Functional analysis of nitric oxide synthase related genes transfered into cardiomyocytes in viv

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MethodsEndothelial 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.Results1)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.DiscussionsecNOS 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.