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A Study on Anti-aning of Electroacupuncture at Joksamni($ST_{36}$) in Rats (족삼리(足三里) 전침(電鍼)이 흰쥐의 노화(老化)에 미치는 영향(影響))

  • Lee, Jun-Seong;Yim, Yun-Kyoung;Kim, Young-Il
    • Journal of Acupuncture Research
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    • v.24 no.3
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    • pp.145-161
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    • 2007
  • Objectives : The purpose of this study is to investigate the effect of electroacupuncture at Joksanmi($ST_{36}$) on Rat's aging by experimental methods. Methods : The author performed several experimental procedures to observe the effects of the EC-HAS at the arthritis. First, I measured the cell survival rate of the mice lung fibroblasts. Second, the incidence rate of arthritis and arthritis index of CIA were analyzed. Third, the levels of IL-6, $INF-{\gamma}$ , $INF-{\alpha}$ , $IL-1{\beta}$, IgG, IgM and anti-collagen II were measured in serum and the level of $INF-{\gamma}$, $INF-{\gamma}$/IL-4 ratio in the CIA mouse spleen cell culture. Fourth, histological analysis of the mice joint was performed. Fifth, the expression ratio of $CD3e^+$ cells to $CD19^+$ cells, $CD4^+$ cells to $CD8^+$ cells, $CD11a^+CD19^+$ cells, $CD4^+CD25^+$ cells, $CD3^+CD69^+$ cells, and $CD11b^+Gr-1^+$ cells were checked. Results: 1. The hematocrit was meaningfully (p<0.01, p<0.05) decreased in $ST_{36}$ group in comparison with the control and sham group. 2. The glucose, ALP were meaningfully (p<0.05, p<0.05) decreased in $ST_{36}$ group in comparison with the control group. 3. In the activation of hepatic antioxidase, the catalase was meaningfully (p<0.01) increased in $ST_{36}$ group in comparison with the control group. 4. In the liver, the NO formation was meaningfully (p<0.001) decreased in $ST_{36}$ group in comparison with the nomal, control and sham group. 5. In the activation of splenetic antioxidase, the GSH was meaningfully (p<0.01) increased in $ST_{36}$ group in comparison with the sham group, and the catalase was meaningfully (p<0.001) increased in $ST_{36}$ group in comparison with the control group. 6. In the spleen, the MDA formation was meaningfully (p<0.01, p<0.05) decreased in $ST_{36}$ group in comparison with the control and sham group. 7. In the spleen cell culture, the levels of IL-6 was meaningfully (p<0.05) decreased in comparison with the sham group, $INF-{\gamma}$( was meaningfully (p<0.05) increased in comparison with the control group. Conclusions: According to the results, it is considered that electroacupuncture at Joksamni($ST_{36}$) is effective in increasing the activities of antioxidative enzyme and inhibiting lipid peroxides, and has an immunomodulatory reaction. In conclusion, it han an significant anti-aging effect and need more experimental study.

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Pituitary Adenylate Cyclase-activating Polypeptide (PACAP) Treatment during Pre-maturation Increases the Maturation of Porcine Oocytes Derived from Small Follicles

  • Park, Kyu-Mi;So, Kyoung-Ha;Hyun, Sang-Hwan
    • Journal of Embryo Transfer
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    • v.33 no.1
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    • pp.1-11
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    • 2018
  • Cellular cyclic adenosine-3' 5'-monophosphate (cAMP) modulator is known as meiotic inhibitor and can delays spontaneous maturation in IVM experiment. Among many cAMP modulators, the role of Pituitary adenylate cyclase activating polypeptide (PACAP) on IVM isn't known. The purpose of this study is to improve the maturation of oocytes derived from follicles ${\leq}3mm$ in diameter through PACAP as meiotic inhibitor during pre-in vitro maturation (pre-IVM). First, we checked PACAP and its receptors in cumulus cells and, to establish the optimal phase and concentration of PACAP for pre-IVM, we conducted chromatin configuration assessments. As a result, the rate of GV (Germinal Vesicle) according to duration of pre-IVM was significantly decreased 12 h and 18 h after IVM (87.1 and 84.1%, respectively) compared to 0 h (99.4%). When COC was cultured for 18 h, the GV rate in the $1{\mu}M$ of PACAP treatment group (82.1%) was significantly higher than any other PACAP treatment groups (60.5, 64.1, 74.4 and 69.9 %, respectively). So, we divided into four groups as follows; MF (the conventional IVM group, obtained from follicle from 3 to 6 mm in diameter), SF (the conventional IVM group, obtained from follicle ${\leq}3mm$ in diameter), Pre-SF(-)PACAP (IVM group including 18 h pre-IVM without $1{\mu}M$ of PACAP, obtained from follicle ${\leq}3mm$ in diameter) and Pre-SF(+)PACAP (IVM group including 18 h pre-IVM with $1{\mu}M$ of PACAP, obtained from follicle ${\leq}3mm$ in diameter). To examine the effect of PACAP during pre-IVM, we investigated analysis of nuclear maturation, intracellular glutathione (GSH) and reactive oxygen species (ROS) levels. In cumulus cells, PACAP receptors, ADCYAP1R1 and VIPR1 were detected but were not detected in oocytes. After IVM, the Pre-SF(+)PACAP had the highest Metaphase II rate (91.7%) among all groups (P<0.05). The GSH levels in the MF and Pre-SF(+)PACAP were significantly higher than in the other groups (P<0.05) and ROS levels was no significant difference among all groups. In conclusion, these results indicated that even though the oocytes were derived from SF, pre-IVM application of PACAP improved meiotic and cytoplasmic maturation by regulating intracellular oxidative stress.

The Effects of Pro-inflammatory Cytokines by Cisplatin on the Death of Sensory Hair Cells. (시스플라틴에 의한 염증성 사이토카인의 청각유모세포 사멸 효과)

  • Lee, Jeong-Han;Park, Chan-Ny;Park, Rae-Kil
    • Journal of Life Science
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    • v.18 no.4
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    • pp.542-549
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    • 2008
  • Cisplatin (cis-diamminedichloroplatinum II : CDDP) is the most widely used anticancer drug against a variety of human neoplasms. However, its clinical use is limited by the onset of severe side effects, including ototoxicity and nephrotoxicity. Even though a number of evidences in cytotoxic mechanism of cisplatin have been suggested, the role of pro-inflammatory cytokines in cisplatin cytotoxicity of auditory cells has not yet been demonstrated. Herein our data clearly demonstrated that cisplatin decreased the viability of HEI-OC1 auditory cells, which was inhibited by the addition of neutralizing $anti-TNF-{\alpha}$, $anti-IL-1{\beta}$ and anti-IL-6 antibodies. Consistently, Neutralization with antibodies against pro-inflammatory cytokines ameliorated the cell death and disarrangement of cochlea hair cell layers in the rat primary cochlear explants which were treated with cisplatin. Furthermore, exogeneous supplementation with free radical scavengers, including GSH and NAC, significantly prevented the cytotoxicity of cisplatin in the rat primary cochlea explants. We also observed that $TNF-{\alpha}$ was predominantly expressed in Deiters and Hensen's cells located in hair cell zone of cisplatin-treated cochlear explants. These findings suggest that pro-inflammatory cytokines, including $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6, may play a pivotal role in the pathophysiology of hair cell damages caused by ototoxic drug cisplatin.

Comprehensive investigations of key mitochondrial metabolic changes in senescent human fibroblasts

  • Ghneim, Hazem K.;Alfhili, Mohammad A.;Alharbi, Sami O.;Alhusayni, Shady M.;Abudawood, Manal;Aljaser, Feda S.;Al-Sheikh, Yazeed A.
    • The Korean Journal of Physiology and Pharmacology
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    • v.26 no.4
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    • pp.263-275
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    • 2022
  • There is a paucity of detailed data related to the effect of senescence on the mitochondrial antioxidant capacity and redox state of senescent human cells. Activities of TCA cycle enzymes, respiratory chain complexes, hydrogen peroxide (H2O2), superoxide anions (SA), lipid peroxides (LPO), protein carbonyl content (PCC), thioredoxin reductase 2 (TrxR2), superoxide dismutase 2 (SOD2), glutathione peroxidase 1 (GPx1), glutathione reductase (GR), reduced glutathione (GSH), and oxidized glutathione (GSSG), along with levels of nicotinamide cofactors and ATP content were measured in young and senescent human foreskin fibroblasts. Primary and senescent cultures were biochemically identified by monitoring the augmented cellular activities of key glycolytic enzymes including phosphofructokinase, lactate dehydrogenase, and glycogen phosphorylase, and accumulation of H2O2, SA, LPO, PCC, and GSSG. Citrate synthase, aconitase, α-ketoglutarate dehydrogenase, succinate dehydrogenase, malate dehydrogenase, isocitrate dehydrogenase, and complex I-III, II-III, and IV activities were significantly diminished in P25 and P35 cells compared to P5 cells. This was accompanied by significant accumulation of mitochondrial H2O2, SA, LPO, and PCC, along with increased transcriptional and enzymatic activities of TrxR2, SOD2, GPx1, and GR. Notably, the GSH/GSSG ratio was significantly reduced whereas NAD+/NADH and NADP+/NADPH ratios were significantly elevated. Metabolic exhaustion was also evident in senescent cells underscored by the severely diminished ATP/ADP ratio. Profound oxidative stress may contribute, at least in part, to senescence pointing at a potential protective role of antioxidants in aging-associated disease.

EFFECTS OF HYDROQUINONE ON NEOPLASTIC TRANSFORMATION OF HUMAN EPITHELIAL CELLS IN CULTURE (Hydroquinone이 인체 상피세포의 발암화에 미치는 영향)

  • Sohn, Jung-Hee;Kim, Chin-Soo
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.32 no.3
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    • pp.218-228
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    • 2010
  • Components of dental resin-based restorative materials are reported to leach from the filling materials even after polymerization. Hydroquinone (HQ) is one of the major monomers used in the dental resin and is known as a carcinogen. Thus, carcinogenic risk of HQ leaching from the dental resin becomes a public health concern. The present study attempted to examine the carcinogenic potentials of HQ on the human epithelial cell, which is the target cell origin of the most of oral cancers. Cytotoxicity of HQ was observed above 50${\mu}M$ as measured by LDH assay, indicating a relatively low toxicity of this substance in human epithelial cells. The parameters of neoplastic cellular transformation such as cell saturation density, soft agar colony formation and cell aggregation were analyzed to examine the carcinogenic potential of HQ. The study showed that 2-week exposure of HQ showed the tendency of increase in the saturation density and the significant enhancement of soft agar colony formation at the highest dose, 50 ${\mu}M$ only. It is suggested that HQ has a weak potential of carcinogenicity. When cells were treated with HQ and TPA, a well-known tumor promoter, the parameters of neoplastic cellular transformation was significantly increased. This result indicates that the potential risk of carcinogenicity from HQ is largely dependent upon the presence of promoter. Exposure of 50 ${\mu}M$ HQ increased the time-dependent apoptosis as measured by the ELISA kit. This concentration coincides with a dose of neoplastic transformation, indicating a possible link between apoptosis and HQ-induced cellular transformation. Hydroquinone generated Reactive Oxygen Species (ROS) which was evidenced by the treatment of antioxidants such as trolox and N-acetyl cysteine and the GSH depleting agent, BSO. Antioxidants blocked the generation of ROS and the GSH depleting agent, BSO dramatically increased the ROS production. Since HQ is known to increase ROS production thru activation of transcriptional factor such as c-Myb and Pim-1, it is speculated that ROS generation by HQ plays a role in the activation of oncogene, which may lead to neoplastic transformation. In addition, ROS is involved in the alteration of signal transduction, which regulates the apoptosis in many cellular systems. Thus, ROS-mediated apoptosis may be involved in the HQ-induced carcinogenic processes. Protein kinase C (PKC) is known to play pivotal roles in neoplastic transformation of cells and its high expression is often found in a variety of types of tumors including oral cancer. PKC translocation of PKC-${\alpha}$ was observed following HQ exposure. Altered signaling system may also play a role in the transformation process. Taken together, HQ leached from the dental resin does not pose a significant threat as a cancer causing agent, but its carcinogenic potential can be significantly elevated in the presence of promoter. The mechanism of HQ-induced carcinogenesis involved ROS generation, apoptosis and altered signaling pathway. The present study will provide a valuable data to estimate the potential risk of HQ as a carcinogen and understand mechanism of HQ-induced carcinogenesis in human epithelial cells.

Effects of Copper and Selenium Supplementation on Performance and Lipid Metabolism in Confined Brangus Bulls

  • Netto, Arlindo Saran;Zanetti, Marcus Antonio;Claro, Gustavo Ribeiro Del;de Melo, Mariza Pires;Vilela, Flavio Garcia;Correa, Lisia Bertonha
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.4
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    • pp.488-494
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    • 2014
  • Twenty-eight Brangus cattle were used to determine the effect of copper and selenium supplementation on performance, feed efficiency, composition of fatty acids in Longissimus dorsi (LD) muscle, and cholesterol concentration in serum and in LD muscle and enzymes activities, reduced glutathione (GSH) and oxidized glutathione (GSSG). The treatments were: i) Control, without copper (Cu) and selenium (Se) supplementation; ii) Se, 2 mg Se/kg of dry matter such as sodium selenite; iii) Cu, 40 mg Cu/kg of dry matter such as copper sulfate; iv) Se/Cu, 2 mg Se/kg of dry matter such as sodium selenite and 40 mg Cu/kg of dry matter such as copper sulfate. LD muscle fatty acid composition was not influenced by the treatments (p>0.05). The serum concentration of cholesterol was not influenced by the treatments (p>0.05), however, the concentration of cholesterol in LD was lower in cattle supplemented with copper and selenium (p<0.05). Oxidized glutathione and reduced glutathione increased (p<0.05) with Cu, Se and Se/Cu supplementation. The supplementation of copper (40 mg/kg DM) and selenium (2 mg/kg DM) altered the metabolism of lipids in confined Brangus cattle, through a decrease in cholesterol deposition in the LD, possibly by changing the ratio between reduced glutathione/oxidized glutathione. Copper and selenium supplementation improved animal performance and feed efficiency (p<0.05) when compared to the control group, providing advantages in the production system, while also benefiting consumers by reducing cholesterol concentration in the meat.

Antioxidant Activity and Inhibition of MMP-9 by Isorhamnetin and Quercetin 3-O-$\beta$-D-Glucopyranosides Isolated from Salicornia herbacea in HT1080 Cells

  • Kong, Chang-Suk;Kim, You-Ah;Kim, Moon-Moo;Park, Jin-Sook;Kim, Se-Kwon;Lee, Burm-Jong;Nam, Taek-Jeong;Seo, Young-Wan
    • Food Science and Biotechnology
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    • v.17 no.5
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    • pp.983-989
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    • 2008
  • Two flavonoids, isorhamnetin 3-O-$\beta$-D-glucopyranoside (1) and quercetin 3-O-$\beta$-D-glucopyranoside (2), from slander glasswort (Salicornia herbacea, Korean name hamcho) were isolated. Antioxidative and matrix metalloproteinase-9(MMP-9) inhibitory effects of these compounds were investigated in HT 1080 cell lines. These compounds suppressed the electron spin resonance (ESR) signal intensity on generation of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical in a free-cellular system. Their scavenging effects on generation of intercellular reactive oxygen species (ROS) also exhibited similar trends with DPPH radical in the free cellular system. Also, a control group combined only with Fe(II)-$H_{2}O_2$ resulted in DNA apoptosis by oxidative stress, whereas treatments with these compounds suppressed radical-mediated DNA damage. Intracellular glutathione (GSH) levels were slightly increased in the presence of compound 1 and 2. Moreover, these compounds led to the reduction of the expression levels of MMP-9 without cytotoxic influence. These results suggest that these compounds have a potential as a valuable natural antioxidant and MMP inhibitor related to oxidative stress. Therefore, these compounds not only can be developed as a candidate for a therapeutic potential but also a source for use as ingredients of health foods or functional foods to prevent metastasis involving MMP-9, closely related to ROS.

Hepatoprotective Effect of Ethanol Extract of xBrassicaraphanus on Liver Injury in Rats Treated by D-galactosamine (배무채 에탄올층의 D-galactosamine 간손상에 대한 보호효과)

  • Rhee, Yun-Hee;Lee, Eun-ok;Lee, Hyo-Jung;Shim, Bum-Sang;Ahn, Kyoo-Seok;Choi, Jong-Won;Lee, Soo-Seong;Yoon, Byong-Su;Kim, Sung-Hoon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.6
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    • pp.1388-1393
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    • 2007
  • The protective effect of xBrassicoraphanus (BR) on liver inhury was evaluated in the rats with liver injury induced by i.p. injection of D-galactosamine (GalN) following 2 week oral treatment of ethanol extract of xBrassicoraphanus (EBR). EBR (200 mg/kg) significantly suppressed the levels of ALT, AST, SDH, ${\gamma}-GT$, ALP, LDH and lipid peroxidation compared with GalN treated control, while EBR at 100 mg/kg significantly suppressed AST and ${\gamma}-GT$. Similarly, EBR at 200 mg/kg significantly attenuated the levels of Phase I enzymes such as XO, AO, AH and AD as well as significantly increased the levels of Phase II enzymes such as SOD, catalase and GSH-Px in the GalN treated rats. Taken together, these results indicate that the ethanol extract of xBrassicoraphanus may have a hepatoprotective effect against GalN induced liver injury, suggesting the ethanol extract of xBrassicoraphanus can be applied as hepatoprotective functional food. However, its mechanism should be further studied in molecular and cellular view points.

Supplementation with Selenium and Vitamin E Improves Milk Fat Depression and Fatty Acid Composition in Dairy Cows Fed Fat Diet

  • Liu, Zhao L.;Yang, De P.;Chen, Pu;Dong, Wei X.;Wang, Dong M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.6
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    • pp.838-844
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    • 2008
  • This experiment was conducted to evaluate the effect of supplementing a fat diet with selenium (Se) and vitamin E on performance of cows, blood antioxidant status and milk fatty acid composition. Sixty-three lactating Holstein cows were randomly divided into seven groups of nine cows each and each group was fed one of the following diets: i) a basal diet (control); ii) a basal diet with 0.15 mg Se/kg DM (LSe); iii) a basal diet with 0.3 mg Se/kg DM (HSe); iv) a basal diet with 5,000 IU/cow d vitamin E (LVE); v) a basal diet with 10,000 IU/cow d vitamin E (HVE); vi) a basal diet with 0.15 mg Se/kg DM and 5,000 IU/cow d vitamin E (LSeVE); vii) a basal diet with 0.3 mg Se/kg DM and 10,000 IU/cow d vitamin E (HSeVE). Milk fat percentage and conjugated linoleic acid (CLA) yield in HVE and HSeVE diets increased (p<0.05) compared with the control diet. In milk fat, dietary supplementation of Se tended to increase the proportion of the sum of unsaturated fatty acids (UFA) and significantly decreased (p<0.05) the proportion of the sum of saturated fatty acids (SFA). In addition, compared with the control, thiobarbituric acid reactive substances (TBARS) content was lower and glutathione peroxidase (GSH-Px) was higher when fat diets were supplemented with Se. Our data showed that supplementation with Se and/or VE improved these nutrients in blood and milk. The results indicated that fat diets supplemented with Se improved both antioxidant status in blood and fatty acids in milk fat, and fat diets supplemented with vitamin E alleviated milk fat depression. Therefore, the combination of Se and vitamin E caused synergistic effects on the nutritional quality of milk fat and performance of cows fed a fat diet.

Effects of Guemengja (Rosae Laevigatae Michx.) Extracts on Serum Lipid Composition and Antioxidative Capacity in Rats Fed High Oxidized Fat (금앵자(金櫻子) 추출물이 과산화지질을 급여한 흰쥐의 혈청지질구성과 할산화능에 미치는 영향)

  • 박영훈;이은;임상철
    • Korean Journal of Plant Resources
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    • v.17 no.3
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    • pp.323-330
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    • 2004
  • Effects of Guemengja (Rosae Laevigatae Michx.) Extracts on serum and liver lipid composition and antioxidative capacity were investigated in rat fed high oxidized fat. Twenty eight male Sprague-Dawley rats weighing 159.35$\pm$2.17g were blocked into four groups according to body weight and raised seven weeks with basal diet (normal group, I), basal diet and 10% oxidized fat (control group, II), basal diet, 10% oxidized fat and 100mg/kgg Guemengja Extracts (100mg/kg Guemengja Extracts group, III) and basal diet, 10% oxidized fat and 200mg/kg Guemengja Extracts (200mg/kg Guemengja Extracts group, IV). The level of plasma total cholesterol and triglyceride showed a tendency to decrease, whereas the plasma HDL-cholesterol concentration revealed a tendency to increase in guemengja extracts groups. The level of liver total cholesterol showed no significantly different in all treatment groups, however the level of liver triglyceride showed a tendance to decrease in guemengja extracts groups. Thiobarbituric acid(TBARS) values in plasma and liver showed a tendence to decrease in guemengja extracts groups. The guemengja extracts samples have also decreased the plasma GOT and GPT activities, whereas they have increased the liver glutathione peroxidase, superoxide dismutase and catalase activity.