NF-κB-Regulated miR-99a Modulates Endothelial Cell Inflammation.
NF-κB-Regulated miR-99a Modulates Endothelial Cell Inflammation.
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DOI：
10.1155/2016/5308170
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发表时间：
2016
影响因子：
4.6
通讯作者：
Zhou HH
中科院分区：
文献类型：
--
作者：
Bao MH;Li JM;Luo HQ;Tang L;Lv QL;Li GY;Zhou HH
目标。本研究旨在探讨miR-99a在lps诱导的内皮细胞炎症中的作用和机制，以及NF-κB对miR-99a产生的调控作用。方法与结果。ELISA结果显示，LPS处理显著促进炎症因子（TNF-α、IL-6、IL-1β和MCP-1）的分泌。LPS处理也抑制了miR-99a的产生，促进了mTOR的表达和NF-κB核易位。过表达miR-99a可抑制lps诱导的TNF-α、IL-6、IL-1β和MCP-1过量产生、mTOR上调和NF-κB核易位。PROMO软件分析显示，NF-κB结合位点位于miR-99a启动子的−1643 ~−1652区域。双荧光素酶报告基因分析、电泳迁移率转移实验（EMSA）和染色体免疫沉淀（ChIP）实验表明，NF-κB通过结合miR-99a启动子的- 1643至- 1652区域促进miR-99a的转录。对HUVECs的进一步研究证实了NF-κB对miR-99a产生的调控作用。结论。MiR-99a通过抑制mTOR/NF-κB信号抑制lps诱导的HUVECs炎症。NF-κB通过结合miR-99a启动子的- 1643至- 1652区域促进miR-99a的产生。考虑到内皮炎症在动脉粥样硬化等心血管疾病中的重要性，我们的研究结果可能为动脉粥样硬化的发病机制和治疗提供新的见解。
Objective. The present study was performed to investigate the effects and mechanisms of miR-99a on LPS-induced endothelial cell inflammation, as well as the regulation of NF-κB on miR-99a production. Methods and Results. ELISA showed that LPS treatment significantly promoted the secretion of inflammatory factors (TNF-α, IL-6, IL-1β, and MCP-1). LPS treatment also inhibited miR-99a production and promoted mTOR expression and NF-κB nuclear translocation. Overexpression of miR-99a suppressed the LPS-induced TNF-α, IL-6, IL-1β, and MCP-1 overproduction, mTOR upregulation, and NF-κB nuclear translocation. The PROMO software analysis indicated NF-κB binding site in the −1643 to −1652 region of miR-99a promoter. Dual luciferase reporter analysis, electrophoretic mobility shift assays (EMSA), and chromosome immunoprecipitation (ChIP) assays demonstrated that NF-κB promoted the transcription of miR-99a by binding to the −1643 to −1652 region of miR-99a promoter. Further studies on HUVECs verified the regulatory effects of NF-κB on miR-99a production. Conclusion. MiR-99a inhibited the LPS-induced HUVECs inflammation via inhibition of the mTOR/NF-κB signal. NF-κB promoted miR-99a production by binding to the −1643 to −1652 region of miR-99a promoter. Considering the importance of endothelial inflammation on cardiovascular diseases, such as atherosclerosis, our results may provide a new insight into the pathogenesis and therapy of atherosclerosis.