title: The Protective Role of Autophagy in Matrix Metalloproteinase-Mediated Cell Transmigration and Cell Death in High-Glucose-Treated Endothelial Cells.
abstract: Diabetes mellitus may cause vascular endothelial damage via endothelial matrix metalloproteinase-2 (MMP-2). The role of endothelial autophagy in MMP-2-mediated cell injury in response to high-glucose (HG) stimulation was rarely described. In this study, we used HG-treated human umbilical vein endothelial cells (HUVECs) to investigate the effect of autophagy on MMP-2-induced cell transmigration and apoptosis. THP-1 transmigration was detected by the transmigration assay. Light chain 3 (LC3, representing autophagy), MMP-2, and poly (ADP-ribose) polymerase (PARP, representing apoptosis) of HG (33 mM)-treated HUVECs were evaluated by western blot analysis. The MMP-2 activity was also examined by gelatin zymography. We used GM6001 (10 μM, an MMP-2 inhibitor) to investigate the relationship of MMP-2 and THP-1 transmigration. Using 3-methyladenine (3MA, 5 mM, an LC3 inhibitor), we explored the effects of autophagy on MMP-2 expression, THP-1 transmigration, and apoptosis. Our results showed that HG increased LC3-II expression, MMP-2 activity, THP-1 transmigration, and cleaved PARP expression in a time-dependent manner (0-48 h); among them, LC3-II appeared earlier (0-24 h) than the others (24-48 h). GM6001 suppressed MMP-2 activity and ameliorated THP-1 transmigration. 3MA suppressed LC3-II expression and increased MMP-2 expression, THP-1 transmigration, and cleaved PARP expression. From these sequential findings, we demonstrated that autophagy plays a protective role in MMP-2-mediated cell transmigration and cell death in HG-stimulated HUVECs.
authors: Division of Cardiology, Department of Internal Medicine, National Taiwan University College of Medicine and National Taiwan University Hospital, Taipei, 10002, Taiwan.
authors: Chia-Lun
authors: CL
authors: Chao
authors: Division of Cardiology, Department of Internal Medicine, Ministry of Health and Welfare, Executive Yuan, Taoyuan General Hospital, 1492 Chung-Shan Road, Taoyuan, 33004, Taiwan.
authors: Chun-Pin
authors: CP
authors: Chuang
authors: Division of Cardiology, Department of Internal Medicine, Ministry of Health and Welfare, Executive Yuan, Taoyuan General Hospital, 1492 Chung-Shan Road, Taoyuan, 33004, Taiwan.
authors: Yen-Fen
authors: YF
authors: Cheng
authors: R&D Center for Membrane Technology, Department of Chemical Engineering, Chung Yuan Christian University, Taoyuan, 32023, Taiwan.
authors: Kueir-Rarn
authors: KR
authors: Lee
authors: R&D Center for Membrane Technology, Department of Chemical Engineering, Chung Yuan Christian University, Taoyuan, 32023, Taiwan.
authors: Yung
authors: Y
authors: Chang
authors: Department of Physical Medicine and Rehabilitation, Ministry of Health and Welfare, Executive Yuan, Taoyuan General Hospital, Taoyuan, 33004, Taiwan.
authors: Shun-Ping
authors: SP
authors: Cheng
authors: Division of Cardiology, Department of Internal Medicine, Far Eastern Memorial Hospital, New Taipei City, 22060, Taiwan.
authors: Wan-Khey
authors: WK
authors: Chan
authors: Division of Cardiology, Department of Internal Medicine, Ministry of Health and Welfare, Executive Yuan, Taoyuan General Hospital, 1492 Chung-Shan Road, Taoyuan, 33004, Taiwan. HoFM888@yahoo.com.tw.|R&D Center for Membrane Technology, Department of Chemical Engineering, Chung Yuan Christian University, Taoyuan, 32023, Taiwan. HoFM888@yahoo.com.tw.|Cardiovascular Research Center, College of Medicine, Taipei Medical University, Taipei, 11031, Taiwan. HoFM888@yahoo.com.tw.
authors: Feng-Ming
authors: FM
authors: Ho
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives / Q000378:metabolism; D017209:Apoptosis / Q000502:physiology*; D001343:Autophagy / Q000502:physiology*; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology*; D003920:Diabetes Mellitus / Q000473:pathology; D004151:Dipeptides / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology / Q000473:pathology; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology*; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism*; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D061965:Matrix Metalloproteinase Inhibitors / Q000494:pharmacology; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism
chemical_list: D004151:Dipeptides; D061965:Matrix Metalloproteinase Inhibitors; C078131:N-(2(R)-2-(hydroxamidocarbonylmethyl)-4-methylpentanoyl)-L-tryptophan methylamide; C025946:3-methyladenine; D011065:Poly(ADP-ribose) Polymerases; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2; D005947:Glucose; D000225:Adenine
keywords: autophagy; endothelial cells; high glucose; matrix metalloproteinase; transmigration