Class Ⅲ PI3K/Beclin-1自噬通路参与mmLDL上调小鼠肠系膜动脉ETA受体的研究  

The Role of ClassⅢPI3K/Beclin-1 Autophagy Pathway in Up-Regulation of ETA Receptor in Mesenteric Artery of Mice by mmLDL

作　　者：陈琛[1] 曾中三 谢希 秦旭平[1] 李洁[1,2] CHEN Chen;ZENG Zhong-san;XIE Xi;QIN Xu-ping;LI Jie(Chenzhou Frst People's Hospital,South China University,Chenzhou 423000,China;Institute of Pharmaceutical Pharmacology,Nanhua University,Hengyang 421001,China)

机构地区：[1]南华大学附属郴州市第一人民医院,湖南郴州423000 [2]南华大学药物药理研究所,湖南衡阳421001

出　　处：《中国药学杂志》2020年第11期900-907,共8页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金项目资助(81202535);湖南省教育厅创新平台开放基金项目资助(19K084);湖南省科技厅自然科学基金项目资助(2018JJ6097);湖南省卫生计生委资助课题资助(B2017183);郴州市科技局资助项目资助(zdyf201925,zdyf201821);郴州市脂质代谢紊乱诊疗技术研发中心项目资助(yfzx201907)。

摘　　要：目的探讨弱氧化性低密度脂蛋白(minimally modified low density lipoprotein,mmLDL)上调在体小鼠肠系膜动脉ETA受体的作用(endothelin type A receptors,ETA)并考察自噬是否参与这一过程。方法小鼠尾静脉注射mmLDL,腹腔注射ClassⅢPI3K自噬通路抑制剂6-氨基-3-甲基嘌呤(3-methyladenine,3-MA),探究自噬在mmLDL给药处理小鼠中的作用,微血管张力描记仪观察内皮素-1(endothelin-1,ET-1)引起的小鼠肠系膜动脉收缩量效曲线的变化,RT-PCR定量ETA受体mRNA,Western blot检测ETA受体和ClassⅢPI3K、Beclin-1、LC3-Ⅱ/Ⅰ、p62及p-NF-κB、NF-κB的蛋白水平表达。结果mmLDL引起ET-1收缩量效曲线明显增强,表现为Emax值由生理盐水(NS)组的(184.87±7.46)%上升为(319.91±20.31)%(P<0.001),pEC50值由NS组的(8.05±0.05)上升为(9.11±0.09)(P<0.01)。mmLDL在上调ClassⅢPI3K、beclin-1、LC3-Ⅱ/Ⅰ和下调p62蛋白水平的同时,也引起ETA受体mRNA水平、蛋白表达明显增加,增加了p-NF-κB的蛋白水平;腹腔注射3-MA抑制了mmLDL的这些作用。结论mmLDL能通过ClassⅢPI3K/Beclin-1通路激活自噬及下游NF-κB通路上调ETA受体。OBJECTIVE To investigate the effect of minimally modified low-density lipoprotein(mmLDL)on ETA receptor of mesenteric artery(endothelin type A receptors,ETA)in mice for the first time and whether autophagy is involved in this process.METHODS Mice were injected mmLDL in the tail vein and intraperitoneally with ClassⅢPI3K autophagy pathway inhibitor 6-amino-3-methylpurine(3-MA)to explore the role of autophagy in mmLDL treated mice.The changes of vasoconstriction curve of mesenteric artery induced by ET-1(endothelin 1)in mice were observed by a sensitive myograph system.ETA receptor was detected by RT-PCR quantitative mRNA and Western blot.The protein levels of ClassⅢPI3K,Beclin-1,LC3-Ⅱ/Ⅰ,p62 and p-NF-κB,NF-κB were detected by Western blot.RESULTS The contractility curve of ET-1 induced by mmLDL was significantly enhanced,showing that the Emax value increased from the nomal saline(NS)group(184.87±7.46)%to(319.91±20.31)%(P<0.001),the pEC50 increased from NS group(8.05±0.05)to(9.11±0.09)(P<0.01).mmLDL up-regulated ClassⅢPI3K,beclin-1,LC3-Ⅱ/Ⅰand down-regulated p62 protein level,at the same time,it also caused the ETA receptor mRNA level,protein expression increased significantly,up-regulated the protein level of p-NF-κB;intraperitoneal injection of 3-MA inhibits these effects of mmLDL.CONCLUSION mmLDL can activate autophagy and down-stream NF-κB pathway through ClassⅢPI3K/Beclin-1 pathway to up-regulate ETA receptor.

关 键 词：弱氧化性低密度脂蛋白 自噬 CLASS Ⅲ PI3K/Beclin-1信号通路 ETA受体 

分 类 号：R965[医药卫生—药理学]