• Title/Summary/Keyword: Free radicals

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Priming Effect of Endotoxin in Human Alveolar Macrophage (사람 폐포대식세포에서 내독소의 Priming 효과)

  • Chung, Man-Pyo;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.1
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    • pp.46-53
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    • 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.

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Immunocytochemistry of Metallothionein Expression in Developing Rat Liver (발생중인 흰쥐 간에서의 Metallothionein 발현에 관한 면역세포화학적 연구)

  • Oh, Seung-Han;Ahn, Young-Mo;Shin, Kil-Sang;Kim, Wan-Jong
    • Applied Microscopy
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    • v.34 no.3
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    • pp.171-178
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    • 2004
  • Metallothionein (MT) is a family of ubiquitous, low molecular weight (6-7 kDa), cysteine-rich protein with a high affinity to metal ions and has no aromatic amino acids and histidine. Some of the known functions of MT include detoxification of heavy metals and alkylating agents and neutralization of free radicals. Also, this protein may affect a number of cellular processes including gene expression, apoptosis, proliferation and differentiation. But, its actual functions are still not clear. The present study was undertaken to examine immunocytochemically the localization of MT in developing rat liver. On the day 11 of gestation, the fetal rat liver has already been formed and contained numerous oval cells with high nuclear cytoplasmic ratio, which were the progenitors of hepatic parenchymal cells, but no reaction products of MT were detected at this time. And then, positive reactions against MT started to appear predominantly in the parenchymal cells of liver from the 13th day after gestation. Reaction products, immunogold particles or brown coloration, were localized at both the nucleus and the cytoplasm of the parenchymal cells, except mitochondria. The intensity of this reaction gradually increased, and exhibited the strongest at birth. The intensity of MT staining and immunogold labelling diminished with growth, and by the 15th day after birth weak positive reaction was observed in the cells. In brief, positive reactions for MT were observed in the oval cells and the parenchymal cells during fetal stage, meanwhile they were present only in the parenchymal cells after birth. The present results suggest that MT possibly involves parechymal cell proliferation and differentiation through the storage or the supply of various metal ions in the developing rat liver.

Effects of ischemic preconditioning, KATP channel on the SOD activation and apoptosis in ischemic reperfused skeletal muscle of rat (허혈양상화와 KATP 통로가 허혈후 재관류된 흰쥐의 골격근육에서 SOD 활성 및 apoptosis에 미치는 영향)

  • Abn, Dong-choon;Paik, Doo-jin;Yang, Hong-hyun
    • Korean Journal of Veterinary Research
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    • v.39 no.5
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    • pp.878-895
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    • 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.

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Detection Characteristics of Gamma-Irradiated Korean Medicinal Herbs by Using PSL, TL, and ESR (PSL, TL 및 ESR 분석에 의한 감마선 조사 한약재의 검지 특성)

  • Yang, Hee-Sun;Park, Yong-Dae;Jin, Chang-Hyun;Choi, Dae-Seong;Chung, Hyung-Wook;Byun, Myung-Woo;Jeong, Il-Yun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.11
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    • pp.1529-1533
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    • 2008
  • The detection characteristics of gamma-irradiated ($0{\sim}10.0\;kGy$) medicinal herbs (Platycodon grandiflorum, Acanthopanax chiisanensis) were investigated by photostimulated luminescence (PSL), thermoluminescence (TL), and electron spin resonance (ESR). The results of the PSL, a first screening method in comparison with the TL, showed photon counts greater than 5,000 counts/60 s (positive) in the irradiated samples, while the non-irradiated samples yielded photon counts less than 700 counts/60 s (negative). The TL was also applied for the detection method of irradiated medicinal herbs and showed that the non-irradiated sample revealed a glow curve with a low intensity, while the irradiated samples showed a higher intensity. These results were normalized by re-irradiating the mineral grains with a irradiation dose of 1.0 kGy, and a second glow curve was recorded. The ratio of the intensity of the first glow curve ($TL_1$) to that after the normalization dose ($TL_2$) was determined and compared with the recommended threshold values. TL ratio ($TL_1/TL_2$) was below 0.007 for the non-irradiated sample and higher than 0.1 for all irradiated samples (above 1.0 kGy). ESR spectroscopy revealed specific signals (6.065 mT) derived from free radicals in cellulose containing irradiated medicinal herbs. The P. grandiflorum showed clearer signals than A. chiisanensis. From the results of our studies, the PSL, TL, and ESR determinations were found to be suitable for the detection of irradiated medicinal herbs such as P. grandiflorum and A. chiisanensis.

Anti-wrinkle Effect of Cambodian Phellinus linteus Extracts (캄보디안 상황버섯 추출물의 주름개선 효과 연구)

  • Cheon, Soon-Ju;Jang, Min-Jung;Jang, Young-Ah;Choi, Eun-Young;Jun, Dong-Ha;Kim, Young-Hun;Cho, Woo-A;Jeong, Yeon-Sook;Kwon, Hyeork-Bum;Kim, Tae-Hoon;Choi, Kyung-Im;Do, Jeong-Ryong;Lee, Chang-Eon;Lee, Jin-Tae
    • Journal of Life Science
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    • v.18 no.12
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    • pp.1718-1721
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    • 2008
  • The skin of human is constantly being exposed to environmental irritants such as ultraviolet, smoke and chemicals. These irritants cause free radicals and reactive oxygen species which leave serious damages on the cells of skin. The water and ethanol extracts of Cambodian Phellinus linteus were investigated for the activities of anti-lipid peroxidation and anti-wrinkle effects to apply as a functional ingredient for cosmetic products. As the result of evaluation of liquid oxidation rate by add $Fe^{2+}$ and $Cu^{2+}$ to Cambodian Phellinus linteus extracts, Cambodian Phellinus linteus ethanol extracts were higher than Cambodian Phellinus linteus water extracts in the chealting ability of $Fe^{2+}$ and $Cu^{2+}$. The Cambodian Phellinus linteus ethanol extracts exhibited that anti-lipid peroxidation higher than butylated hydroxytoluene (BHT) at the concentration of 0.1 mg/ml, 0.5 mg/ml and 1 mg/ml. Cambodian Phellinus linteus water and ethanol extracts showed a higher inhibitory effect on $Fe^{2+}$-induced lipid peroxidation compared to $Cu^{2+}$-induced lipid peroxidation. In the case of anti-wrinkle effect, the elastase inhibition activity of Cambodian Phellinus linteus ethanol extracts was 50.7%, and it is higher than urosolic acid at the concentration of 0.01 mg/ml. Also, in collagenase inhibition activity, Cambodian Phellinus linteus water extract showed low effect, but Cambodian Phellinus linteus ethanol extract was about 50% at a 0.1 mg/ml. concentration. These results proved that Cambodian Phellinus linteus had anti-lipid peroxidation and anti-wrinkle effect. Therefore, Cambodian Phellinus linteus could be useful as an anti-wrinkle cosmetic ingredient.

Iron Status According to Serum Selenium Concentration and Physique in Young Female Adults (젊은 여성의 혈청 셀레늄 농도 및 체격에 따른 체내 철 수준)

  • Lee, Ok-Hee;Chung, Yong-Sam;Moon, Jong-Wha
    • Journal of Nutrition and Health
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    • v.43 no.2
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    • pp.114-122
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    • 2010
  • Se and Fe are trace minerals acting as antioxidant scavenging free radicals. Iron deficiency is the most frequently reported nutritional deficiency in females. Body iron status are known to be dependent not only upon dietary iron intake, but also upon micro-mineral nutrition and obesity. Antioxidants such as selenium are reported to play an important role on the regulation of erythropoiesis by protecting RBC membrane from antioxidative damage. In this study, iron status in young females and its relationships with selenium status and physique were examined. Serum selenium and iron concentrations were measured by HANARO research reactor using neutron activation analysis method (NAA-method). The proportion with iron deficiency and anemia were 27.1% and 8.6%, respectively in young females, but the proportion with iron deficient anemia was 1.4%. The mean serum selenium level was $12.0\;{\mu}g/dL$ and in normal range in the young women. The study participants were tertiled according to BMI and serum selenium levels. Serum ferritin and iron levels inclined with increasing BMI tertiles. Serum iron and RBC count were higher in middle selenium group than low selenium group. Individuals had significantly lower hematocrit level in the lowest tertile for their serum selenium levels compared with the highest tertile. The serum ferritin level was predicted 25% by BMI and RBC count 26.2% by the serum selenium level and body fat%. In conclusion, this study shows that body iron status in young adult females are influenced by obesity and body selenium status.

The Composition of the Root of Ixeris dentata var. albiflora Nakai. and Cell Viability and DPPH Radical Scavenging Activities of its Extract (흰씀바귀 (고채(苦菜), Ixeris dentata var. albiflora Nakai)뿌리의 성분 분석과 추출물의 세포 생존율 및 DPPH 라디칼 소거 활성)

  • Hong, Seul-Gi;Jeong, Dong-Myong;Kim, Ki-Young;Hwang, Eun-Hee
    • Journal of Nutrition and Health
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    • v.43 no.2
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    • pp.105-113
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    • 2010
  • Ixeris dentata var. albiflora Nakai, a herbal plant, is often used to make a strong stomach as an antiphlogistic used when dyspepsia and to improve appetite in Korea and China. And also it is used for adult diseases such as diabetes and liver diseases as Korean traditional medicine. In this study, the composition and DPPH radical scavenging activities of the root of Ixeris dentata var. albiflora Nakai and its effects on cell viability on vero and chang cells were investigated. Moisture, crude ash, crude protein and crude lipid were 79.14, 2.49, 8.28 and 2.56 g/100 g respectively. The highest mineral content was K. The major free sugars were glucose, fructose and sucrose. Major fatty acid are linoleic acid, palmic acid and linolenic acid. Major amino acids were glutamic acid, arginine and aspartic acid and the total contents of amino acids were 28.12 mg/g. The methanol extracts were further fractionated with n-hexane, chloroform, ethylacetate, butanol and water to get an active fraction. In addition, cell viabilities in each fraction were determined. Methanol extract, butanol, and aqueous fraction showed strong survival rates in vero cell and chang cell viability test, and hexane, chloroform, and ethylacetate fraction were examined for toxin in a cell. The root of Ixeris dentata var. albiflora Nakai had scavenging activities against DPPH radicals in a dose-dependent assay. Ethylacetate fraction's SC50 was $6.8\{\mu}g/mL$, very strong DPPH radical scavenging activities, but water fraction did not show any activity.

Effect of PLA2 Inhibitor Rutin on Endotoxin-Induced Acute Lung Injury (내독소로 유도된 급성폐손상에서 PLA2의 억제제인 Rutin의 효과)

  • Kim, Seong-Eun;Lee, Young-Man;Park, Won-Hark
    • Applied Microscopy
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    • v.34 no.1
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    • pp.31-42
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    • 2004
  • Acute respiratory distress syndrome (ARDS) is a kind of acute lung injury characterized by inflammatory disruption of alveolar-capillary barrier and notorious for its high mortality. Neutrophils cause cell damage through the production of free radicals, inflammatory mediators, and proteases in ARDS. $PLA_2$ might serve a primary regulatory role in the activation of neutrophils. This present study was performed to elucidate the effect of rutin known as $PLA_2$ inhibitor on ARDS induced by endotoxin. Endotoxin had increased lung myeloperoxidase (MPO) activity, BAL (bronchoalveolar lavage) protein content, numbers of neutrophils in BALF (bronchoalveolar lavage fluid) compared with those of control rat (p<0.001). In addition, histological evidence of lung injury was correlated with neutrophil influx into alveolar space and cerrous perhydroxide granules were found in lining of endothelial cell, alveolar type I, II cells. In contrast, pretreated group of rutin had significantly decreased all of the parameters (p<0.001). These data suggest that inhibition of $PLA_2$ is one step approach that block the process of ARDS. Accordingly, we conclude that rutin can be used as the prophylactic agent for ARDS on the bases of these experimental results.

Evaluation of free radical scavenging and pancreatic lipase inhibitory effects of Aquilaria agallocha extracts (침향 추출물의 라디칼 및 췌장 지방분해 효소저해 활성 평가)

  • Lee, Ha Yeong;Lee, In-Chul;Kwak, Jae Hoon;Kim, Tae Hoon
    • Food Science and Preservation
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    • v.22 no.3
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    • pp.437-442
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    • 2015
  • In a continuing screening of selected medicinal plants native to South Korea, the antioxidant and pancreatic lipase inhibitory activities of an aqueous methanolic extract from the heartwood of Aquilaria agallocha were investigated. Eighty percent of the methanolic extract of A. agallocha was further divided into $CH_2Cl_2$, EtOAc and n-BuOH in order to yield four solvent-soluble portions, namely $CH_2Cl_2$-soluble, EtOAc-soluble, n-BuOH-soluble and $H_2O$ residue. The antioxidant properties were evaluated by employing radical scavenging assays using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) ($ABTS^+$) radicals, while the anti-obesity efficacy of A. agallocha extracts and solvent-soluble portions were tested by porcine pancreatic lipase assay. All tested samples showed dose-dependent radical scavenging and pancreatic lipase inhibitory activities. Among the tested extracts and solvent-soluble portions, the $CH_2Cl_2$-soluble portion showed much higher radical scavenging activity and pancreatic lipase inhibitory properties when compared with other solvent-soluble portions. This result suggested that there was a significant relationship between the total phenolic content and biological efficacies, and A. agallocha extract might be considered as a new potential source of natural antioxidants and as a pancreatic lipase inhibitory source. A more systematic investigation of this biomass will be performed for further investigation of activity against antioxidative and anti-obesity effects.

Effect of Dietary Supplementation of Vitamin A and Chronic Consumption of Ethanol on Oxidative Damage and Antioxidant System in Rats (비타민 A 보충 식이 및 에탄올의 만성적 급여가 흰쥐의 체내 산화적 손상과 항산화체계에 미치는 영향)

  • 양경미
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.2
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    • pp.278-286
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    • 2003
  • Alcohol is well known agent which can damage the human tissues such as liver via stimulating lipid peroxidation. On the other hand, carotenoids in addition to vitamins A, C and I play important roles in protecting these oxidative damages as well as preventing the production of free radicals. This study was carried out to investigate the effect of dietary vitamin A on lipid peroxidation and antioxidants status in ethanol-treated rats. In the experiment, male Sprague-Dawley rats weighing 160~180 g were given a liquid diet containing 36% of total calories as ethanol for 7 weeks. The pair-fed control rats received an isocaloric amount of diet containing sucrose instead of ethanol on the following day Additionally, the liquid diet contained adequate amount of $\beta$-carotene, retinyl acetate or 13-sis-reinoic acid except vitamin A-deficient diet. The results obtained are as follows. The levels of plasma and hepatic lipid peroxide were increased after chronic ethanol feeding in rats. Retinyl acetate supplementation significantly reduced lipid peroxidation induced by ethanol feeding Glucose 6-phosphatase activity was significantly reduced in rats fed vitamin A-deficient diet with ethanol and alkaline phosphatase activity was significantly induced in rats fed 13-cis-reinoic acid diet with ethanol. Catalase and alcohol dehydrogenase activities did not show a consistent tendency in experiment groups. The hepatic antioxidant enzyme activities did not significantly changed by chronic ethanol feeding groups. The striking decrease in conversion of $\beta$-carotene to retinol was observed in rats fed a $\beta$-carotene diet with ethanol feeding The level of retinol and retinoic acid in plasma and liver was decreased after chronic ethanol administration Based on this result, these data suggest that ethanol feeding enhances oxidative stress especially in those fed a vitamin A-deficient diet, and vitamin A supplementation, especially, retinyl acetate intake can prevent enhanced lipid peroxidation and related damage to some extent.