose-dependent manner, 43.62% and 60.39% at a 60% concentration, respectively. The D. morbifera vinegar showed significant protective effects against tacrine-induced cytotoxicity in HepG2 cells at the 0.6% concentration. These results suggest that D. morbifera vinegar has great potential as a resource for high quality functional health beverages.

• Journal of Korean Medicine Rehabilitation
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• v.34 no.2
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• pp.15-28
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• 2024

Objectives We used the D-galactose (D-gal) induced C2C12 myoblast senescence model to investigate whether ethanol extract of Perilla. fructescens leaves (EEPF) could delay cellular senescence and regulate related mechanisms. Methods C2C12 myogenic cells were cultured in an incubator under 37 ℃ and 5% CO₂ conditions. EEPF, dried perilla leaves were pulverized and extracted at 1:10 (v/v) at 50 ℃ for 4 hours. Cell counting kit-8 and western blot analysis was performed. Annexin V-FITC apoptosis detection kit and DAPI staining was applied. Catalase (CAT), glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC), superoxide dismutase (SOD), and malondialdehyde analysis kits were used. To measure the level of reactive oxygen species generation, staining and flow cytometry was used. To analyze the mitochondrial activity, membrane potential changes were measured using JC-1. 𝛽-gal activity was analyzed using SA-𝛽-gal staining solution, and DNA damage was analyzed by using 𝛾-H2AX. Quantikine ELISA kit was used to analyze inflammatory cytokine production. Results According to the results of this study, EEPF significantly alleviated the decrease in cell viability in C2C12 cells treated with D-gal and suppressed the decrease in the expression of proliferating cell nuclear antigen. EEPF also markedly blocked D-gal-induced C2C12 cell apoptosis and restored reduced activity of CAT, GSH-Px, T-AOC, SOD. In addition, EEPF suppressed the decrease in 𝛽-galactosidase activity, the induction of DNA damage and the increase in expression of senescence-associated secretory phenotype proteins such as p16, p53 and p21 in D-gal-treated C2C12 cells. Furthermore, EEPF significantly attenuated D-gal-induced production and expression of inflammatory cytokines such as interleukin (IL)-6 and IL-18. Conclusions The results of this study indicate that EEPF can be used as a potential candidate for the prevention and treatment of muscle aging.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.8
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• pp.979-984
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• 2006

Although iron is essential for many physiological processes, excess iron can lead to tissue damage by promoting the generation of reactive oxygen species (ROS). There is increasing evidence that ROS might play an important role in the pathogenesis of cardiovascular disease. However, the effects of iron excess on platelet function and the thrombotic response to vascular injury are not well understood. We examined the effects of iron excess-induced oxidative stress and the antioxidants on platelet aggregation. Oxidative stress was accessed by either free iron $(Fe^{+2})$ or hydrogen peroxide $(H_2O_2)$, as well as their combination on washed rabbit platelets (WPs) in vitro. When WPs were stimulated with either $Fe^{+2}$ alone or a subthreshold concentration of collagen, which gave an aggregatory curve with a little effect, and a dose dependent increase in platelet aggregation was observed by increasing concentrations of $Fe^{+2}$ with $H_2O_2$. This aggregation was associated with the iron-catalyzed formation of hydroxyl radicals from $H_2O_2$, and were inhibited by NAD/NADP (proton acceptor), catalase $(H_2O_2\;scavenger)$, tiron (iron chelator), mannitol (hydroxyl radical scavenger), and indomethacin (cyclooxygenase inhibitor), but not by NADH/NADPH (proton donor), superoxide mutase, and aspirin. However, NADH/NADPH, an essential cofactor for the antioxidant capacity by the supply of reducing potentials, showed the effect of an enhanced radical formation, suggesting a role for NADH/NADPH-dependent oxidase. These results suggest that iron $(Fe^{+2})$ can directly interact with washed rabbit platelets and this aggregation be mediated by OH formation as in the Fenton reaction, inhibited by radical scavengers.