Aldehyde dehydrogenase 2 protects human umbilical vein endothelial cells against oxidative damage and increases endothelial nitric oxide production to reverse nitroglycerin tolerance.
Medical nitroglycerin (glyceryl trinitrate, GTN) use is limited principally by tolerance typified by a decrease in nitric oxide (NO) produced by biotransformation. Such tolerance may lead to endothelial dysfunction by inducing oxidative stress. In vivo studies have demonstrated that aldehyde dehydrogenase 2 (ALDH2) plays important roles in GTN biotransformation and tolerance. Thus, modification of ALDH2 expression represents a potentially effective strategy to prevent and reverse GTN tolerance and endothelial dysfunction. In this study, a eukaryotic expression vector containing the ALDH2 gene was introduced into human umbilical vein endothelial cells (HUVECs) by liposome-mediated transfection. An indirect immunofluorescence assay showed that ALDH2 expression increased 24 h after transfection. Moreover, real-time polymerase chain reaction and western blotting revealed significantly higher ALDH2 mRNA and protein expression in the gene-transfected group than in the two control groups. GTN tolerance was induced by treating HUVECs with 10 mM GTN for 16 h + 10 min, which significantly decreased NO levels in control cells, but not in those transfected with ALDH2. Overexpression of ALDH2 increased cell survival against GTN-induced cytotoxicity and conferred protection from oxidative damage resulting from nitrate tolerance, accompanied by decreased production of intracellular reactive oxygen species and reduced expression of heme oxygenase 1. Furthermore, ALDH2 overexpression promoted Akt phosphorylation under GTN tolerance conditions. ALDH2 gene transfection can reverse and prevent tolerance to GTN through its bioactivation and protect against oxidative damage, preventing the development of endothelial dysfunction.
Department of Hematology, Affiliated Hospital of GuiZhou Medical University, Guiyang, China.|Key Laboratory of Hematological Disease Diagnostic & Treat Centre of GuiZhou Province, Guiyang, China.|GuiZhou Province Hematopoietic Stem Cell Transplantation Center, Affiliated Hospital of GuiZhou Medical University, Guiyang, China.
X Y
XY
Hu
Department of Pharmacy, Affiliated BaiYun Hospital of GuiZhou Medical University, China.|Department of Pharmacy, Affiliated Hospital of GuiZhou Medical University, Guiyang, China.
Q
Q
Fang
Key Laboratory of Hematological Disease Diagnostic & Treat Centre of GuiZhou Province, Guiyang, China.|Department of Pharmacy, Affiliated BaiYun Hospital of GuiZhou Medical University, China.|Department of Pharmacy, Affiliated Hospital of GuiZhou Medical University, Guiyang, China.
D
D
Ma
Department of Pharmacy, Affiliated BaiYun Hospital of GuiZhou Medical University, China.
L
L
Jiang
Key Laboratory of Hematological Disease Diagnostic & Treat Centre of GuiZhou Province, Guiyang, China.|GuiZhou Province Hematopoietic Stem Cell Transplantation Center, Affiliated Hospital of GuiZhou Medical University, Guiyang, China.
Y
Y
Yang
School of Pharmacy, GuiZhou Medical University, Guiyang, China.
J
J
Sun
School of Pharmacy, GuiZhou Medical University, Guiyang, China.
C
C
Yang
Department of Hematology, Affiliated Hospital of GuiZhou Medical University, Guiyang, China.|Key Laboratory of Hematological Disease Diagnostic & Treat Centre of GuiZhou Province, Guiyang, China.|GuiZhou Province Hematopoietic Stem Cell Transplantation Center, Affiliated Hospital of GuiZhou Medical University, Guiyang, China.
J S
JS
Wang
D000071396:Aldehyde Dehydrogenase, Mitochondrial / Q000096:biosynthesis* / Q000235:genetics; D004361:Drug Tolerance / Q000235:genetics*; D042783:Endothelial Cells / Q000187:drug effects / Q000473:pathology; D005786:Gene Expression Regulation / Q000187:drug effects; D051547:Heme Oxygenase-1 / Q000096:biosynthesis* / Q000235:genetics; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D006801:Humans; D009569:Nitric Oxide / Q000235:genetics / Q000378:metabolism; D005996:Nitroglycerin / Q000009:adverse effects* / Q000627:therapeutic use; D018384:Oxidative Stress / Q000187:drug effects; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis
D012333:RNA, Messenger; D009569:Nitric Oxide; D051547:Heme Oxygenase-1; C096127:ALDH2 protein, human; D000071396:Aldehyde Dehydrogenase, Mitochondrial; D051057:Proto-Oncogene Proteins c-akt; D005996:Nitroglycerin