JP7371996B1 - インスリン様成長因子2遺伝子発現亢進用医薬組成物及び食品組成物 - Google Patents
インスリン様成長因子2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、ヒト副腎皮質由来細胞 H295R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、IGF2遺伝子発現亢進作用があるか否かを確認した。
ヒト副腎皮質腺がん株(H295R)
American Type Culture Collection(ATCC)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。
〈H295R細胞の調製〉
1)H295R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。
マイクロアレイを活用した網羅的遺伝子発現解析の結果、IGF2遺伝子は副腎皮質由来細胞(H295R)において5-アセチル-1-ベンジルピロリジン-2-オン共存下で発現が1.59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IGF2遺伝子発現を亢進することでIGF2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-オンを有効成分として含有してなるインスリン様成長因子2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-オンを有効成分として含有してなるインスリン様成長因子2遺伝子発現亢進用食品組成物。
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Citations (4)
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JP6537689B1 (ja) * | 2018-10-26 | 2019-07-03 | Towa Corporation 株式会社 | エストラジオール産生促進用医薬組成物及び食品組成物 |
JP6578046B1 (ja) * | 2018-08-23 | 2019-09-18 | Towa Corporation 株式会社 | Pde5阻害用医薬組成物及びpde5阻害用食品組成物 |
JP6962630B1 (ja) * | 2021-04-01 | 2021-11-05 | Towa Corporation 株式会社 | 筋肥大促進用医薬組成物及び食品組成物 |
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