本文選題：尼古丁 + 牙周膜干細胞�。� 參考：《華西口腔醫(yī)學雜志》2017年04期

【摘要】：目的探討尼古丁對牙周膜干細胞(PDLSCs)增殖和成骨分化能力的調(diào)控作用,檢測尼古丁能否通過調(diào)控Toll樣受體4(TLR4)抑制PDLSCs的增殖及成骨分化能力。方法培養(yǎng)PDLSCs,采用流式細胞儀檢測PDLSCs表面抗原標志,WST-1試劑盒檢測不同濃度尼古丁刺激后PDLSCs增殖能力的改變。采用茜素紅染色觀察PDLSCs經(jīng)不同濃度尼古丁刺激和成骨誘導后的礦化結(jié)節(jié)生產(chǎn)情況。運用實時定量聚合酶鏈反應(RT-PCR)和Western blot檢測經(jīng)尼古丁刺激后,PDLSCs成骨能力相關(guān)基因和蛋白的改變,以及尼古丁聯(lián)合TLR4抑制劑TAK-242刺激后,PDLSCs成骨能力相關(guān)基因及蛋白的改變。結(jié)果培養(yǎng)的細胞表達間充質(zhì)干細胞表面標志物CD90和CD105,證實為PDLSCs。與對照組相比,培養(yǎng)3 d后,尼古丁濃度為10-4 mol·L-1的PDLSCs的增殖能力受到明顯抑制(P0.05);成骨誘導21 d后,10-4 mol·L-1尼古丁刺激組茜素紅染色礦化結(jié)節(jié)明顯減少。RT-PCR反應及Western blot顯示:與對照組相比,尼古丁刺激組PDLSCs的堿性磷酸酶、骨鈣素、Runt相關(guān)轉(zhuǎn)錄因子2的基因及蛋白表達量均降低(P0.05),加入TLR4抑制劑TAK-242后,尼古丁的抑制效應減弱。結(jié)論尼古丁可能通過TLR4信號通路抑制PDLSCs的增殖及成骨分化能力。
[Abstract]:Objective to investigate the effects of nicotine on the proliferation and osteogenic differentiation of periodontal ligament stem cells (PDLSCs), and to investigate whether nicotine can inhibit the proliferation and osteogenic differentiation of PDLSCs by regulating Toll-like receptor 4 (TLR4). Methods PDLSCs were cultured and the proliferation of PDLSCs was detected by flow cytometry. The mineralized nodules of PDLSCs stimulated by different concentrations of nicotine and induced by osteogenesis were observed by alizarin red staining. Real-time quantitative polymerase chain reaction (RT-PCR) and Western blot were used to detect the changes of osteoblast related genes and proteins in PDLSCs stimulated by nicotine and TLR4 inhibitor TAK-242. Results the cultured cells expressed CD90 and CD105, which were confirmed as PDLSCs. Compared with the control group, the cells were cultured for 3 days. The proliferation of PDLSCs with 10 ~ (-4) mol L ~ (-1) of nicotine was significantly inhibited (P0.05), and the mineralized nodules of alizarin red staining were significantly decreased in the group stimulated by nicotine at 10 ~ (-4) mol L ~ (-1) after osteogenesis (P0.05). The results of RT-PCR and Western blot showed that: compared with the control group, there was no significant difference between the two groups. The expression of alkaline phosphatase and osteocalcin Runt related transcription factor 2 gene and protein in PDLSCs were decreased in nicotine stimulation group (P0.05). The inhibitory effect of nicotine on PDLSCs was weakened by adding TLR4 inhibitor TAK-242. Conclusion nicotine may inhibit the proliferation and osteogenic differentiation of PDLSCs through TLR4 signaling pathway.
【作者單位】： 重慶醫(yī)科大學附屬口腔醫(yī)院牙體牙髓病科口腔疾病與生物醫(yī)學重慶市重點實驗室重慶市高校市級口腔生物醫(yī)學工程重點實驗室;
【基金】：國家自然科學基金面上項目(31571508,31371473) 重慶市第七批重點學科建設(shè)項目(牙體牙髓病學,2011年) 重慶市衛(wèi)生局醫(yī)學科研計劃重點項目(2011-1-062) 重慶市渝北區(qū)科技項目[渝北財教(2011)33號]~~
【分類號】：R781.4

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