祛痰清瘀方通过活化CREB上调自噬缓解oxLDL对人主动脉内皮细胞的致炎作用.docxVIP

? ? 祛痰清瘀方通过活化CREB上调自噬缓解oxLDL对人主动脉内皮细胞的致炎作用 ? ? Summary　目的：探讨在二陈汤和桃红四物汤基础上化裁而成的祛痰清瘀方通过活化CREB上调自噬抑制氧化低密度脂蛋白（oxLDL）对人主动脉内皮细胞（HAEC）的致炎作用及机制。方法：构建oxLDL诱导HAEC炎症模型，经相应含药血清处理，应用ELISA定量培养上清液中IL-1β、IL-6和IL-8的含量，Western blot法检测HAEC中IκBα、NF-κB p65和CREB的表达及p65和CREB的核转位，HAEC自噬活性及自噬组件蛋白PIK3C3、Beclin1、Atg7和Atg14的表达，以定量PCR评价HAEC自噬相关基因PIK3C3、BECN1、ATG7和ATG14的转录活性，并通过操控CREB表达影响自噬，以研究祛痰清瘀方的作用机制。结果：与oxLDL组相比，祛痰清瘀方能够显著抑制oxLDL诱导HAEC分泌IL-1β、IL-6和IL-8的作用，减少p65蛋白水平，阻止其核转位（P0.01）；祛痰清瘀方能够诱导CREB磷酸化及核转位，上调HAEC自噬相关基因PIK3C3、BECN1、ATG7和ATG14的转录及蛋白表达，诱导HAEC自噬（P0.01），从而介导p65经自噬途径降解。结论：祛痰清瘀方通过激活CREB上调HAEC自噬，介导p65降解，减轻oxLDL诱导的HAEC炎症反应。 Keys　祛痰清瘀方；动脉粥样硬化；人主动脉内皮细胞；炎症 Qutan Qingyu decoction ameliorates oxLDL-induced inflammation in human aortic endothelial cells by activating CREB and up-regulating autophagy KOU Xin-hui, CHEN Jun, CUI Xiang, ZHAO Heng-xia, LI Hui-lin* (Shenzhen Traditional Chinese Medicine Hospital, Shenzhen 518033) Abstract　Objective: To study the effect and mechanism of Qutan Qingyu decoction on oxidized low-density lipoprotein (oxLDL)-induced human aortic endothelial cell (HAEC) inflammation by activating CREB and up-regulating autophagy. Methods: The oxLDL-induced inflammatory model of HAEC was established. The concentrations of IL-1β, IL-6 and IL-8 in the culture supernatant of HAEC were quantified by ELISA. The protein levels of IκBα, p65, CREB, and the autophagic activity were determined by Western blot. The expression levels of autophagy-related genes were detected by qPCR. Results: Compared with oxLDL group, Qutan Qingyu decoction could significantly inhibit the production of IL-1β, IL-6 and IL-8, compromise the expression of p65 and its nuclear translocation in HAEC (P 0.01). Qutan Qingyu decoction could induce CREB phosphorylation, promote nuclear translocation, and up-regulate the expression of autophagy related genes PIK3C3, BECN1, ATG7, and ATG14 in HAEC (P 0.01). Qutan Qingyu decoction could induce autophagy, and thus mediate the degradation of p65 through autophagy i