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2007 Fiscal Year Final Research Report Summary

Protection against ischemia-reperfusion injury using a short interfering RNA that inhibits the activity of myeloperoxidase

Research Project

Project/Area Number 18390428
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Anesthesiology/Resuscitation studies
Research InstitutionKyoto University

Principal Investigator

ARAI Toshiyuki  Kyoto University, Medicine, Associate Professor (80175950)

Co-Investigator(Kenkyū-buntansha) YAMASHITA Kouhei  Kyoto University, Medicine, Assistant Professor (80402858)
Project Period (FY) 2006 – 2007
KeywordsNeutreopils / Myeronperoxidaes / RNA interference / Short interfering RNA / Real time PCR / Reactive oxygen species / Human umbilical endothelial cells / Interleukin-8
Research Abstract

In order to investigate the feasibility of protection against ischemia-reperfusion injury (IRI) using a short interfering RNA (siRNA) that inhibits the activity of myeloperoxidase (MPO) of human neutrophils, the following studies were Performed.
The siRNA to diminish the MPO activity was designed and synthesized. Both effective (positive) and ineffective (negative) siRNAs were prepared. Then, the siRNA was electrically transferred into neutrophils using NucleofectorR system. The Suction of the expression of mRNA of MPO by a positive siRNA was successfully performed, which was confirmed using a real-time PCR. However, when the neutrophils were activated by opsonized zymosan, the generation of reactive oxygen species (ROS) was not diminished in the neutrophils incorporated with a positive siRNA compared to that in the neutrophils incorporated with a negative siRNA. That is, the RNA interference was not effective to inhibit the MPO activity of neutrophils.
As an alternative capture, we tried to inhibit the ROS generation in endothelial cells using human umbilical vein endothelial cells (HUVECs). First, it was revealed that some stimulants, such as lipopolysaccaride (LPS), induced the expression of interleukin-8 (IL-8) and that IL-8, in turn, induced ROS generation in HUVECs. The siRNA to diminish the expression of IL-8 was designed and synthesized. Both positive and negative siRNAs were prepared. Then, the siRNA was electrically transferred into HUVECs using NucleofectorR system. The Suction of the expression of mRNA of IL-8 by a positive siRNA was successfully performed, which was confirmed using a real-time PCR. And, when the HUVECs were stimulated by LPS, the generation of ROS was diminished in the HUVECs incorporated with a positive siRNA compared to that in the HUVECs incorporated with a negative siRNA. It was known that the ROS generated by endothelial cells hurt themselves during IRI. So, we can say that protection against WI using a siRNA was achieved.

  • Research Products

    (24 results)

All 2008 2007 2006

All Journal Article (16 results) (of which Peer Reviewed: 8 results) Presentation (8 results)

  • [Journal Article] The radical scavenger edaravone(3-methyl-1-phenyl-2-pyrazolin-5-one)reacts with a pterin derivative and produces a cytotoxic substance that induces intracellular ROS generation and cell death.2008

    • Author(s)
      Arai T, Nonogawa M, Makino K, et. al.
    • Journal Title

      J Pharmacol Exp Ther 324

      Pages: 529-538

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Anticancer photodynamic and non-photodynamic effects of pterin derivatives on a pancreatic cancer cell line.2008

    • Author(s)
      Miyoshi T, Arai T, Nonogawa M, et. al.
    • Journal Title

      J Pharmacol Sci (in press)

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] The radical scavenger edaravone (3-methyl-l-phenyl-2-pyrazolin-5-one) reacts with a pterin derivative and produces a cytotoxic substance that induces intracellular ROS generation and cell death2008

    • Author(s)
      Arai, T., Nonogawa, M., Makino, K., Endo, N., Mori, H., Miyoshi, T., Yamashita, K., Sasada, M., Kakuyama, M., Fukuda, K
    • Journal Title

      J Pharmacol Exp Ther 324

      Pages: 529-538

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Anticancer photodynamic and non-photodynamic effects of pterin derivatives on a pancreatic cancer cell line2008

    • Author(s)
      Miyoshi, T., Arai, T., Nonogawa, M., Makino, K., Mori, H., Yamashita, K., Sasada, M., Uchiyama, T
    • Journal Title

      J Pharmacol Sci (in press)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Gene expression in enhanced apoptosis of human lymphoma U937 cells treated with the combination of different free radical generators and hyperthermia.2007

    • Author(s)
      Wada S, Tabuchi Y, Kondo T, et. al.
    • Journal Title

      Free Radical Res 41

      Pages: 73-81

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Effects of edaravone on singlet oxygen released from activated human neutrophils.2007

    • Author(s)
      Sommani P, Arai T, Yamashita K, et. al.
    • Journal Title

      J Pharmacol Sci 1003

      Pages: 117-120

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Reactive oxygen species generation through NADH oxidation by 6-formylpterin derivatives in the dark.2007

    • Author(s)
      Nonogawa M, Pack SP, Arai T, et. al.
    • Journal Title

      Biochem Biophys Res Commun 35

      Pages: 1105-1110

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Effects of intravenous anesthesia with propofol on regional cortical blood flow and intracranial pressure in surgery for Moyamoya disease.2007

    • Author(s)
      Kikuta K, Takagi Y, Nozaki K, et. al.
    • Journal Title

      Surgical Neurology 68

      Pages: 421-424

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Synthesis of 6-formylpterin nucleoside analogs and their ROS generation activities in the presence of NADH in the dark.2007

    • Author(s)
      Nonogawa M, Pack SP, Arai T, et. al.
    • Journal Title

      Org Biochem Mol 5

      Pages: 3314-3319

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Inhibitory effects of 6-formylpterin on HIF-la protein accumulation.2007

    • Author(s)
      Sommari P, Yamashita K, Miyoshi T, et. al.
    • Journal Title

      Biol Pharm Bull 30

      Pages: 2181-2184

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Gene expression in enhanced apoptosis of human lymphoma U937 cells treated with the combination of different free radical generators and hyperthermia2007

    • Author(s)
      Wada, S., Tabuchi, Y., Kondo, T., Cui, Z-G., Zhao, Q-L., Takasaki, I., Salunga, TL., Ogawa, R., Arai, T., Makino, K., Furuta, I
    • Journal Title

      Free Radical Res 41

      Pages: 73-81

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Effects of edaravone on singlet oxygen released from activated human neutrophils2007

    • Author(s)
      Sommani, P., Arai, T., Yamashita, K., Miyoshi, T., Mori, H., Sasada, M., Makino, K
    • Journal Title

      J Pharmacol Sci 103

      Pages: 117-120

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Reactive oxygen species generation through NADH oxidation by 6-formylpterin derivatives in the dark2007

    • Author(s)
      Nonogawa, M., Pack, SP., Arai, T., Endo, N., Sommani, P., Kodaki, T., Makino, K
    • Journal Title

      Biochem Biophys Res Commun 353

      Pages: 1105-1110

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Effects of intravenous anesthesia with propofol on regional cortical blood flow and intracranial pressure in surgery for Moyamoya disease2007

    • Author(s)
      Kikuta, K., Takagi, Y., Nozaki, K., Yamada, K., Miyamoto, S., Kataoka, H., Arai, T., Hashimoto, N
    • Journal Title

      Surgical Neurology 68

      Pages: 421-424

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Synthesis of 6-formylpterin nucleoside analogs and their ROS generation activities in the presence of NADH in the dark2007

    • Author(s)
      Nonogawa, M., Pack, SP., Arai, T., Endo, N., Sommani, P., Kodaki, T., Kotake, Y., Makino, K
    • Journal Title

      Org Biomol Chem 5

      Pages: 3314-3319

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Inhibitory effects of 6-formylpterin on HIF-la protein accumulation2007

    • Author(s)
      Sommani, P., Yamashita, K., Miyoshi, T., Tsunemine, H., Kodaki, T., Mori, H., Hirota, K., Arai, T., Sasada, M., Makino, K
    • Journal Title

      Biol Pharm Bull 30

      Pages: 2181-2184

    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] 敗血症におけるIL-8の新たな役割の解明-血管内皮細胞の活性酸素産生に対する作用-。2007

    • Author(s)
      三好隆史、山下浩平、荒井俊之、伊藤洋志、内山卓、笹田昌孝
    • Organizer
      第69回日本血液学会総会・第49回日本臨床血液学会合同総会
    • Place of Presentation
      横浜
    • Year and Date
      20071011-13
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] The new role of interleukin-8 in sepsis (Reactive oxygen production from endothelial cells by IL-B)2007

    • Author(s)
      Miyoshi, T., Yamashita, K., Arai, T., Itoh, H., Uchiyama, T., Sasada, M
    • Organizer
      The 69th Annual Meeting of the Japanese Society of Hematology
    • Place of Presentation
      Yokohama
    • Year and Date
      20071011-1013
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] ヒト好中球における6-フォルミルプテリンの新誘導体(6FP-tBu-DMF)の一重項酸素産生に及ぼす影響。2007

    • Author(s)
      森浩子、荒井俊之、福田和彦
    • Organizer
      日本麻酔科学会第54回学術集会
    • Place of Presentation
      札幌
    • Year and Date
      20070531-0602
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Effects of a novel pterin derivative on singlet oxygen production in human neutrophils2007

    • Author(s)
      Mori, H., Arai, T., Fukuda, K
    • Organizer
      The 54th Annual Meeting of Japan Society of Anestheologist
    • Place of Presentation
      Sapporo
    • Year and Date
      20070531-0602
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] 細胞透過性プテリン化合物の光化学反応が誘導する皮膚T細胞性リンパ腫細胞株における細胞死の解析。2006

    • Author(s)
      常峰紘子、山下浩平、荒井俊之、笹田昌孝、内山卓
    • Organizer
      第68回日本血液学会総会・第48回日本臨床血液学会総会
    • Place of Presentation
      福岡
    • Year and Date
      20061006-08
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Photoactivation of pterin derivative induces apoptotic and necrotic cell death on a CTCL cell line2006

    • Author(s)
      Tsunemine, H., Yamashita, K., Arai, T., Sasada, M., Uchiyama, T
    • Organizer
      The 68th Annual Meeting of the Japanese Society of Hematology
    • Place of Presentation
      Fukuoka
    • Year and Date
      20061006-08
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] ヒト好中球における一重項酸素産生におよぼすプテリン誘導体6-フォルミルプテリンの影響。2006

    • Author(s)
      森浩子、荒井俊之、石井久成、福田和彦
    • Organizer
      日本麻酔科学会第53回学術集会
    • Place of Presentation
      神戸
    • Year and Date
      20060601-03
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Effects of 6-formylpterin on singlet oxygen production in human neutrophils2006

    • Author(s)
      Mori, H., Arai, T., Ishii, H., Fukuda, K
    • Organizer
      The 53th Annual Meeting of Japan Society of Anestheologist
    • Place of Presentation
      Kobe
    • Year and Date
      20060601-03
    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2010-02-04  

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