【摘要】：目的:研究雷帕霉素對(duì)人牙周膜干細(xì)胞中miR-30a的表達(dá)的影響,并驗(yàn)證其與Beclin1的靶向調(diào)控關(guān)系,探討miR-30a與自噬途徑在牙周膜干細(xì)胞的炎癥作用機(jī)制。方法:分離培養(yǎng)人牙周膜干細(xì)胞,并分別進(jìn)行50μg/L雷帕霉素作用2h,10nmol/L 3-甲基腺嘌呤(3-MA)作用12h,收集細(xì)胞,提取總RNA;應(yīng)用實(shí)時(shí)熒光定量PCR(qRT-PCR)檢測(cè)miR-30a的表達(dá)水平;應(yīng)用qRT-PCR、Western blot及雙熒光素酶報(bào)告系統(tǒng)驗(yàn)證miR-30a與Beclin1的相互作用關(guān)系。結(jié)果:雷帕霉素刺激細(xì)胞后,細(xì)胞內(nèi)的miR-30a的表達(dá)升高;qRT-PCR、Western blot及雙熒光素酶報(bào)告系統(tǒng)證實(shí),miR-30a對(duì)Beclin1的mRNA及蛋白均有抑制,且與Beclin1的3’UTR具有靶向抑制作用。結(jié)論:miR-30a可靶向抑制自噬相關(guān)基因Beclin1的表達(dá),并參與人牙周膜干細(xì)胞的細(xì)胞自噬過程。
[Abstract]:Aim: to investigate the effect of rapamycin on the expression of miR-30a in human periodontal ligament stem cells (PDSCs), and to verify the relationship between rapamycin and Beclin1, and to explore the inflammatory mechanism of miR-30a and autophagy pathway in periodontal ligament stem cells (PDSCs). Methods: human periodontal ligament stem cells were isolated and cultured, and treated with 50 渭 g / L rapamycin for 2 h or 10 nmol / L 3-methyladenine (3-MA) for 12 h. The total RNA; was collected and the expression of miR-30a was detected by real-time fluorescence quantitative PCR (qRT-PCR). QRT-PCR,Western blot and double luciferase report system were used to verify the interaction between miR-30a and Beclin1. Results: after stimulated by rapamycin, the expression of miR-30a in the cells was increased. The expression of QRT-PCR blot and double luciferase report system confirmed that pmiR-30a could inhibit the mRNA and protein of Beclin1, and had a targeted inhibitory effect with 3'UTR of Beclin1. Conclusion: 1 miR-30a can inhibit the expression of autophagy related gene Beclin1 and participate in the autophagy process of periodontal ligament stem cells.
【作者單位】： 首都醫(yī)科大學(xué)附屬北京康復(fù)醫(yī)院口腔科;北京大學(xué)口腔醫(yī)院綜合科;
【分類號(hào)】：R781.4

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