タバコ煙刺激と歯周病原細菌によるヒト歯肉上皮細胞のmiRNA発現・機能解析

香烟烟雾刺激和牙周病原菌诱导的人牙龈上皮细胞miRNA表达及功能分析

• 批准号: 15H06631
• 负责人: 今村 健太郎
• 金额: $ 1.75万
• 依托单位: Tokyo Dental College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Research Activity Start-up
• 财政年份: 2015
• 资助国家: 日本
• 起止时间: 2015-08-28 至 2017-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15H06631/
• 关键词: miRNA; タバコ煙; Porphyromonas gingivalis; ヒト歯肉上皮細胞; 細胞遊走能

我々はこれまでタバコ煙抽出物（CSC）が細胞機能，とくに遊走能に対し低濃度で促進，高濃度で抑制と二相性の影響を与え，このようなCSC の作用はPorphyromonas gingivalis感染によって修飾されることを明らかにしてきた。この先行研究の結果を受け，さらに詳細なメカニズムを検討することを目的として，本年度は宿主細胞に対するCSCと P. gingivalis 感染によるエピジェネッティックな影響を検索した。遊走能の促進が認められた低濃度CSC10 μg/ml によるヒト歯肉上皮細胞のmiRNA 発現の変化をPCR arrayを用いて評価した。その結果を踏まえ，変化が著名に認められたmiRNAの中から，細胞遊走の促進と抑制への関与が示唆されているものを選択し，高濃度CSC250 μg/ml とP. gingivalis 感染を伴わせた条件を加えてRT-PCR を行った。PCR arrayの結果として，84個のmiRNAうち64個において1.5倍以上の上昇が認められた。さらにRT-PCRでは，遊走能の促進に関与しているmiR-125aの発現量をコントロールと比較したところ，低濃度CSC群で1.4倍，高濃度CSC＋P. gingivalis 群で1.7倍の発現上昇を示し，高濃度CSCでは60％程度の減少を認めた。また，抑制に関与が示唆されているmiR-301は高濃度CSC＋P. gingivalis 群において1.9倍の発現量を示した。以上の結果より，miR-125aは低濃度CSCによる細胞遊走能の抑制に関与しており，miR-301は高濃度CSCとP. gingivalis 感染による遊走能の抑制に関与していることが明らかとなった。今後は，これらのmiRNAがシグナル伝達経路（MAPkinase）関連タンパクに与える影響を解明し，宿主細胞の機能，特に細胞増殖能や遊走能の制御法の検討をする必要があると考えている。

We use the extractive (CSC) to measure the mechanical properties of the cells, the walking energy of the vehicle, the promotion of low temperature, the inhibition of dimorphism at high temperature, the effect of CSC on Porphyromonas gingivalis infection, and the effect of Porphyromonas gingivalis infection. The results of the first study were successful. This year, the host cell was infected with CSC P. the results of the study were successful. This year, the host cell is infected with P. gingivalis infection. The walking energy promotes the development of low-temperature CSC10 μ g/ml cells in the meat epithelium and miRNA in the meat epithelium to induce the transformation of PCR array. The results showed that the well-known drugs in miRNA were optimized, the inhibition and instigation were promoted by cytopathic acid, and the infection with high degree of CSC250 μ g/ml P. gingivalis infection was accompanied by the addition of RT-PCR conditions. The results of PCR array results were successful, and 84 miRNA two-tier clusters were more than 1.5 times higher than that of the previous one. The speed of RT-PCR, walking energy can promote the performance of low-level CSC, high-degree CSC+P. Gingivalis group, low-degree CSC group, high-degree CSC+P group, low-degree gingivalis group, low-degree QR group, high-degree QR group, low-degree QR group, high-level QR group, low-degree QR group, high-degree QR group, low-degree QR group, high-level QR group, high-level QR group, high-degree QR group, low-degree QR group, high-degree CSC+P group, Check, suppress and instigate the high degree of CSC+P of miR-301. Gingivalis group is 1.9times higher than that of the control group. The results of the above results showed that the cellular walking ability of miR-125a with low concentration of CSC was inhibited, while that of miR-301 with high intensity of CSC, P. gingivalis infection was significantly decreased. In the future, you will need to know that the host cell has the ability to do so. In the future, you will need to know that the host cell has the ability to do so. In the future, you will need to know that the host cell has the ability to do so. In the future, you will be able to determine the ability of the host cell.