| Co-Investigator(Kenkyū-buntansha) |
TAKEBAYASHI Shigeo Fukuoka Univ., School of Medicine, Professor, 医学部, 教授 (80078740)
OHTA Takao Univ. of the Ryukyus, Fac.Med., Professor, 医学部, 教授 (70185271)
SAKU Keijiro Fukuoka Univ., School of Medicine, Professor, 医学部, 教授 (40183371)
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| Budget Amount *help |
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2000: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1999: ¥800,000 (Direct Cost: ¥800,000)
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| Research Abstract |
We performed this project using a new gene delivery system with peptide of N,N-Dipalmitylglycyl-apolipoprotein E motif (129-169)(dpGapoE). First, we examined possibility of this gene delivery system in the lipid metabolism in vitro. Antisense treatment against human cholesteryl ester transfer protein (hCETP) was performed in hCETP transfected cell line. After transfection, translocation of antisense was detected in the cells, especially in the lysosome and the nucleus. Cellular CETP mRNA levels and its activity significantly declined. When LDL added to culture media of cultured cells before transfection with antisense and dpGapoE complex, reduction of transfection was noted. We conclude that, in vitro, dpGapoE effectively deliver antisense into cells through apoE receptor, and it can react with DNA to prevent protein transcription. We applied this method to organ culture and animal model. In liver organ culture, similar results were obtained. However, in animal model, when dpGapoE and
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antisense complex was administered via portal vein, most of them was trapped in the Kupffer cells and endothelial cells in the liver, while few hepatocytes near the portal area showed to be translocation. To examine possibility of gene therapy for prevention of atherosclerosis, we next adopted human CETP transgenic (CETP-Tg) mice, since they were characterized by decreased HDL-cholesterol (HDL-C) and increased LDL-cholesterol (LDL-C). We gave a cholesterol rich diet to CETP-Tg and wild mice. In cholesterol fed mice, plasma levels of total cholesterol and LDL-C increased, and were significantly greater in wild mice than CETP-Tg. After feeding with cholesterol, HDL-C was still lower in CETP-Tg than wild mice. The extent of lipid rich lesion in the aorta was positively correlated with plasma level of total cholesterol and LDL-C.Its area was, however, significantly smaller in CETP-Tg than wild mice. However, CETP-Tg had greater cholesterol contents in the liver and the feces. These results indicated that cholesterol excretion was quite active in CETP-Tg, represented that receptor mediated pathway, such as SR-BI, could be activated. It is, therefore, difficult to prevent HDL uptake in the liver of this animal to prevent atherosclerosis. Less
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