Atherosclerosis is a progressive, chronic inflammation in arterial walls. Long noncoding RNAs (lncRNAs) participate in inflammation, but the exact mechanism in atherosclerosis is unclear. Our microarray analyses revealed that the levels of lncRNA-FA2H-2 were significantly decreased by oxidized low-density lipoprotein (OX-LDL). Bioinformatics analyses indicated that mixed lineage kinase domain-like protein (MLKL) might be regulated by lncRNA-FA2H-2. In vitro experiments showed that lncRNA-FA2H-2 interacted with the promoter of the MLKL gene, downregulated MLKL expression, and the binding sites between -750 and 471 were necessary for lncRNA-FA2H-2 responsiveness to MLKL. Silencing lncRNA-FA2H-2 and overexpression of MLKL could activate inflammation and inhibited autophagy flux. Both lncRNA-FA2H-2 knockdown and overexpression of MLKL could significantly aggravate inflammatory responses induced by OX-LDL. We found that the 3-methyladenine (3-MA) and Atg7-shRNA enhanced inflammatory responses induced by knockdown of lncRNA-FA2H-2 and overexpression of MLKL. We demonstrated that the effects of MLKL on autophagy might be associated with a mechanistic target of rapamycin (mTOR)-dependent signaling pathways. In vivo experiments with apoE knockout mice fed a western diet demonstrated that LncRNA-FA2H-2 knockdown decreased microtubule-associated expression of microtubule-associated protein 1 light chain 3 II and lysosome-associated membrane protein 1, but increased expression of sequestosome 1 (p62), MLKL, vascular cell adhesion molecule-1, monocyte chemoattractant protein-1, and interleukin-6 in atherosclerotic lesions. Our findings indicated that the lncRNA-FA2H-2-MLKL pathway is essential for regulation of autophagy and inflammation, and suggested that lncRNA-FA2H-2 and MLKL could act as potential therapeutic targets to ameliorate atherosclerosis-related diseases.

中文翻译：

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LncRNA-FA2H-2-MLKL途径在动脉粥样硬化中的作用是通过依赖于mTOR的信号传导调节自噬通量和炎症。

动脉粥样硬化是动脉壁的进行性慢性炎症。长的非编码RNA（lncRNA）参与炎症，但动脉粥样硬化的确切机制尚不清楚。我们的微阵列分析表明，氧化型低密度脂蛋白（OX-LDL）显着降低了lncRNA-FA2H-2的水平。生物信息学分析表明，混合谱系激酶域样蛋白（MLKL）可能受lncRNA-FA2H-2调控。体外实验表明，lncRNA-FA2H-2与MLKL基因的启动子相互作用，下调了MLKL表达，并且-750和471之间的结合位点对于lncRNA-FA2H-2对MLKL的响应是必需的。沉默lncRNA-FA2H-2和过度表达MLKL可激活炎症并抑制自噬通量。lncRNA-FA2H-2的敲低和MLKL的过表达均可显着加重OX-LDL诱导的炎症反应。我们发现3-甲基腺嘌呤（3-MA）和Atg7-shRNA增强了由lncRNA-FA2H-2的敲低和MLKL的过表达诱导的炎症反应。我们证明了MLKL对自噬的影响可能与雷帕霉素（mTOR）依赖的信号通路的机械目标有关。载有西方饮食的apoE敲除小鼠的体内实验表明，LncRNA-FA2H-2敲低可降低微管相关蛋白1轻链3 II和溶酶体相关膜蛋白1的微管相关表达，但可增加螯合物1（p62 ），MLKL，血管细胞粘附分子1，单核细胞趋化蛋白1和白细胞介素6在动脉粥样硬化病变中的作用。