GAPDH-silence preserves H9C2 cells from acute hypoxia and reoxygenation injury.
BACKGROUND
Acute hypoxia and reoxygenation injury, as a common environmental stress condition, is a basic condition of most pathophysiological processes. It has been approve that autophagy and oxidant stress could contribute to acute hypoxia and reoxygenation injury. This study is aimed to examine the effect of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) silence on cell injury with acute hypoxia and reoxygenation injury by autophagy and antioxidant stress pathway.


METHODS
GAPDH expression was silenced by siRNA in H9C2 cardiomyoblasts with acute hypoxia and reoxygenation injury. Autophagy was detected by western blot for autophagy proteins and monodansylcadaverine (MDC) staining for acidic substances. Pro-apoptosis protein and flow cytometry were used to assess cell apoptosis and death and intracellular adenosine triphosphate (ATP) relative concentration was measured. Oxidant stress was assessed by measuring 2'-7'-dichlorodihydrofluorescein diacetate (DCFH-DA), thiobarbituric acid reactive substances (TBARS), glutathione (GSH) and super oxide dismutase (SOD).


RESULTS
In this study, GAPDH-silence enhanced autophagy in H9C2 cells with acute hypoxia and reoxygenation injury, decreased oxidant stress and increased antioxidant pathways; and reduced cell apoptosis and death. However, GAPDH-silence had no significant effect on cell energy.


CONCLUSIONS
GAPDH pre-silence by siRNA reduces H9C2 cell death occurring via autophagy and anti-oxidative stress pathway in acute hypoxia and reoxygenation injury. This study enriches the understanding of GAPDH pathophysiology role, and provides potential new therapeutic targets for cardiac disease states characterized by oxidative stress.
Department of Cardiology, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, China. Electronic address: shaoliang021224@hotmail.com.
Shao
S
Liang
Department of Cardiovascular Medicine, First Affiliated Hospital of Medical College of Xi'an Jiaotong University, Xi'an, Shaanxi Province, China.
Ma
M
Aiqun
North Shore Heart Research Group, Kolling Institute of Medical Research, University of Sydney, Sydney, Australia; Department of Cardiology, Royal North Shore Hospital, Sydney, Australia.
Gemma
G
Figtree
Department of Geriatrics & Neurology, The Second Affiliated Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, China. Electronic address: zhangkiki520@126.com.
Zhang
Z
Ping
D000975:Antioxidants / Q000378:metabolism; D001343:Autophagy / Q000235:genetics; D015687:Cell Hypoxia / Q000235:genetics; D002460:Cell Line; D004734:Energy Metabolism / Q000235:genetics; D015870:Gene Expression; D020868:Gene Silencing; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D000860:Hypoxia / Q000235:genetics* / Q000378:metabolism; D032386:Myoblasts, Cardiac / Q000378:metabolism; D018384:Oxidative Stress / Q000235:genetics; D017382:Reactive Oxygen Species / Q000378:metabolism; D015427:Reperfusion Injury / Q000235:genetics* / Q000378:metabolism
D000975:Antioxidants; D017382:Reactive Oxygen Species; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases
Autophagy; GAPDH; Oxidative stress