• Title/Summary/Keyword: cytoplasmic effect

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Apoptotic Signaling Cascade of 5-aminolaevulinic Acid-based Photodynamic Therapy in Human Promyelocytic Leukemia HL-60 Cells

  • Nagao, Tomokazu;Matsuzaki, Kazuki;Takahashi, Miho;Minamitani, Haruyuki
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.509-511
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    • 2002
  • In this study, we investigated apoptotic cell death induced by photodynamic therapy using 5-aminolaevulinic acid (ALA-PDT) in human promyelocytic leukemia cells (HL-60). ALA-PDT induced apoptosis in HL-60 cells as confirmed by DNA agarose gel electrophoresis and nuclear staining with Hoechst 33342. The apoptotic cell death was inhibited by addition of broad-spectrum caspase inhibitor Z-Asp-CH$_2$-DCB, indicating that the apoptotic cell death was induced in a caspase-dependent manner. Actually, western blotting analysis revealed that caspase-3 was processed as early as 1.5 h after ALA-PDT. Cytoplasmic cytochrome c released from mitochondria was detected by western blotting. However, inhibitor of caspase-9, a cysteine protease located in the downstream of cytochrome c release, was not able to reduce the apoptotic cell death. Therefore, the mitochondrial apoptotic pathway was not involved in the ALA-PDT-induced apoptosis. On the other hand, it was found that ALA-PDT-induced apoptosis was clearly inhibited by pretreatment of caspase-8 inhibitor. These data suggest that caspase-8-mediated apoptotic pathway is important in ALA-PDT-induced cell death.

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Decreased Expression of Aquaporin-2 Water Channels in the Kidney in Rats Treated with Reserpine

  • Lee, Jong-Un;Oh, Yoon-Wha;Kim, Sun-Mi;Kim, Soo-Wan
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.4
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    • pp.307-313
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    • 2001
  • Whether there exists a sympathetic neural mechanism regulating the expression of aquaporin (AQP) water channels in the kidney was investigated. Male Sprague-Dawley rats were treated with reserpine (1 mg/kg, IP), and the expression of AQP1-4 proteins was determined in the kidney one day thereafter. Following the treatment with reserpine, the systolic blood pressure measured in a conscious state was significantly decreased in the experimental group compared with that in the control $(83{\pm}8\;vs\;124{\pm}6\;mmHg;\;n=6\;each,\;P<0.05)$. The expression of AQP2 proteins was decreased in the cortex, outer medulla, and inner medulla. The decrease of AQP2 proteins was in parallel in the membrane and the cytoplasmic fractions, suggesting a preserved AQP2 targeting. No significant changes were observed in the expression of AQP1, AQP3, or AQP4. Neither basal nor AVP-stimulated formation of cAMP was significantly altered. These results suggest that the sympathetic nervous system has a tonic stimulatory effect specifically on the expression of AQP2 water channels in the kidney.

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Momordica charantia Protects against Cytokine-induced Apoptosis in Pancreatic $\beta$-Cells

  • Kim, Kyong;Kim, Hye-Young
    • Food Science and Biotechnology
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    • v.17 no.5
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    • pp.947-952
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    • 2008
  • The unripe fruit of Momordica charantia (MC) has been shown to possess antidiabetic activity. However, the mechanism of its antidiabetic action has not been fully understood. In this study, the effects of the aqueous ethanolic extract of MC (AEE-MC) were evaluated on the apoptosis in pancreatic $\beta$-cells treated with a combination of the cytokines, interleukin (IL)-$1{\beta}$, tumor necrosis factor (TNF)-$\alpha$, and interferon (IFN)-$\gamma$. In MIN6N8 cells, the inhibitory effect of AEE-MC was significantly observed at 2 to 50 ${\mu}g/mL$: a 26.2 to 55.6% decrease of cytoplasmic DNA fragments quantified by an immunoassay. The molecular mechanisms, by which AEE-MC inhibited $\beta$-cell apoptosis, appeared to involve the inhibition on the expression of p21, Bax, and Bad, the up-regulation of Bcl-2 and Bcl-$X_L$, and the inhibition on the cleavage of caspase-9, -7, and -3 and poly (ADP-ribose) polymerase. This study suggests that MC may inhibit cytokine-induced apoptosis in $\beta$-cells and, thus, may contribute via this action to the antidiabetic influence in diabetes.

Expression of Folate Receptor Protein in CHO Cell Line

  • Kim, Chong-Ho;Park, Seung-Taeck
    • Biomedical Science Letters
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    • v.14 no.4
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    • pp.203-210
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    • 2008
  • One of cell surface receptor proteins, human folate receptor (hFR) involves in the uptake of folates through cell membrane into cytoplasm, and is anchored to the plasma membrane by a fatty acid linkage, which has been identified in some cells as a glycosylphosphatidylinositol (GPI)-tailed protein with a molecular mass of about 40 kDa. The hFR is released by phosphatidylinositol phospholipase C (PI-PLC) because it contains fatty acids and inositol on the GPI tail. Caveolin decorates the cytoplasmic surface of caveolae and has been proposed to have a structural role in maintaining caveolae. It is unknown whether caveolin is involved in targeting, and is necessary for the function of GPI-tailed proteins. To compare the ability of folic acid binding, internalization and expression of hFR, and the effect of caveolin at the both apical and basolateral side of cell surfaces in Chinese hamster ovary (CHO) clone cells overexpressed the hFR and/or caveolin. Our present results suggest a possibility that the overexpression of caveolin does not be involved in expression of hFR, but plays a role as a factor in PI-PLC releasing kinetics, and for a regulation of formation, processing and function of hFR in CHO clone cells overexpressed cavcolin.

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Inhibitory Effect of $Mg^{2+}$ on the Release of $Ca^{2+}$ from Ryanodine Receptor of the Sarcoplasmic Reticulum in the Skeletal Muscle (골격근 망상체 $Ca^{2+}$유리 Channel[Raynodine receptor]의 $Mg^{2+}$에 의한 유리 억제)

  • 이철주
    • Journal of Chest Surgery
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    • v.25 no.4
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    • pp.347-355
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    • 1992
  • The precise mechanism of the Excitation-Contraction Coupling is still uncertain. But the concept that Ca2+ induced Ca2+ release [CICR] from the Ryanodine receptor in the sarcoplasmic reticulum [foot structure] may play a major role in E-C coupling has been widely accepted since 1970`s. It is believed that increased cytosolic Ca2+ followed by CICR is main contributor for E-C coupling of striated muscle. Resulting phenomena of ischemic /post-reperfusion myocyte is increased cytosolic Ca2+, even to the absence of Ca2+ in reperfusate. So intracellular inhibitor to CICR might prevent the ischemic and reperfusion damage of myocardial cells. The relatively purified foot protein, especially heavy sarcoplasmic reticulum rich, of the skeletal muscle was incorporated into the black lipid bilayer [Phosphatidyl ethanolamine: Phosphatidyl serine=l: 1]. Under the steady state of membrane potential [+20 mV], ionic current through Ryanodine receptor was measured with Cs+ as charge carrier. In the cis chamber [Cytoplasmic side], Mg2+ strongly inhibited CICR of Ryanodine receptor[Kd=6.2 nM]. In conclusion, naturally existing intracellular free Mg2+ can inhibit CICR from intracellular Ca2+ reservior [heavy SR]. So post-ischemic or post-reperfusing myocardium could be preserved using additional free Mg2+ in cardioplegic solution or reperfusate, otherwise the optimal concentration is undetermined.

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Experimental Study on the Effects of GamiSamgieum (SGMX) on Hyperlipidemia

  • Kim, Hun;An, Joung-Jo;Jo, Hyun-Kyung;Yoo, Ho-Rhyong;Seol, In-Chan;Kim, Yoon-Sik
    • The Journal of Korean Medicine
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    • v.30 no.3
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    • pp.86-97
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    • 2009
  • Objectives : This study was aimed to investigate the effects of GamiSamgieum (SGMX) on hyperlipidemia using an animal model. Additionally, the underlying mechanisms were investigated by determination of gene expressions related with lipid metabolism. Methods : Forty mice were selected for use in this experiment, and then divided into five groups; Naive, Induced, SGMX 100, SGMX 200, and Lovastatin group as positive control with 8 mice each, and their blood was collected for analysis of blood and serum parameters. Results : Administration of SGMX significantly inhibited the increase of liver weight and the macrovacuolar cytoplasmic alterations in histopathologic finding. SGMX administration significantly protected the liver from pathologic elevation of AST and ALT and from lipid peroxidation. SGMX administration significantly lowered total cholesterol levels and TG, and partially upregulated the gene expression of LCAT, CYP7A1 and LDL-R receptor. Conclusion : SGMX is suggested to possess hypolipidemic effect, and thus could be a potential candidate for herbal hypolipidemic via further studies.

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Estrogen Effect on the Na,K-ATPase Activity Repressed by IgE-Dependent Histamine-Releasing factor in HeLa Cells (HeLa 세포에서 Estrogen이 Na,K-ATPase활성에 미치는 영향)

  • Lee Si-Nae;Kim Hwa-Jung;Lee Kyunglim
    • Microbiology and Biotechnology Letters
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    • v.33 no.4
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    • pp.322-326
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    • 2005
  • IgE-dependent histamine-releasing factor (HRF) is found extracellularly to regulate the degranulation process of histamine in mast cells and basophils and known to play a predominant role in the pathogenesis of chronic allergic disease. HRF has been also identified in the intracellular region of the cell. Previously, we reported that HRF interacts with the 3rd cytoplasmic domain of the alpha subunit of Na,K-ATPase and inhibits Na,K-ATPase activity. Since it is known that estroaen activates the sarcolemmal Na,K-ATPase, we tested whether estrogen recovers the Na,K-ATPase activity repressed by HRF. In this study, we showed that estrogen activates Na,K-ATPase repressed by HRF. RT-PCR and western blot analysis showed that estrogen doesn't reduce the expression level of HRF in HeLa cell, suggesting that this recovery effects of estrogen probably occur via indirect mechanism on HRF and Na,K-ATPase.

Effects of Alkylating Agent on the Sertoli Cell of the Seminiferous Tubule in the Mouse (Alkylating agent가 생쥐 정소의 Sertoli Cell에 미치는 영향)

  • Jung, Hae-Man;Cho, Kwang-Phil;Kim, Jeong-Sang
    • Applied Microscopy
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    • v.26 no.3
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    • pp.293-303
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    • 1996
  • This paper aims to probe that the effect of high dose of cyclophosphamide to the Sertoli cells of the mouse was examined by transmission electron microscope. In the normal group, Sertoli cells contact each other around the basal aspect of the seminiferous tubule, forming numerous row of tight junction, blood-testis barrier. Sertoli cells contain smooth endoplasmic reticulum, well developed Golgi comples, a number of round mitochondria and microfilament. The cytoplasmic necrosis are observed from the 1-time treated group. In the 2-times treated group, smooth endoplasmic reticulum are more developed than normal group, but cisternae are partially dilated. In the 3-times treated group, the smooth endoplasmic reticulum are not developed. In the 2-times treated group, the inner membrane of the mitochondria are partially disrupted, and cristae are all disrupted in the 3-times treated group. The microfilaments are not observed in the all treated groups. According to the results above, it seems that smooth endoplasmic reticulum, mitochondria, and microfilament are disrupted by toxic effects of the cyclosphamide to the Sertoli cells of the mouse.

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Selective Antimicrobial Effects of Spice Extracts Against Lactobacillus plantarum and Leuconostoc mesenteroides Isolated from Kimchi. (김치에서 분리한 Lactobacillus plantarum과 Leuconostoc mesenteroides에 대한 향신료 에탄올추출물의 선택적 항균효과)

  • 김옥미;김미경;이갑랑;김순동
    • Microbiology and Biotechnology Letters
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    • v.26 no.5
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    • pp.373-378
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    • 1998
  • This study was performed to investigate antimicrobial effects and their actions of the ethanol extracts of 12 spices against Lactobacillus plantarum and Leuconostoc mesenteroides, which are related to Kimchi fermentation. The cardamon, thyme, and cumin ethanol extracts among 12 spices showed antimicrobial activities against Lac. plantarum, not against Leu. mesenteroides through paper disc method. The growth of Lac. plantarum was inhibited in MRS broth containing each extracts of cardamon (>4%), thyme (>4%) and cumin (>1%). SDS-PAGE and transmission electron micrographs showed that the cell walls and membranes were disrupted and the cytoplasmic components were leaked in strains treated with ethanol extracts.

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Ginseng total saponin modulates podocyte p130Cas in diabetic condition

  • Ha, Tae-Sun;Lee, Jin-Seok;Choi, Ji-Young;Park, Hye-Young
    • Journal of Ginseng Research
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    • v.37 no.1
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    • pp.94-99
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    • 2013
  • Proteinuric conditions demonstrate structural and compositional changes of the foot processes and slit diaphragms between podocytes. p130Cas in podocytes serves as an adapter protein anchoring glomerular basement membrane to actin filaments of podocyte cytoskeleton. To investigate the effect of ginseng total saponin (GTS) on the pathologic changes of podocyte p130Cas induced by diabetic conditions, we cultured mouse podocytes under: 1) normal glucose (5 mM, control); 2) high glucose (HG, 30 mM); 3) advanced glycosylation endproducts (AGE)-added; or 4) HG plus AGE-added conditions and treated with GTS. In confocal imaging, p130Cas colocalized with zonula occludens-1 and synaptopodin connecting to F-actin. However, diabetic conditions relocalized p130Cas molecules at perinuclear cytoplasmic area and reduced the intensity of p130Cas. In Western blotting, diabetic conditions, especially HG plus AGE-added condition, decreased cellular p130Cas protein levels at 24 and 48 h. GTS improved such quantitative and qualitative changes. These findings imply that HG and AGE have an influence on the redistribution and amount of p130Cas of podocytes, which can be reversed by GTS.