JP7397541B1 - Nadh-ユビキノン酸化還元酵素サブユニット6遺伝子発現亢進用医薬組成物及び食品組成物 - Google Patents
Nadh-ユビキノン酸化還元酵素サブユニット6遺伝子発現亢進用医薬組成物及び食品組成物 Download PDFInfo
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Abstract
Description
本試験では、ヒト肝臓由来細胞 Hep G2を5-アセチル-1-ベンジルピロリジン-2-オン存在下で培養した後 、DNAマイクロアレイ解析を実施し、ND6遺伝子発現亢進作用があるか否かを確認した。
ヒト肝癌株 Hep G2
国立研究開発法人 医薬基盤・健康・栄養研究所より入手。
1)5-アセチル-1-ベンジルピロリジン-2-オン10 mg にDMSO を100 μL 加え溶解し,DMSO で被験物質の濃度が50 mMとなるよう段階希釈した。
2)さらに細胞培養用培地で被験物質の濃度が50 μMとなるよう希釈した。
〈Hep G2細胞の調製〉
1)Hep G2 細胞を2×105 cell/mL の細胞濃度で6 cm dish に5 mL ずつ播種した。
2)播種24 時間後に接着を確認し、細胞培養用培地を除去後、新たに被験物質(最終濃度50 μM)含有の細胞培養用培地を各5 mL ずつ添加した。
3)添加後24 時間培養し、細胞培養用培地を抜いて接着細胞をPBS で洗浄後、得られた細胞を下述のRNA 抽出→DNA マイクロアレイ解析に用いた。
1)上述の細胞調製で得られた細胞から、RNAiso plus を用いてRNA を抽出し、RNeasy MinElute Cleanup Kitで精製した。
1)RNA サンプルをLow Input Quick Amp Labeling Kit (Agilent) を用いてcDNA の合成、Cy3 ラベル化cRNA 合成と精製を行った。
2)Gene Expression Hybridization Kit (Agilent) を用い、それぞれのラベル化cRNA をフラグメンテーションし,Whole Human Genome Microarray Ver2.0 (Agilent) にアプライ、65℃で17 時間ハイブリダイゼーションした。
3)Gene Expression Wash Buffer 1 及び2 (Agilent) を用い、アレイスライドを洗浄した。
4)マイクロアレイスキャナー (GenePix 4000B,Molecular Devices) でスキャンしたアレイ画像を、アレイ解析ソフトウェアGenePix Pro (Molecular Devices) で数値化した。蛍光強度値をノーマライズし、コントロールに対して被験物質添加群のRatioを算出した。
ND6遺伝子はヒト肝臓由来細胞株(HepG2)において5-アセチル-1-ベンジルピロリジン-2-オン共存下では発現が1.66倍亢進した。よって、5-アセチル-1-ベンジルピロリジン-2-オンはND6遺伝子発現を亢進することでND6の産生を促進すると考えられる。
Claims (2)
- 5-アセチル-1-ベンジルピロリジン-2-オンから成るNADH-ユビキノン酸化還元酵素サブユニット6遺伝子発現亢進用医薬組成物。
- 5-アセチル-1-ベンジルピロリジン-2-オンから成るNADH-ユビキノン酸化還元酵素サブユニット6遺伝子発現亢進用食品組成物。
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JP7134541B1 (ja) * | 2022-05-30 | 2022-09-12 | Towa Corporation 株式会社 | キサンチンオキシダーゼ阻害用医薬組成物及び食品組成物 |
JP2022184813A (ja) * | 2021-05-31 | 2022-12-13 | 高雄醫學大學 | マカ抽出物及びその用途 |
JP7253301B1 (ja) * | 2023-01-27 | 2023-04-06 | Towa Corporation 株式会社 | 5-アミノレブリン酸産生促進用医薬組成物及び食品組成物 |
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Decoding MultipleBiofunctions of Maca on Its Anti-allergic, Anti-inflammatory, Anti-thrombotic,and Pro-angiogenic Activities,Journal of Agricultural and Food Chemistry,2021年,69(40),11856-11866 |
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