Trentzsch Marcus, Nyamugenda Eugene, Miles Tiffany K, Griffin Haven, Russell Susan, Koss Brian, Cooney Kimberly A, Phelan Kevin D, Tackett Alan J, Iyer Srividhya, Boysen Gunnar, Baldini Giulia
1Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, Little Rock, AR USA.
2Department of Neurobiology & Developmental Sciences, University of Arkansas for Medical Sciences, Little Rock, AR USA.
Cell Death Discov. 2020 Feb 18;6:8. doi: 10.1038/s41420-020-0241-z. eCollection 2020.
Genetic obesity increases in liver phosphatidylcholine (PC)/phosphatidylethanolamine (PE) ratio, inducing endoplasmic reticulum (ER) stress without concomitant increase of ER chaperones. Here, it is found that exposing mice to a palm oil-based high fat (HF) diet induced obesity, loss of liver PE, and loss of the ER chaperone Grp78/BiP in pericentral hepatocytes. In Hepa1-6 cells treated with elevated concentration of palmitate to model lipid stress, Grp78/BiP mRNA was increased, indicating onset of stress-induced Unfolded Protein Response (UPR), but Grp78/BiP protein abundance was nevertheless decreased. Exposure to elevated palmitate also induced in hepatoma cells decreased membrane glycosylation, nuclear translocation of pro-apoptotic C/EBP-homologous-protein-10 (CHOP), expansion of ER-derived quality control compartment (ERQC), loss of mitochondrial membrane potential (MMP), and decreased oxidative phosphorylation. When PE was delivered to Hepa1-6 cells exposed to elevated palmitate, effects by elevated palmitate to decrease Grp78/BiP protein abundance and suppress membrane glycosylation were blunted. Delivery of PE to Hepa1-6 cells treated with elevated palmitate also blunted expansion of ERQC, decreased nuclear translocation of CHOP and lowered abundance of reactive oxygen species (ROS). Instead, delivery of the chemical chaperone 4-phenyl-butyrate (PBA) to Hepa1-6 cells treated with elevated palmitate, while increasing abundance of Grp78/BiP protein and restoring membrane glycosylation, also increased ERQC, expression and nuclear translocation of CHOP, non-mitochondrial oxygen consumption, and generation of ROS. Data indicate that delivery of PE to hepatoma cells under lipid stress recovers cell function by targeting the secretory pathway and by blunting pro-apoptotic branches of the UPR.
遗传性肥胖会使肝脏中磷脂酰胆碱(PC)/磷脂酰乙醇胺(PE)的比例升高,引发内质网(ER)应激,而内质网伴侣蛋白却不会随之增加。在此研究中,发现给小鼠喂食基于棕榈油的高脂(HF)饮食会导致肥胖、肝脏PE流失以及中央周围肝细胞中内质网伴侣蛋白Grp78/BiP的流失。在用高浓度棕榈酸处理Hepa1-6细胞以模拟脂质应激的实验中,Grp78/BiP mRNA增加,表明应激诱导的未折叠蛋白反应(UPR)开始,但Grp78/BiP蛋白丰度却下降。暴露于高浓度棕榈酸还会导致肝癌细胞中膜糖基化减少、促凋亡的C/EBP同源蛋白10(CHOP)核转位、内质网衍生的质量控制区室(ERQC)扩张、线粒体膜电位(MMP)丧失以及氧化磷酸化减少。当将PE递送至暴露于高浓度棕榈酸的Hepa1-6细胞时,高浓度棕榈酸降低Grp78/BiP蛋白丰度和抑制膜糖基化的作用会减弱。将PE递送至用高浓度棕榈酸处理的Hepa1-6细胞也会减弱ERQC的扩张、降低CHOP的核转位并降低活性氧(ROS)的丰度。相反,将化学伴侣4-苯基丁酸盐(PBA)递送至用高浓度棕榈酸处理的Hepa1-6细胞,在增加Grp78/BiP蛋白丰度并恢复膜糖基化的同时,还会增加ERQC、CHOP的表达和核转位、非线粒体氧消耗以及ROS的生成。数据表明,在脂质应激下将PE递送至肝癌细胞可通过靶向分泌途径并减弱UPR的促凋亡分支来恢复细胞功能。