| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2003: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2002: ¥2,200,000 (Direct Cost: ¥2,200,000)
|
|---|
| Research Abstract |
1)Gene expression in myocarditis and dilated cardiomyopathy
To investigate gene expression in the myocardium of EAM, absolute copy numbers of 44 mRNA species[calcium-handling proteins, contractile proteins, natriuretic peptides(NPs), cytokines, chemokines, growth factors, renin-angiotensin-aldosterone(RAA) system, endothelins(ETs) and extracellular matrix] in synthesized cDNA from a fixed quantity of total heart RNA were assessed using real-time reverse-transcriptase PCR at days 0,14,21 and 28 after immunization. alpha-Cardiac myosin showed a 26.3-fold decrease and beta-cardiac myosin a 3.75-fold increase at day 14. Atrial NP and brain NP increased 47.7-and 6.35-fold at days 21 and 14 respectively. Angiotensin II type I receptor, angiotensin-converting enzyme and ET1 increased 22.3-fold at day 21,6.30-fold at day 21 and 16,8-fold at day 14 respectively. Aldosterone receptor decreased 2.15-fold at day 14,but aldosterohe synthetase was detected only at days 14 and 21. Interleukin(IL)-2,IL
… More
-10,interferon-gamma and monocyte chemo-attractant protein-1 increased 9.08-fold at day 14,398-fold at day 21,43.1-fold at day 14 and 142-fold at day 14 respectively. Collagen type 3, collagen type 1 and fibronectin increased 34.6-,1.74-and 44.4-fold respectively at day 21. Interestingly, osteopontin showed a 4540-fold increase and it was the highest mRNA of all at day 14. An isofonn of cardiac myosin and NP are dramatically changed in EAM. RAA system and ET expressions are changed differently during the EAM time course. Cytokine, chemokine and extracellular matrix greatly increase and, in particular, large numbers of osteopontin mRNA are expressed in early EAM. After acute inflammation healed, rats were treated with angiotensin converting enzyme inhibitors(ACEI) and type 1 All receptor blockers. These agents had favorable effects on hemodynamics and myocardial contractility, prevented fibrosis, suppressed the expression of ANP, and reversed phenotypic change of cardiac myosin. All receptor blockers were less effective than ACEI.
2)Basic study of gene therapy by naked plasmid
Gene transfer into the liver via rapid tail vein injection can easily be achieved in the rat, which is more than 10 times larger than the mouse, and has significant value for gene function analysis in rats. pCAGGS-vIL-10 gene transfer by hydrodynamics-based transfection suppresses crescentic glomerulonephritis. IFN-gamma, TNF-alpha and MCP-1 to the transcripts of G6PDH in the glomeruli were all significantly lower in the pCAGGS-vIL-10 rats than in the pCAGGS rats. We demonstrated a useful and convenient method to assay gene therapy products circulating in blood using a glucagon 19-29 tagging vector. Less
|
