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- 2017
TRAF6沉默对LPS刺激牙周膜成纤维细胞成骨分化的影响
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Abstract:
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,观察沉默TRAF6基因对LPS刺激的hPDLC成骨分化的影响。方法:实验分为空白对照组、转染试剂组、TRAF6 siRNA组、control siRNA组,采用脂质体法将TRAF6 siRNA、control siRNA分别瞬时转染入对应组中,设转染试剂组加入等量的转染试剂,空白对照组不做处理,再使用10 mg/L LPS刺激细胞,RT-PCR、Western blot检测转染后TRAF6的基因及蛋白表达水平,使用LPS+成骨诱导液培养细胞,碱性磷酸酶检测试剂盒检测碱性磷酸酶的活性,RT-PCR检测Runx-2和I型胶原蛋白(Col-I)基因表达情况。结果:TRAF6 siRNA组的TRAF6mRNA和蛋白的表达显著降低(P<0.001);成骨诱导3 d后,LPS刺激下各组ALP的表达量要显著低于对照组(P<0.05),TRAF6 siRNA组的ALP表达量要高于空白对照、转染试剂组和control siRNA组(P<0.05),TRAF6 siRNA的Runx-2和Col-I的mRNA表达均明显高于其余3组(P<0.05)。 结论:沉默TRAF6基因能减轻LPS对hPDLC成骨分化的抑制作用,即沉默TRAF6基因能促进LPS刺激下的hPDLC成骨分化,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
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