miR-495-3p对LPS刺激人牙周膜细胞增殖及炎性因子表达的影响

›› 2017, Vol. 8 ›› Issue (2): 86-89.

miR-495-3p对LPS刺激人牙周膜细胞增殖及炎性因子表达的影响

张帆¹,余国玺²,王力锋³

1. 西北工业大学医院
2. 西北工业大学医院口腔科
3. 西安小白兔口腔医院种植科

收稿日期:2017-03-31 修回日期:2017-06-07 出版日期:2017-06-25 发布日期:2017-07-03
通讯作者: 张帆 E-mail:fan_zhang029@qq.com

Effect of miR-495-3p on the proliferation of hPDLC and the production of inflammatory cytokines induced by LPS

Received:2017-03-31 Revised:2017-06-07 Online:2017-06-25 Published:2017-07-03

摘要/Abstract

摘要： 目的：研究miR-495-3p对脂多糖（LPS）刺激人牙周膜细胞增殖及炎性因子的影响。方法：分离培养人原代牙周膜细胞（hPDLC），然后给予10 μg/mL的LPS处理及miR-495-3p模拟物（mimic）转染。实时定量RT-PCR检测LPS处理后miR-495-3p及Toll样受体4（TLR4）的表达；MTT法检测hPDLC细胞增殖；ELISA法检测白细胞介素6（IL-6）及肿瘤坏死因子α（TNF-α）的含量；双荧光素酶报告基因检测miR-495-3p与TLR4的靶向关系；Western blot法检测miR-495-3p mimic转染后TLR4及磷酸化核因子κB抑制蛋白α（p-IκBα）的表达。结果：LPS处理后，miR-495-3p的表达显著下降，TLR4的表达显著上升。过表达miR-495-3p可显著促进hPDLC细胞的增殖，抑制IL-6及TNF-α的产生；miR-495-3p可靶向结合到TLR4的3' UTR区，并下调TLR4的表达；过表达miR-495-3p可显著抑制p-IκBα的表达。结论：miR-495-3p可通过靶向下调TLR4的表达，抑制核转录因子-κB（NF-κB）通路，进而抑制炎性因子的产生，促进hPDLC的增殖。

Abstract: Objective：To analyze the effect of miR-495-3p on the proliferation of hPDLC and the production of inflammatory cytokines that induced by LPS. Methods：The primary hPDLC was isolated and was treated with LPS or LPS+miR-495-3p mimic transfection. The of miR-495-3p and TLR4 was measured by Real-time PCR. The proliferation of hPDLC was measured by MTT method. The production of IL-6 and TNF-α was measured by ELISA. The target relationship between miR-495-3p and TLR4 was measured by dual luciferase assay. The of TLR4 and IκBa was measured by Western blot. Results: LPS significantly down-regulated the of miR-495-3p and up-regulated the of TLR4. MiR-495-3p mimic dramatically increased the proliferation of hPDLC, decreased the production of IL-6 and TNF-α. MiR-495-3p targeted the 3' UTR region of TLR4. MiR-495-3p mimic significantly decreased the of TLR4. Furthermore, miR-495-3p mimic noticeably suppressed the of p-IκBα. Conclusions：miR-495-3p increased the proliferation of hPDLC and suppressed the production of inflammatory cytokines through down regulated the of TLR4 and inhibited NF-κB pathway.

张帆余国玺王力锋. miR-495-3p对LPS刺激人牙周膜细胞增殖及炎性因子表达的影响[J]. , 2017, 8(2): 86-89.

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