TRAF6沉默对LPS刺激牙周膜成纤维细胞增殖能力的影响

发布时间：2018-04-09 03:31

本文选题：肿瘤坏死因子受体相关因子　切入点：RNA干扰　出处：《口腔医学研究》2017年04期

【摘要】：目的:使用小干扰RNA(small interfering RNA,siRNA)沉默人牙周膜成纤维细胞(human periodontal ligament cell,hPDLC)肿瘤坏死因子受体相关因子6(tumor necrosis factor receptor associated factor 6,TRAF6)的基因,观察细胞的增殖情况及脂多糖刺激时增殖能力的变化。方法:酶消化联合组织块法体外培养原代hPDLC,并进行免疫组化鉴定,将细胞分为TRAF6siRNA组、control siRNA组、NONE组,脂质体法将TRAF6siRNA、control siRNA分别转染入对应组中,NONE组只加入等量的转染试剂,再使用10mg/L LPS刺激细胞,RT-PCR检测TRAF6mRNA的表达情况,CCK-8检测沉默前后及LPS刺激时细胞增殖能力的改变。结果:免疫组化鉴定hPDLC培养成功;TRAF6siRNA组的TRAF6mRNA的表达水平与NONE、control siRNA组相比显著下降(P0.001);TRAF6siRNA组在转染后36、48、72h的A值均显著低于相同时间点的2个对照组(P0.001);使用10mg/L LPS刺激各组细胞24、48h时,TRAF6siRNA组细胞的A值显著低于相同时间点的2个对照组(P0.001)。结论:沉默TRAF6基因能抑制LPS刺激时hPDLC的增殖,推测TRAF6可能影响牙周炎的发生发展进程,是牙周炎潜在的治疗靶点。
[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.
【作者单位】：山西医科大学口腔医学系;
【基金】：山西省科技攻关项目(编号:20150313010-3) 山西医科大学校科技创新基金项目(编号:02101313);山西医科大学博士启动基金项目(编号:03201321)
【分类号】：R781.4

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