TRAF6沉默对LPS刺激牙周膜成纤维细胞增殖能力的影响

doi:10.13701/j.cnki.kqyxyj.2017.04.003

口腔医学研究 ›› 2017, Vol. 33 ›› Issue (4): 358-361.DOI: 10.13701/j.cnki.kqyxyj.2017.04.003

TRAF6沉默对LPS刺激牙周膜成纤维细胞增殖能力的影响

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:2016-09-26 出版日期:2017-04-20 发布日期:2017-04-24
通讯作者: 李霞,E-mail:lixia6881@163.com
作者简介:陈庆勇(1988~ ),男,山东省聊城市人,硕士在读,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 Silencing by siRNA on Proliferation of Human Periodontal Ligament Cells Treated with LPS.

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

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

Received:2016-09-26 Online:2017-04-20 Published:2017-04-24

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TRAF6 siRNA组、control siRNA组、NONE组,脂质体法将TRAF6 siRNA、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10 mg/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;TRAF6 siRNA组的TRAF6 mRNA的表达水平与NONE、control siRNA组相比显著下降(P<0.001);TRAF6 siRNA组在转染后36、48、72 h的A值均显著低于相同时间点的2个对照组(P<0.001);使用10 mg/L LPS刺激各组细胞24、48 h时,TRAF6 siRNA组细胞的A值显著低于相同时间点的2个对照组(P<0.001)。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: Objective: To investigate the effect of siRNAtargeting TRAF6 on proliferation of human periodontal ligament cells(hPDLC) under LPS challenge. Methods: hPDLC was isolated from human periodontal ligament. The immunohistochemistry was used to examinethe expression of cytokeratin and vimentin. Lipofectamine was used to transfect TRAF6 siRNA and control siRNA into hPDLC, and then the cells were stimulated by LPS. RT-PCR was employed to determine TRAF6 mRNA. CCK-8 was used to test the proliferation of hPDLC under LPS challenge. Results: Immunohistochemistry confirmed that the cells were hPDLC. Compared with the control group, the mRNA expression of TRAF6 in TRAF6 siRNA group was decreased. The A value of TRAF6 knockdown cells was significantly decreased compared with thecontrol group(P<0.001).The proliferation of TRAF6 siRNA group treated with 10μg /mL LPS was also significantly decreased compared with other groups(P<0.001). Conclusion: The results suggest that TRAF6 knockdown inhibits the proliferation upon LPS challenge, which indicates thatTRAF6 maybe a target for periodontitis therapy.

Key words: TRAF6 , siRNA , Lipopolysaccharides, Cell proliferation

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增殖能力的影响[J]. 口腔医学研究, 2017, 33(4): 358-361.

CHEN Qing-yong, LI Xia, ZHANG Fang,WANG Jin-hua.. Effect of TRAF6 Silencing by siRNA on Proliferation of Human Periodontal Ligament Cells Treated with LPS.[J]. Journal of Oral Science Research, 2017, 33(4): 358-361.

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TRAF6沉默对LPS刺激牙周膜成纤维细胞增殖能力的影响

Effect of TRAF6 Silencing by siRNA on Proliferation of Human Periodontal Ligament Cells Treated with LPS.

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