长链非编码RNA LOC286367-ABCA1途径介导异丙酚抗炎作用的新机制研究

It has been shown that propofol has anti-inflammatory effect as a kind of widely used agent for clinical anesthesia. ATP-binding cassette transporter A1 (ABCA1) is a member of a family of highly conserved transmembrane transport proteins and its expression can be up-regulated by propofol. Our group has recently found that ABCA1 has anti-inflammatory properties. However, it is unclear that whether propofol could exert its anti-inflammatory activity by regulation of ABCA1 and the possible mechanisms involving in this progress has not been fully explored. Microarray analysis reveals that expression of long non-coding RNA (lncRNA) LOC286367 is down-regulated by treatment with propofol in macrophages. In addition, bioinformatic analysis shows that the lncRNA LOC286367 and ABCA1 are located in the same chromosome. Furthermore, ABCA1 expression is inhibited by overexpression of LOC286367 in macrophages. Thus, we suppose that propofol may first affect expression and function of ABCA1 by regulating expression of LOC286367 and then exert its anti-inflammatory actions. In this project, we aim to explore the molecular mechanisms responsible for decreasing LOC286367 expression in response to propofol and to investigate the regulatory mechanism responsible for reducing ABCA1 expression in response to LOC286367, and to observe the effects of propofol on inflammation by regulation of LOC286367-ABCA1 pathway. Therefore, our study may be useful in understanding the critical effects of propofol on inflammation and provide new scientific evidence to establish LOC286367-ABCA1 pathway as a new target for regulation of inflammation.

异丙酚作为临床常用麻醉药具有抗炎作用，本课题组最新研究发现ABCA1作为细胞跨膜蛋白具有抗炎作用且异丙酚能上调ABCA1表达。但异丙酚是否可以通过ABCA1发挥抗炎作用及相关机制并不清楚。我们前期预实验通过基因芯片发现异丙酚能抑制lncRNA LOC286367的表达，生物信息学分析和预实验结果表明LOC286367与ABCA1定位于同一染色体且LOC286367可抑制ABCA1表达。因此我们首次提出异丙酚可能通过抑制LOC286367表达从而上调ABCA1表达进而发挥抗炎作用。本项目拟探讨异丙酚抑制LOC286367的具体机制，探讨LOC286367调控ABCA1的具体机制，通过增强LOC286367和/或ABCA1表达，观察LOC286367-ABCA1介导异丙酚的抗炎效果，为深入阐述异丙酚的抗炎作用拓展新思路，为LOC286367-ABCA1途径作为调控炎症反应新靶点提供科学依据。

异丙酚作为临床常用麻醉药具有抗炎作用，本课题组最新研究发现ABCA1作为细胞跨膜蛋白具有抗炎作用且异丙酚能上调ABCA1表达。但异丙酚是否可以通过ABCA1发挥抗炎作用及相关机制并不清楚。我们前期预实验通过基因芯片发现异丙酚能抑制lncRNA LOC286367的表达，生物信息学分析和预实验结果表明LOC286367与ABCA1定位于同一染色体且LOC286367可抑制ABCA1表达。采用实时定量PCR和免疫印迹对基因芯片结果进行验证，我们发现THP-1巨噬细胞经异丙酚处理后其LOC286367表达明显下调而ABCA1的表达明显上调；采用慢病毒转染方式在THP-1巨噬细胞中过表达LOC286367后能明显抑制ABCA1表达；采用ELISA法分析LOC286367对THP-1巨噬细胞炎症因子表达的影响，我们发现LOC286367明显促进巨噬细胞炎症因子的分泌。构建lncRNA LOC286367过表达载体转染THP-1巨噬细胞，经LPS诱导的THP-1巨噬细胞中TNF-α、IL-1β及IL-6 的含量均明显上升；将LV-LOC286367注入进入小鼠的尾静脉，经ELISA检测血清中炎性细胞因子的含量，结果发现LV- LOC286367转染后小鼠血清中TNF-α、IL-1β、IL-6的含量均明显上升，说明lncRNA LOC286367参与了异丙酚对THP-1巨噬细胞和小鼠经LPS诱导下炎性细胞因子表达的调控。本研究结果提示丙泊酚通过促进ABCA1表达降低LPS诱导的THP-1巨噬细胞促炎细胞因子（包括TNF-α、IL-1β和IL-6）的表达水平来抑制炎症反应，这是以LOC286367依赖的方式实现的。这些发现为深入阐述异丙酚的抗炎作用拓展新思路，为LOC286367-ABCA1途径作为调控炎症反应新靶点提供科学依据。