2019 Fiscal Year Final Research Report
Development of a novel cholesterol efflux assay method as neither cell culture, radioisotope nor ultracentrifugation is required
| Project/Area Number |
17K08975
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Laboratory medicine
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| Research Institution | Shinshu University (2019)
Tokyo Medical and Dental University (2017-2018) |
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Principal Investigator |
Tozuka Minoru 信州大学, 医学部, 特任教授 (60431954)
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| Co-Investigator(Kenkyū-buntansha) |
大川 龍之介 東京医科歯科大学, 大学院医歯学総合研究科, 教授 (50420203)
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| Project Period (FY) |
2017-04-01 – 2020-03-31
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| Keywords | cholesterol efflux / high-density lipoprotein / apoB-depleted serum / gel beads / cardiovascular disease |
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| Outline of Final Research Achievements |
Cholesterol efflux capacity (CEC) is the key metric for determining the antiatherosclerotic function of high-density lipoprotein (HDL). However, the assay methods currently used to calculate CEC are not ideal for clinical use as they require the culture of cells.In the present study, we developed novel CEC assay using immobilized liposome-bound gel beads (ILG), containing fluorescently labeled cholesterol, as a substitute for cultured cells. The CEC values obtained with this ILG method were correlated with those obtained with a conventional method using THP-1 macrophages derived foam cells and radioisotope labeled-cholesterol as a tracer. Overall, this novel CEC assay method is a realistic and effective alternative to current methods in the field while also being easier to use in clinical laboratories as neither cell culture, radioisotope nor ultracentrifugation is required.
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| Free Research Field |
臨床化学
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| Academic Significance and Societal Importance of the Research Achievements |
近年、HDL量の指標となるHDL-コレステロール検査が必ずしも抗粥状動脈硬化の指標にならず、むしろその機能が重要であると考えられている。HDLの代表的な抗粥状動脈硬化機能は、泡沫細胞に過剰蓄積されたコレステロールを肝臓に運ぶ、いわゆるコレステロール逆輸送である。この第１段階が細胞からのコレステロール引き抜き能（CEC）である。現在のCEC測定法は臨床検査室で日常検査として用いるには、培養細胞を用いるなどの制約から実施が困難である。本研究では培養細胞に代わる蛍光標識コレステロールを含むゲルビーズを開発した。この方法は臨床検査室でHDLの機能を測定する簡易法開発の足がかりになることが期待される。
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