| Project/Area Number |
06671041
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
内分泌・代謝学
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| Research Institution | KUMAMOTO UNIVERSITY |
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Principal Investigator |
KOBORI Sozo (1995) SCHOOL OF MEDICINE,ASSISTANT PROFESSOR Kumamoto University, 医学部・附属病院, 講師 (00201492)
竹田 晴生 (1994) 熊本大学, 医学部, 講師 (80155019)
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| Co-Investigator(Kenkyū-buntansha) |
MIYATA Takao SCHOOL OF MEDICINE,RESEARCH ASSOCIATE Kumamoto University, 医学部, 助手 (70244118)
小堀 祥三 熊本大学, 医学部・附属病院, 講師 (00201492)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1994: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | Mesangial cells / beta-VLDL / LDL receptor / VLDL receptor / foam cells / β超低密度リポ蛋白 / 腎メサンギウム細胞 / 腹腔マクロファージ / 低密度リポ蛋白受容体 / アセチル化低密度リポ蛋白 / スカベンジャー受容体 / コレステリルエステル / 泡沫細胞 |
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| Research Abstract |
1.Analysis of a mechanism of foam cell formation in cultured rat mesangial cells
To elucidate whether b-migrating very low density lipoproteins (beta-VLDL) induce foam cell formation in mesangial cells or not, surface binding and foam cell formation with beta-VLDL were studied in mouse mesangial cells. Spenfic binding kinetics for beta-VLDL and low density lipoproteins (LDL) on the mesangial cells were observed with Kd=3.8and 13.7mug/ml, and Bmax=65.9 and 71.9ng/mg cell protein at 4ﾟC,respectively. The binding of beta-VLDL was inhibited by excess amount to LDL or beta-VLDL,but not acetyl-low density lipoproteins. Ligand bloting using beta-VLDL or LDL and immunoblotting using anti-human LDL receptor monoclonal antibody detected a same apparent single protein (around 130 kDa). Incorporation of [^<14>C] -oleate into cholesteryl ester in mouse mesangial cells was enhanced by beta-VLDL to 3-fold higher than that by LDL,and it was inhibited by chloroquine or anti-human LDL receptor monoclonal
… More
antibody. The light lighe microscopic findings also demonstrated that cholesteryl ester deposition increased in these cells incubated with beta-VLDL,but not with LDL.
In conclusion, beta-VLDL was specifically taken up by receptor-mediated endocytosis in mouse mesangial cells through LDL receptors, resulting in foam cell formation.
2.Analysis of a mechanism of foam cell formation in cultured low density lipoprotein receptor deficient rabbit mesangial cells
To obtain a VLDL receptor cDNA fragment, RT-PCR was performed on the mRNA prepared from LDL receptor deficient rabbit mesangial cells using the primers designed on the basis of a rabbit VLDL receptor cDNA sequence. The sequence of the cDNA fragment corresponded to that of the vabbit VLDL receptor cDNA and it was used as the probe for the Northern analysis. In the Northern analysis using the probe, we comfirm that there exist VLDL receptor mRNA in LDL receptor deficient rabbit mesangial cells. Intensities of the mRNA band were not changed before and after loading of cholesterol. From this result, it is suggested that beta-VLDL may be taken up through the VLDL receptor into mesangial cells. Less
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