1997 Fiscal Year Final Research Report Summary
Functional analysis of nitric oxide synthase related genes transfered into cardiomyocytes in viv
| Project/Area Number |
08670761
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Circulatory organs internal medicine
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| Research Institution | The University of Tokyo |
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Principal Investigator |
SHIN Wea.Soo Univ.of Tokyo, Health Service, instructer, 保健管理センター, 助手 (10211971)
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| Co-Investigator(Kenkyū-buntansha) |
SAKAMOTO Aiji Cellular Physiology Laboratory, RIKEN,The Special Postductoral Researcher of RIK, 理科学研究所, 基礎科学研究員 (40291067)
TOYOOKA Teruhiko University of Tokyo, Faculty of Medicine (Hospital), Professor, 医学部, 教授 (00146151)
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| Project Period (FY) |
1996 – 1997
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| Keywords | nitric oxide synthase / gene transfered / virus vector / cardiomyocytes / apoptosis / beta-galactosidase / NOS inhibitors / mitochondrial |
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| Research Abstract |
Methods
Endothelial nitric oxide synthase(ecNOS)gene has been tranafered into in vivo rat cardiomyocytes with combination of HVJ(Hemagglutinating virus of Japan)and liposomes to examine the direct of nitric oxide in a living heart.
Results
1)The expression of the marker gene, beta-gal(beta-galactosidase)gene was detected 3 days after the gene transfer, reached the maximal level, and still remained 14 days after the in vivo transfer. The efficiency of the gene transfer without HVJ-loposome was less than 10%. The beta-gal gene transfer with HVJ-loposome method did not induce inflammatory findings, and the extent of the gene localized within a intercalated disc which enabled us to distinguish the cellular change by gene transfer from untransfected parts of the same cell.
ecNOS gene transfer into a living geart induced severe tissue damage, unexpectedly. The cardiac tissue damage co-localized with beta-gal and ecNOS transfected area. The gene transfer of less amount(5mug vs.50mug)evoked milder tissue injury. Pretreatment of ^<L->NAME,a NOS inhibitor, suppressed the tissue damage by ecNOS transfection. The area of tissue damage contained accumulation of macrophages and fibrosis with a few area of TUNEL-positive cells. Electronmicroscope showed enlarged and deformed mitochondrial accumulation beside disarray of sarcomeres.
Discussions
ecNOS gene transfer into a living heart which was expected to protect heart from tissue injury induced severe cardiac damage. Influence of the introduced NOS amount or of surrounding condition has been analyzed.
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