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

• Project/Area Number: 16590065
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biological pharmacy
• Research Institution: Showa 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: ¥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)
• Keywords: oxidized 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

Research Products

• [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
• [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
• [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
• [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
• [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
• [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
• [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
• [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
• [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
• [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
• [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
• [Journal Article] Oxidized phosphatidylcholine in alveolar macrophages in idiopathic interstitial pneumonias (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 vol.183 Pages: 109-121
  • Description: 「研究成果報告書概要(欧文)」より
• [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
• [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
• [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
• [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
• [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
• [Journal Article] Oxidized phosphatidylcholine in alveolar macrophages in idiopathic interstitial pneumonias (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
• [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
• [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
• [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
• [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
• [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)
• [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)
• [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
• [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
• [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
• [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
• [Journal Article] Oxidized low density lipoprotein levels circulating in plasma and deposited in the tissues : Comparison between Helicobacter pylori associated gastritis and acute myocardial infarction (2004)
  • 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
• [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
• [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
• [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