TRAF6沉默对LPS刺激牙周膜成纤维细胞成骨分化的影响

doi:10.13701/j.cnki.kqyxyj.2017.11.012

口腔医学研究 ›› 2017, Vol. 33 ›› Issue (11): 1181-1184.DOI: 10.13701/j.cnki.kqyxyj.2017.11.012

TRAF6沉默对LPS刺激牙周膜成纤维细胞成骨分化的影响

陈庆勇, 李霞^*, 张芳, 王锦华

山西医科大学口腔医学系山西太原 030001

收稿日期:2017-05-07 出版日期:2017-11-20 发布日期:2017-11-29
通讯作者: 李霞,E-mail:lixia6881@163.com
作者简介:陈庆勇(1988~ ),男,山东聊城人,硕士,医师,主要从事牙周病的基础研究。
基金资助:
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)

Effect of TRAF6 Knockdown on Osteogenic Differentiation of Human Periodontal Ligament Cell Treated with LPS

CHEN Qing-yong, LI Xia^*, ZHANG Fang, WANG Jin-hua

Department of Stomatology, Shanxi Medical University, Taiyuan 030001, China

Received:2017-05-07 Online:2017-11-20 Published:2017-11-29

摘要/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。

Abstract: Objective: To observe the effect of tumor necrosis factor receptor-related factor 6 (TRAF6) knockdown on osteogenic differentiation of human periodontal ligament cell (hPDLC) stimulated by LPS. Methods: The TRAF6 siRNA and control siRNA were transiently transfected into hPDLC. The cells were stimulated with 10μg/ml LPS. The silence efficiency of TRAF6 was detected by RT-PCR and Western blot. The expression of ALP was detected by alkaline phosphatase assay kit. The expression of Runx-2 and type I collagen (Col-I) gene was detected by RT-PCR. Results: The expression of TRAF6 mRNA and protein in TRAF6 siRNA group was significantly decreased (P<0.001). The expression of ALP in LPS-stimulated group was significantly lower than that in non-LPS-stimulated group (P<0.05). The expression of ALP, Runx-2, and Col-I mRNA in TRAF6 siRNA was significantly higher than that in the other three groups (P<0.05). Conclusion: TRAF6 knockdown can alleviate the inhibitory effect of LPS on osteogenesis differentiation of hPDLC, which indicates that TRAF6 may affect the development of periodontitis and may be a potential therapeutic target for periodontitis.

Key words: Tumor necrosis factor, Receptor associated factor 6, RNA interfering, Osteogenic differentiation, Lipopolysaccharides

中图分类号:

R781.4

陈庆勇, 李霞, 张芳, 王锦华. TRAF6沉默对LPS刺激牙周膜成纤维细胞成骨分化的影响[J]. 口腔医学研究, 2017, 33(11): 1181-1184.

CHEN Qing-yong, LI Xia, ZHANG Fang, WANG Jin-hua. Effect of TRAF6 Knockdown on Osteogenic Differentiation of Human Periodontal Ligament Cell Treated with LPS[J]. Journal of Oral Science Research, 2017, 33(11): 1181-1184.

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TRAF6沉默对LPS刺激牙周膜成纤维细胞成骨分化的影响

Effect of TRAF6 Knockdown on Osteogenic Differentiation of Human Periodontal Ligament Cell Treated with LPS

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