本文選題：腫瘤壞死因子受體相關(guān)因子　切入點：RNA干擾　出處：《口腔醫(yī)學(xué)研究》2017年04期

【摘要】：目的:使用小干擾RNA(small interfering RNA,siRNA)沉默人牙周膜成纖維細(xì)胞(human periodontal ligament cell,hPDLC)腫瘤壞死因子受體相關(guān)因子6(tumor necrosis factor receptor associated factor 6,TRAF6)的基因,觀察細(xì)胞的增殖情況及脂多糖刺激時增殖能力的變化。方法:酶消化聯(lián)合組織塊法體外培養(yǎng)原代hPDLC,并進(jìn)行免疫組化鑒定,將細(xì)胞分為TRAF6siRNA組、control siRNA組、NONE組,脂質(zhì)體法將TRAF6siRNA、control siRNA分別轉(zhuǎn)染入對應(yīng)組中,NONE組只加入等量的轉(zhuǎn)染試劑,再使用10mg/L LPS刺激細(xì)胞,RT-PCR檢測TRAF6mRNA的表達(dá)情況,CCK-8檢測沉默前后及LPS刺激時細(xì)胞增殖能力的改變。結(jié)果:免疫組化鑒定hPDLC培養(yǎng)成功;TRAF6siRNA組的TRAF6mRNA的表達(dá)水平與NONE、control siRNA組相比顯著下降(P0.001);TRAF6siRNA組在轉(zhuǎn)染后36、48、72h的A值均顯著低于相同時間點的2個對照組(P0.001);使用10mg/L LPS刺激各組細(xì)胞24、48h時,TRAF6siRNA組細(xì)胞的A值顯著低于相同時間點的2個對照組(P0.001)。結(jié)論:沉默TRAF6基因能抑制LPS刺激時hPDLC的增殖,推測TRAF6可能影響牙周炎的發(fā)生發(fā)展進(jìn)程,是牙周炎潛在的治療靶點。
[Abstract]:Aim: to silence the tumor necrosis factor receptor related factor 6(tumor necrosis factor receptor associated factor 6 (TRAF6) gene in human periodontal ligament fibroblasts (HPDLC) by using small interfering RNA(small interfering siRNAs, and to observe the proliferation of human periodontal ligament fibroblasts and the changes of lipopolysaccharide stimulated proliferation.Methods: the primary hPDLC cells were cultured in vitro by enzyme digestion combined with tissue block method and identified by immunohistochemistry. The cells were divided into control siRNA group and none group. The TRAF6 siRNA control siRNA was transfected into the corresponding group by liposome method, and only the same amount of transfection reagent was added to the control siRNA group by liposome method.Then the expression of TRAF6mRNA was detected by 10mg/L LPS stimulated cell RT-PCR and the change of cell proliferation ability before and after LPS stimulation was detected by CCK-8.Results: compared with the control siRNA group, the level of TRAF6mRNA expression in the hPDLC cultured successfully TRAF6 siRNA group was significantly lower than that in the control siRNA group. The A value of the TRAF6 siRNA group after transfection was significantly lower than that in the control group at the same time point (P 0.001), and 10mg/L LPS was used to stimulate the cells in each group.The A value of TRAF6 siRNA group was significantly lower than that of two control groups at 24 h.Conclusion: silencing TRAF6 gene can inhibit the proliferation of hPDLC stimulated by LPS. It is speculated that TRAF6 may influence the development of periodontitis and may be a potential therapeutic target for periodontitis.
【作者單位】： 山西醫(yī)科大學(xué)口腔醫(yī)學(xué)系;
【基金】：山西省科技攻關(guān)項目(編號:20150313010-3) 山西醫(yī)科大學(xué)�？萍紕�(chuàng)新基金項目(編號:02101313);山西醫(yī)科大學(xué)博士啟動基金項目(編號:03201321)
【分類號】：R781.4

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