A possible treatment strategy for cardiovascular remodeling by metabolic syndrome-The gene therapy for use of MCP-1 7ND mutated gene-

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Production of the MCP-17ND:Fc chimera cDNA (7ND) and verification of its dominant-negative activit :At first, we use PCR method to create 7ND cDNA by conjugating the amino-terminal deleted mutant of human MCP-1 and the human immunoglobulin Fc chain. Recombinant 7ND protein was obtained by overexpressing 7ND cDNA in 293 cells and we verified the dominant-negative activity of the recombinant 7ND protein on the MCP-1-induced macrophage production of interferon-gamma. In WKY rats and SD rats, 7ND gene transfer expressed 7ND protein in the muscle and significant elevation of circulating 7ND levels was documented by Elisa, having significant musclular damage at the site of injection (up to 1 g plasmid/kg body weight). However, we could not detect significant expression in circulating 7ND levels in 7ND (1 g/kg)-treated OLEFT rats. One explanation is the difference of the model. Another one is the muscle damage at the transfected site. Because OLEFT rats grow substantial larger than WKY and SD … More rats, the net dose of plasmid solution got much larger, resulting in the myocyte lysis and necrosis. Thus, we are now trying to search more efficient vectors.Evaluation of cardiac remodeling at multiple risk factor model (OLETF Rat) :We used OLETF rats to investigate cardiovascular remodeling in diabetes and hyperlipidemia status. LETF rats were use as controls. In stead of 7ND gene therapy, we used a neutralizing antibody against MCP-1 (NAb) to block MCP-1 activity. NAb or control IgG was administered everyday to OLETF and LETF rats from 30 weeks-old. At 40 weeks of age, we evaluated cardiac hypertrophy, fibrosis and cardiac function. In control OLETF rats had been greater interstitial myocardial fibrosis, especially reparative cardiac fibrosis, and sclerotic change of major vessels, compared with control LETF rats. Preceding cardiac fibrosis, macrophage infiltration and fibroblast proliferation were observed in perivascular space in control OLETF rats. This phenomenon suggested that hyperglycemia is involved in both inflammation changes and tissue damage remodeling. NAb treatment failed to reduce macrophage accumulation and cardiac fibrosis in OLETF rat hearts. We speculated that the inhibitory effects of NAb did not last for 10 weeks Less