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目的 探究葛根素是否通过线粒体自噬通路调控巨噬细胞极化,进而改善动脉粥样硬化。
方法 以ApoE-/-小鼠高脂饮食构建动脉粥样硬化模型,葛根素干预12周后采用苏木精-伊红(hematoxylin-eosin, HE)染色法和油红O (oil red O, ORO)染色法观察主动脉斑块;蛋白质免疫印迹检测(western blot, WB)蛋白表达。以RAW264.7巨噬细胞构建M1极化模型,实时荧光定量聚合酶链反应检测和WB验证巨噬细胞极化表型及线粒体自噬相关蛋白表达,透射电镜观察线粒体自噬形态。
结果 葛根素显著减小主动脉斑块面积,降低脂质沉积;上调主动脉组织磷酸酶张力蛋白同源物诱导的激酶1 (PTEN-induced kinase 1, Pink1)、帕金蛋白(Parkin E3泛素连接酶)(Parkin RBR E3 ubiquitin protein ligase, Parkin)蛋白表达及微管相关蛋白1轻链3-Ⅱ/Ⅰ(microtubule-associated protein 1 light chain 3-Ⅱ/Ⅰ, LC3 Ⅱ/Ⅰ),下调隔离体蛋白1 (sequestosome 1, p62);抑制M1型,促进M2型基因表达。而沉默Pink1后,葛根素对巨噬细胞极化的调控作用及抗炎效应显著减弱,线粒体自噬激活受阻。
结论 葛根素可激活由Pink1/Parkin所介导的线粒体自噬通路,使巨噬细胞向抑制炎症方向极化(M2),改善动脉粥样硬化。本研究通过线粒体自噬调控巨噬细胞极化,丰富了葛根素抗AS的作用机制,并为靶向Pink1/Parkin通路干预其炎症进展提供实验依据和潜在治疗思路。
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