基于PINK1/Parkin途径探究绿原酸对泡沫细胞线粒体自噬-炎症反应的作用机制  

Mechanism of chlorogenic acid in mitophagy and inflammation of foam cells based on PINK1/Parkin pathway

作　　者：于红红 杨韵琪 罗培[3] 俞琦 马玉玲 田维毅[1] YU Hong-hong;YANG Yun-qi;LUO Pei;YU Qi;MA Yu-ling;TIAN Wei-Yi(Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China;Center of Traditional Chinese Medicine,University of Oxford,Oxford OX12JD,Britain;Macao University of Science and Technology,Macao 999078,China)

机构地区：[1]贵州中医药大学,贵州贵阳550025 [2]牛津大学中医药研究中心,英国牛津OX12JD [3]澳门科技大学,中国澳门999078

出　　处：《中国药理学通报》2024年第5期914-920,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81860779);贵州省科技厅项目(No黔科合基础ZK[2021]549);贵州省卫生健康委科技项目(No[2021]108);贵州中医药大学研究生教育创新计划项目(No YCXKYB2023012)。

摘　　要：目的探讨绿原酸(chlorogenic acid,CGA)通过同源性磷酸酶张力蛋白诱导激酶1(PINK1)/帕金森蛋白(Parkin)信号通路对ox-LDL诱导的泡沫细胞线粒体自噬及炎症反应的调控作用。方法将RAW264.7巨噬细胞分为正常(Control)组、模型(Model)组、CGA低剂量(CGA-L)组、中剂量(CGA-M)组、高剂量(CGA-H)组、CGA-H+Mdivi-1(CGA-H+Mdi)组。油红O法鉴定细胞泡沫化;CCK-8法确定CGA干预浓度;透射电镜观察细胞线粒体结构改变;ELISA检测IL-1β、IL-18、IFN-γ表达;qRT-PCR及免疫荧光标记法检测PINK1/Parkin线粒体自噬通路相关基因及蛋白表达。结果油红O染色显示泡沫细胞模型制备成功。与Model组相比,不同浓度CGA干预后细胞线粒体损伤明显减轻,诱导了PINK1、Parkin、p-Parkin、PHB2、LC3Ⅱ表达,抑制了TOMM20表达,以及降低了IL-1β、IL-18、IFN-γ的表达(P<0.05或P<0.01);与CGA-H组相比,CGA-H+Mdi组细胞线粒体损伤明显加重,抑制了PINK1、Parkin、p-Parkin、PHB2、LC3Ⅱ表达,诱导了TOMM20表达,以及增强了IL-1β、IL-18、IFN-γ表达(P<0.01)。结论CGA可通过调控PINK1/Parkin通路诱导泡沫细胞线粒体自噬以及抑制了促炎因子IL-1β、IL-18、IFN-γ过表达,这可能是CGA抗动脉粥样硬化机制之一。Aim To investigate the regulatory effects of chlorogenic acid(CGA)on mitochondrial autophagy and inflammation in ox-LDL induced foam cells through PINK1/Parkin signaling pathway.Methods RAW264.7 macrophages were divided into the control group,model group,CGA-L group,CGA-M group,CGA-H group and CGA-H+Mdi group.Oil red O method was used to identify cell foam.The intervention concentration of CGA was determined by CCK-8 method.The mitochondrial structure was observed by transmission electron microscope.The expression of IL-1β,IL-18 and IFN-γwas detected by ELISA.The expression of genes and proteins associated with PINK1/Parkin mitochondrial autophagy pathway was detected by qRT-PCR and immunofluorescence labeling.Results Oil red O staining showed that the foam cell model was successfully prepared.Compared with the model group,mitochondrial damage was significantly reduced after CGA intervention at different concentrations,and the expressions of PINK1,Parkin,p-Parkin,PHB2 and LC3Ⅱwere induced,while the expression of TOMM20 was inhibited.The expressions of IL-1β,IL-18 and IFN-γdecreased(P<0.05 or P<0.01).Compared with the CGA-H group,the CGA-H+Mdi group significantly increased mitochondrial damage,inhibited the expression of PINK1,Parkin,p-Parkin,PHB2,LC3Ⅱ,induced the expression of TOMM20,and enhanced the expression of IL-1β,IL-18,IFN-γ(P<0.01).Conclusions CGA can induce mitochondrial autophagy in foam cells by regulating PINK1/Parkin pathway and inhibit the overexpression of pro-inflammatory factors IL-1β,IL-18 and IFN-γ,which may be one of the anti-atherosclerosis mechanisms of CGA.

关 键 词：绿原酸 泡沫细胞 PINK1/Parkin 线粒体自噬 炎症反应 动脉粥样硬化 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R364.5[医药卫生—基础医学] R392R543.5R916.4