title: Advanced Glycation End Products Inhibit the Proliferation of Human Umbilical Vein Endothelial Cells by Inhibiting Cathepsin D.
abstract: We aimed to investigate the effect of advanced glycation end products (AGEs) on the proliferation and migration ability of human umbilical vein endothelial cells (HUVECs). Cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay, real-time cell analyzer and 5-Ethynyl-2'-deoxyuridine (EdU) staining. Cell migration was detected by wound-healing and transwell assay. AGEs significantly inhibited the proliferation and migration of HUVECs in a time-and dose-dependent way. Western blotting revealed that AGEs dramatically increased the expression of microtubule-associated protein 1 light chain 3 (LC3) II/I and p62. Immunofluorescence of p62 and acridine orange staining revealed that AGEs significantly increased the expression of p62 and the accumulation of autophagic vacuoles, respectively. Chloroquine (CQ) could further promote the expression of LC3 II/I and p62, increase the accumulation of autophagic vacuoles and promote cell injury induced by AGEs. In addition, AGEs reduced cathepsin D (CTSD) expression in a time-dependent way. Overexpression of wild-type CTSD significantly decreased the ratio of LC 3 II/I as well as p62 accumulation induced by AGEs, but overexpression of catalytically inactive mutant CTSD had no such effects. Only overexpression of wild-type CTSD could restore the proliferation of HUVECs inhibited by AGEs. However, overexpression of both wild-type CTSD and catalytically inactive mutant CTSD could promote the migration of HUVECs inhibited by AGEs. Collectively, our study found that AGEs inhibited the proliferation and migration in HUVECs and promoted autophagic flux, which in turn played a protective role against AGEs-induced cell injury. CTSD, in need of its catalytic activity, may promote proliferation in AGEs-treated HUVECs independent of the autophagy-lysosome pathway. Meanwhile, CTSD could improve the migration of AGEs-treated HUVECs regardless of its enzymatic activity.
authors.affiliation: Department of Cardiology, The People's Hospital of Liaoning Province, Shenyang 110001, China. xi.aohan1989@163.com.|Department of Cardiology, The People's Hospital of China Medical University, Shenyang 110001, China. xi.aohan1989@163.com.|Department of Cardiology, The First Hospital of China Medical University, Shenyang 110001, China. xi.aohan1989@163.com.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Cardiology, The First Hospital of China Medical University, Shenyang 110001, China. chang.ye@stu.xjtu.edu.cn.
authors.forename: Ye
authors.initials: Y
authors.lastname: Chang
authors.affiliation: Department of Cardiology, The First Hospital of China Medical University, Shenyang 110001, China. yening_cmu@163.com.
authors.forename: Ning
authors.initials: N
authors.lastname: Ye
authors.affiliation: Department of Cardiology, The First Hospital of China Medical University, Shenyang 110001, China. 18240409506@163.com.
authors.forename: Dongxue
authors.initials: D
authors.lastname: Dai
authors.affiliation: Department of Cardiology, The First Hospital of China Medical University, Shenyang 110001, China. pinkrainsky@foxmail.com.
authors.forename: Yintao
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Cardiology, The First Hospital of China Medical University, Shenyang 110001, China. zhangnj1989@126.com.
authors.forename: Naijin
authors.initials: N
authors.lastname: Zhang
authors.affiliation: Department of Cardiology, The First Hospital of China Medical University, Shenyang 110001, China. gzhsun66@163.com.
authors.forename: Guozhe
authors.initials: G
authors.lastname: Sun
authors.affiliation: Department of Cardiology, The First Hospital of China Medical University, Shenyang 110001, China. yxsun@mail.cmu.edu.cn.
authors.forename: Yingxian
authors.initials: Y
authors.lastname: Sun
mesh_terms: D001343:Autophagy / Q000187:drug effects; D002402:Cathepsin D / Q000037:antagonists & inhibitors*; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D017127:Glycation End Products, Advanced / Q000494:pharmacology*; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology*; D006801:Humans
chemical_list: D017127:Glycation End Products, Advanced; D002402:Cathepsin D
keywords: advanced glycation end products (AGEs); autophagy; cathepsin D (CTSD); human umbilical vein endothelial cells (HUVECs); migration; proliferation