| Project/Area Number |
11470032
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General medical chemistry
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| Research Institution | University of Tokyo |
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Principal Investigator |
SEYAMA Yousuke University of Tokyo, Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (90010082)
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| Co-Investigator(Kenkyū-buntansha) |
KUBOTA Shunichiro University of Tokyo, Graduate School of Medicine, Associate Professor, 大学院・医学系研究科, 助教授 (00260480)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥15,000,000 (Direct Cost: ¥15,000,000)
Fiscal Year 2000: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 1999: ¥11,500,000 (Direct Cost: ¥11,500,000)
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| Keywords | Cholestanol / Apoptosis / Cerebellar neuronal cell / Cerebrotendinous xanthomatosis / cornea endothelial cells / lens epithelial cells / cerebellar ataxia / cataract / CTX / プルキンエ細胞 / インターロイキン1β / CPP32プロテアーゼ / 小脳症状 / インターロイキン 1β / CPP32 プロテアーゼ |
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| Research Abstract |
Cerebrotendinous xanthomatosis (CTX) is a hereditary lipid storage disease characterized by hyper-cholestanolemia, crebellar ataxia, xanthoma, and cataract. We hypothesized that cholestanol in serum of CTX patients might induce neuronal cell death in cerebellum and eventually lead to cerebellar ataxia. To elucidate this hypothesis we have established hyper-cholestanolemia rats by feeding cholestanol. Neuronal cells, especially Purkinje cells in the cerebellum were stained by Sudan black B only in cholestanol-fed rats, indicating the deposit of cholestanol in cerebellum. To examine effects of cholestanol in vitro, cerebellar neuronal cells were cultured with cholestanol. The cholestanol concentration increased and the viability decreased in cells cultured with cholestanol. Apoptosis was detected in cells cultured with cholestanol more frequently than in control cells using terminal deoxynucleotidyl transferase (TdT) dUTP nick end-labeling (TUNEL) method. The activities of interleukin-1β
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-converting enzyme (ICE) and CPP32 protease were increased in cells cultured with cholestanol. We further studied the effect of cholestanol on induce cornea endothelial and lens epithelial cell death in vitro. Both cornea endothelial and lens epithelial cells cultured with 10 μg/ml cholestanol showed a significant loss of viability. The nuclei of these cells cultured with 10 μg/ml cholestanol were more frequently stained than those exposed to 10 u」/ml cholesterol or 1 % ethanol. Quantitative analysis of apoptotic DNA fragmentation confirmed that the cholestanol induced apoptosis of these cells in a time-dependent manner. The activities of ICE and CPP32 proteases for cells cultured with 10 μg/ml cholestanol were significantly higher than those observed in control cells. In vitro, cholestanol was taken up by cornea endothelial cells and lens epithelial cells, an event which led to apoptosis of these cells.
The data suggest that cholestanol induced apoptosis of cerebellar neuronal cells, cornea endothelial cells and lens epithelial cells. The results in the present study may explain the mechanism of cerebellar ataxia and cataract of CTX patients. Less
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