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Analysis of action mechanism of atrial matri-uretic peptide in perfused heart and tissue-cultured myocytes -in vivo analysis using a muti emission-reflexion analyzer and NMR-

Research Project

Project/Area Number 01480156
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field Pathological medical chemistry
Research InstitutionKagawa Medical School

Principal Investigator

HATASE Osamu  Kagawa Med. Sch., Med. Professor, 医学部, 教授 (50033220)

Co-Investigator(Kenkyū-buntansha) TOKUDA Masaaki  Kagawa Med. Sch., Med. Assistant Professor, 医学部, 助手 (10163974)
ITANO Toshifumi  Kagawa Med. Sch., Med. Assistant Professor, 医学部, 助手 (60145042)
MATSUI Hideki  Kagawa Med. Sch., Med. Associate Professor, 医学部, 助教授 (30157234)
Project Period (FY) 1989 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1991: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1990: ¥1,000,000 (Direct Cost: ¥1,000,000)
KeywordsAtrial Natriuretic Peptide / Mitochondria / Perfused Rat Heart / CPAE Cells / Protein Phosphorylation / Calcium Ion / 心房性利尿ペプチド / 瞬間マルチ測光システム / 初代培養心筋細胞 / 酸化防元反応 / 酸化還元反応 / 虚血心 / 灌流心
Research Abstract

The effect of rANP(rat atrial natriuretic peptide) on mitochondrial respiratory state in perfused rat heart.
Purified mitochondria was used for the assay of oxidative phosphorylation using oxygraph. rANP at the concentration of 10ng/ml did not affect the respiratory function of heart mitochondria in vitro. Perfused rat heart with Langendorf apparatus was then used for the assay of mitochondrial respiratory function. A multi-light-emission-reflection monitoring system was used for this assay, and oxidation-reduction state of cytochromes c, al-a3, b, myoglobin was monitored through the experiment. rANP at 10ng/ml significantly prolonged the transient time from oxidative state to reduced state which was induced by ischemic condition.
This indicates that rANP made heart cells more resistant to ischemic damage through the protection of mitochondrial respiratory function. Electro-microscopic study also supported these results.
The effect of hANP(human atrial natriuretic peptide) on phosphorylation and intracellular concentration of calcium ion of CPAE cells derived from bovine pulmonary artery endothelium.
Cultured CAPE cells were used for the phosphorylation assay under aerobic and anerobic condition using [32P orthophosphate in the culture medium. hANP at a concentration of 10ng/ml was added to the medium and the phosphorylation was analyzed and compared. hANP did not affect the phosphorylation under the aerobic condition. In contrast, hANP recovered to some extent the reduced phosphorylation which occurred under anerobic condition.
The concentration of intracellular calcium ion([Ca i) was monitored by a calcium-specific fluorescent dye, Fura 2, and a calcium imaging analysis was performed. Under the anerobic condition, [Ca i raised by 30-50% as compared to that under aerobic condition. hANP(10ng/ml) reduced the incraese of [Ca i to 20-30%. hANP itself did not stimulate the increase of [Ca i.

Report

(4 results)
  • 1991 Annual Research Report   Final Research Report Summary
  • 1990 Annual Research Report
  • 1989 Annual Research Report
  • Research Products

    (13 results)

All Other

All Publications (13 results)

  • [Publications] S.Etoh: "Purification and immunohistochemical study of actin in mitochondrial matrix." Biochemistry International. 20. 599-606 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] T.Taoka: "Induction of differentiation of HL60 cells by a protein kinase C inhibitor,K252a." Biochemical Biophysical Research Communication. 170. 1151-1156 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] T.Tasaka: "Mechanism of transient increase in intracellular concentration of free calcium ions in HL-60 cell differentiation induced by vitamin-D_3 and phorbol ester." Biochemistry International. 23. 137-143 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] H.Nishio: "The evidence for post-meiotic expression of a testis-specific isoform of a regulatory subunit of calcineurin using a monoclonal antibody." Biochemical Biophysical Research Communication. 187. 828-831 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Seiji Etoh, Hideki Matsui, Masaaki Tokuda, Toshifumi Itano, Mitsuo Nakamura, Osamu Hatase: "Purification and immunohistochemical study of actin in mitochondrial matrix." Biochemistry International. 20(3). 599-606 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Teruhisa Taoka, Masaaki Tokuda, Osamu Hatase, Shozo Irino, Anthiny W. Norman: "Induction of differentiation of HL60 cells by a protein kinase C inhibitor, K252a." Biochem. Biophys. Res. Commun.170. 1151-1156 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Taizo Tasaka, Masaaki Tokuda, Teruhisa Taoka, Toshifumi Itano, Hideki Matsui, Seiji Etoh, Hajime Nishio, Osamu Miyamoto, Shozo Irino, Osamu Hatase: "Mechanism of transient increase in intracellular concentration of free calcium ions in HL-60 cell differentiation induced by vitamin-D_3 and phorbol ester." Biochemistry International. 23(1). 137-143 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Hajime Nishio, Hideki Matsui, Lizomar, P. Moia, Shigeo Taketa, Kazuhiro Miyamoto, Masaaki Tokuda, Toshifumi Itano, Sukita Nakahara, Osamu Hatase: "The evidence for post-meiotic expression of a testis-specific isoform of a regulatory subunit of calcineurin using a monoclonal antibody." Biochem. Biophys. Res. Commun.187(2). 828-831 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Osamu Hstase: "Effect of atrial natriuretic peptide on the production of superoxide and the intracellular pH in cultured myocytes and arterial endothelial cells." Japanese Journal of Physiology.

    • Related Report
      1991 Annual Research Report
  • [Publications] Masaki Tokuda et al.: "Effect of atrial natriuretic peptide(ANP)on oxygen consumption in perfused rat heart." Biophys.Biochem.Res.Commun.

    • Related Report
      1990 Annual Research Report
  • [Publications] Hideki Matsui et al.: "Indentification of two calcineurin isoforms in bovine brain by two different monoclonal antibodies." Biophys.Biochem.Res.Commun.

    • Related Report
      1990 Annual Research Report
  • [Publications] Seiji,Etoh et al.: "Isolation and sequence of novel rat testis cDNA coding for calcineurin βーsubunit,Ca^<2+>ーbinding regulatory subunit of a calmodulin stimulated protein phosphatase." Biophys.Biochem.Res.Commun.

    • Related Report
      1990 Annual Research Report
  • [Publications] Masaaki Tokuda,Toshifumi Itano,Hideki Matsui,Seiji Etoh and Osamu Hatase: "Effect of atrial natriuretic peptide(ANP)on oxygen consumption in perfused rat heart.(投稿予定)" Biophys.Biochem.Res.Commun.

    • Related Report
      1989 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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