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Separation and analysis of oxidized low-density lipoprotein in vivo using immunological technique.

Research Project

Project/Area Number 16590065
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Biological pharmacy
Research InstitutionShowa University

Principal Investigator

ITABE Hiroyuki  Showa University, School of Pharmaceutical Sciences, Professor, 薬学部, 教授 (30203079)

Co-Investigator(Kenkyū-buntansha) KATO Rina  Showa University, School of Pharmaceutical Sciences, Research, 薬学部, 助手 (30392400)
OBAMA Takashi  Showa University, School of Pharmaceutical Sciences, Research, 薬学部, 助手 (60395647)
Project Period (FY) 2004 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2005: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2004: ¥1,800,000 (Direct Cost: ¥1,800,000)
Keywordsoxidized LDL / atherosclerosis / monoclonal antibody / apoE-knockout mouse / mass spectrometry / oxidative stress / LDL
Research Abstract

We have proved the occurrence of oxidized low-density lipoprotein (OxLDL) in human circulating plasma through establishing a sensitive method to measure OxLDL using a monoclonal atnibdy (DLH3) recognizing oxidized phosphatidylcholine. OxLDL, known for a risk factor for atherosclerosis, is a mixture of heterogeneously modified particles. The precise nature of OxLDL present in vivo has not yet been elucidated. In this study we tried to separate OxLDL from human plasma and analyze its modified structures. Elucidation of in vivo OxDL itself should be a crucial step for understanding metabolic behavior and clinical significance of OxLDL in vivo.
It was estimated previously, based on the sandwich ELISA using DLH3 antibody, that there is approximately one OxLDL particle in 10000 LDL particles in human plasma. To analyze such minute amount of materials, we introduced ESI-LC/MS/MS technique. Human LDL pre-treated either with acrolein or 4-hydroxynonenal (HNE) was trypsin-digensted, then the samole was subjected to LC/MS/MS analysis. We successfully detected and identified tryptic fragments of apoB containing acrolein-modified lysine or HNE-modified histidine. These typical modified structures are good markers for oxidative modification of proteins, and they are useful for analysis of OxLDL in vivo. We also successfully detected oxidized phospholipids and oxidized cholesterols using MS/MS. It is very likely that effective concentration step of OxLDL from plasma is needed to analyze plasma OxLDL. Separation of OxLDL in plasma using an OxLDL-absorbing dialysis membrane in a preliminary trial, that and identified tryptic fragments of apoB containing acrolein-modified lysine or HNE-modified histidine. modified with

Report

(3 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • Research Products

    (32 results)

All 2006 2005 2004

All Journal Article (31 results) Book (1 results)

  • [Journal Article] ADRP/adipophilin is degraded through proteasome-dependent pathway during regression of lipid-storing cells.2006

    • Author(s)
      Masuda, Y., Itabe, H., Odaki, M., Hama, K., Fujimoto, Y., Mori, M., Sasabe, N., Aoki, J., Arai, H., Takano,T.
    • Journal Title

      Journal of Lipid Research 47

      Pages: 87-98

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Expression of lectin-like oxidized LDL receptor-1 smooth muscle cells after vascular injury.2006

    • Author(s)
      Eto, H., Miyata, M., Kume, N., Minami, M., Itabe, H., Orihara, K., Hamasaki, S., Biro, S., Otsuji, Y., Kita, T., Tei,C.
    • Journal Title

      Biochem Biophys Res Commun. 241

      Pages: 591-598

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Expression of lectin-like oxidized LDL receptor-1 in smooth muscle cells after vascular injury.2006

    • Author(s)
      Eto, H., Miyata, M., Kume, N., Minami, M., Itabe, H., Orihara, K., Hamasaki, S., Biro, S., Otsuji, Y., Kita, T., Tei, C.
    • Journal Title

      Biochem Biophys Res Commun vol.241

      Pages: 591-598

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] ADRP/adipophilin is degraded through the proteasome-dependent pathway during regression of lipid-storing cells.2006

    • Author(s)
      Masuda, Y., Itabe, H., Odaki, M., Hama, K., Fujimoto, Y., Mori, M, Sasabe, N., Aoki, J., Arai, H., Takano, T.
    • Journal Title

      Journal of Lipid Research 48

      Pages: 87-98

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Expression of lectin-like oxidized LDL receptor-1 in smooth muscle cells after vascular injury.2006

    • Author(s)
      Eto, H., Miyata, M., Kume, N., Minami, M., Itabe, H., Orihara, K., Hamasaki, S., Biro, S., Otsuji, Y., Kita, T., Tei, C.
    • Journal Title

      Biochem Biophys Res Commun. 241

      Pages: 591-598

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Role of macrophage and smooth muscle cell apoptosis in association with oxidized low-density lipoprotein in the atherosclerotic development.2005

    • Author(s)
      Akishima Y, Akasaka Y, Ishikawa Y, Lijin Z, Kiguchi H, Ito K, Itabe H, Ishii T
    • Journal Title

      Modern Pathology 18

      Pages: 365-373

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Oxidized phosphatidylcholine in alveolar macrophages in idiopathic interstitial pneumonia.2005

    • Author(s)
      Yoshimi, N., Ikura, Y., Sugama, Y., Ohsawa, M., Hai, E., Kayo, S., Shirai, N., Yamamoto, S., Inoue, Y., Itabe, H., Yoshikawa, J., Ueda,M.
    • Journal Title

      Lung 183

      Pages: 109-121

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] A redox-sensitive pathway is involved in oxidized LDL-induced downregulation of insulin-like growth factor-1 receptor.2005

    • Author(s)
      Higashi, Y., Pen, T., Du, J., Suhkanov, S., Li, Y., Itabe, H., Parthasarathy, S., Delafontaine,P.
    • Journal Title

      Journal of Lipid Research 46

      Pages: 1266-1277

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Inhibition of brain damage by edaravone,a free radical scavenger, can be monitored by plasma biomarkers that detect oxidative and astrocyte damage in patients with acute cerebral infarction.2005

    • Author(s)
      Uno, M., Kitazata, K., Suzue, A., Matsuzaki, K., Harada, M., Itabe, H., Nagahiro,S.
    • Journal Title

      Free Radical Biology and Medicine 39

      Pages: 1109-1116

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Contribution of an imbalance between oxidant-antioxidant sytems to plaque vulnerability in patients with carotid artery stenosis.2005

    • Author(s)
      Uno, M., Kitazato, K.T., Suzue, A., Itabe, H., Hao, L., Nagahiro,S.
    • Journal Title

      Journal of Neurosurgery 103

      Pages: 518-525

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Role of macrophage and smooth muscle cell apoptosis in association with oxidized low-density lipoprotein in the atherosclerotic development.2005

    • Author(s)
      Akishima Y, Akasaka Y, Ishikawa Y, Lijun Z, Kiguchi H, Ito K, Itabe H, Ishii T
    • Journal Title

      Modern Pathology vol.18

      Pages: 365-373

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Oxidized phosphatidylcholine in alveolar macrophages in idiopathic interstitial pneumonias2005

    • Author(s)
      Yoshimi, N., Ikura, Y., Sugama, Y., Ohsawa, M., Hai, E., Kayo, S., Shirai, N., Yamamoto, S., Inoue, Y., Itabe, H., Yoshikawa, J., Ueda, M
    • Journal Title

      Lung vol.183

      Pages: 109-121

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] A Redox-Sensitive Pathway is Involved in Oxidized LDL-Induced Downregulation of Insulin-like Growth Factor-1 Receptor.2005

    • Author(s)
      Higashi, Y., Pen, T., Du, J., Suhkanov, S., Li, Y., Itabe, H., Parthasarathy, S., Delafontaine, P.
    • Journal Title

      Journal of Lipid Research vol.46

      Pages: 1266-1277

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Inhibition of brain damage by edaravone, a free radical scavenger, can be monitored by plasma biomarkers that detect oxidative and astrocyte damage in patients with acute cerebral infarction.2005

    • Author(s)
      Uno, M., Kitazato, K.T., Suzue, A., Matsuzaki, K., Harada, M., Itabe, H., Nagahiro, S.
    • Journal Title

      Free Radical Biology and Medicine vol.39

      Pages: 1109-1116

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Contribution of an imbalance between oxidant-antioxidant sytems to plaque vulnerability in patients with carotid artery stenosis.2005

    • Author(s)
      Uno, M., Kitazato, K.T., Suzue, A., Itabe, H., Hao, L, Nagahiro, S.
    • Journal Title

      Journal of Neurosurgery vol.103

      Pages: 518-525

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] ADRP/adipophilin is degraded through proteasome-dependent pathway during regression of lipid-storing cells.2005

    • Author(s)
      Masuda, Y., Itabe, H., Odaki, M., Hama, K,.Fujimoto, Y., Mori, M, Sasabe, N., Aoki, J., Arai, H, Takano, T.
    • Journal Title

      Journal of Lipid Research vol.47

      Pages: 87-98

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Role of macrophage and smooth muscle cell apoptosis in association with oxidized low-density lipoprotein in the atherosclerotic development.2005

    • Author(s)
      Akishima Y, Akasaka Y, Ishikawa Y, Lijun Z, Kiguchi H, Ito K, Itabe H, Ishii T
    • Journal Title

      Modern Pathology 18

      Pages: 365-373

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Oxidized phosphatidylcholine in alveolar macrophages in idiopathic interstitial pneumonias2005

    • Author(s)
      Yoshimi, N., Ikura, Y., Sugama, Y., Ohsawa, M., Hai, E., Kayo, S., Shirai, N., Yamamoto, S., Inoue, Y., Itabe, H., Yoshikawa, J., Ueda, M
    • Journal Title

      Lung 183

      Pages: 109-121

    • Related Report
      2005 Annual Research Report
  • [Journal Article] A Redox-Sensitive Pathway is Involved in Oxidized LDL- Induced Downregulation of Insulin-like Growth Factor-1 Receptor.2005

    • Author(s)
      Higashi, Y., Pen, T., Du, J., Suhkanov, S., Li, Y., Itabe, H., Parthasarathy, S., Delafontaine, P.
    • Journal Title

      Journal of Lipid Research 46

      Pages: 1266-1277

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Inhibition of brain damage by edaravone, a free radical scavenger, can be monitored by plasma biomarkers that detect oxidative and astrocyte damage in patients with acute cerebral infarction.2005

    • Author(s)
      Uno, M., Kitazato, K.T., Suzue, A., Matsuzaki, K., Harada, M., Itabe, H., Nagahiro, S.
    • Journal Title

      Free Radical Biology and Medicine 39

      Pages: 1109-1116

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Contribution of an imbalance between oxidant-antioxidant systems to plaque vulnerability in patients with carotid artery stenosis.2005

    • Author(s)
      Uno, M., Kitazato, K.T., Suzue, A., Itabe, H., Hao, L., Nagahiro, S.
    • Journal Title

      Journal of Neurosurgery 103

      Pages: 518-525

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Bilirubin from heme oxygenase-1 attenuates vascular endothelial activation and dysfunction against pro-inflammatory stresses.2005

    • Author(s)
      Kawamura, K., Ishikawa, K., Wada, Y., Matsumoto, H., Itabe, H., Kodama, T., Maruyama, Y.
    • Journal Title

      Arterioscler.Thromb.Vasc.Biol. 25

      Pages: 155-160

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Role of macrophage and smooth muscle cell apoptosis in association with oxidized low-density lipoprotein in the atherosclerotic development.2005

    • Author(s)
      Akishima Y, Akasaka Y, Ishikawa Y, Lijun Z, Kiguchi H, Ito K, Itabe H, Ishii T.
    • Journal Title

      Mod.Pathol. (印刷中)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Localization of oxidized phosphatidylcholine in macrophages in idiopathic pulmonary fibrosis.2005

    • Author(s)
      Yoshimi, N., Ikura, Y., Sugama, Y., Ohsawa, M., Hai, E., Kayo, S., Itabe, H., et al.
    • Journal Title

      Lung (印刷中)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] HDL-C levels modify the association between plasma levels of oxidatively modified LDL and coronary artery disease in men.2004

    • Author(s)
      Zhang, B., Bai, H., Liu, R., Itabe, H., Takano, T., Saku, K.
    • Journal Title

      Metabolism 53

      Pages: 423-429

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Identification of the major proteins in the lipid droplet-enriched fraction isolated from the human hepatocyte cell line HuH7.2004

    • Author(s)
      Fujimoto, Y., Itabe, H., Sakai, J., Makita, M., et al.
    • Journal Title

      Biochim.Biophys.Acta 1644

      Pages: 47-59

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Basement membrane-type heparan sulfate proteoglycan (perlecan) and low-density lipoprotein (LDL) co-localized in granulation tissues : a possible pathogenesis of cholesterol granulomas in jaw cysts.2004

    • Author(s)
      Yamazaki, M., Cheng, J., Hao, N., Takagi, R., Jimi, S., Itabe, H., Saku, T.
    • Journal Title

      Oral Pathol.Med. 33

      Pages: 177-184

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Oxidized phospholipids in oxidized low-density lipoprotein reduce the activity of tissue factor pathway inhibitor through association with its carboxy-terminal region.2004

    • Author(s)
      Ohkura.N., Hiraishi, S., Itabe, H., et al.
    • Journal Title

      Antioxidants and Redox Signaling 6

      Pages: 705-712

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Oxidized low density lipoprotein levels circulating in plasma and deposited in the tissues : Comparison between Helicobacter pylori associated gastritis and acute myocardial infarction2004

    • Author(s)
      Kayo, S., Ohsawa, M., Ehara, S., Naruko, T., Ikura, Y., Hai, E., Yoshimi, N., Itabe, H., et al.
    • Journal Title

      American Heart Journal 148

      Pages: 818-825

    • Related Report
      2004 Annual Research Report
  • [Journal Article] 生体内酸化LDL生成、動態と血管2004

    • Author(s)
      板部 洋之
    • Journal Title

      血管医学 5

      Pages: 439-446

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Measurement of oxidized LDL present in human plasma and atherosclerotic lesions.2004

    • Author(s)
      Itabe, H., Ueda, M., Uno, M., Takano, T
    • Journal Title

      International Congress Series 1262

      Pages: 87-90

    • Related Report
      2004 Annual Research Report
  • [Book] 酸化ストレスマーカー2005

    • Author(s)
      板部洋之
    • Total Pages
      296
    • Publisher
      学会出版センター
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary

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Published: 2004-04-01   Modified: 2016-04-21  

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