Thrombin-induced autophagy: a potential role in intracerebral hemorrhage.
Autophagy occurs in the brain after intracerebral hemorrhage (ICH) and thrombin contributes to ICH-induced brain injury and cell death. In this study, we investigated whether thrombin may activate autophagy (in vivo and in cultured astrocytes) and its potential role in ICH. Autophagy was examined using electron microscopy, conversion of light chain 3(LC3) from the LC3-I form to LC3-II, cathepsin D Western blotting and monodansylcadaverine (MDC) staining to detect autophagic vacuoles. 3-Methyladenine (3-MA) was used as an autophagy inhibitor. In vivo, we found that intracaudate injection of thrombin increased conversion of LC3-I to LC3-II, cathepsin D levels, and formation of autophagic vacuoles in the ipsilateral basal ganglia. ICH-induced upregulation of LC3-I to LC3-II conversion and cathepsin D levels was reduced by a thrombin inhibitor, hirudin. In cultured astrocytes, thrombin enhanced the conversion of LC3-I to LC3-II and increased MDC-labeled autophagic vacuoles. 3-MA inhibited thrombin-induced autophagic vacuole formation and exacerbated thrombin-induced cell death. These results indicate that thrombin activates autophagy in the brain and that thrombin has a role in ICH-induced autophagy.
Department of Neurosurgery, University of Michigan, Ann Arbor, Michigan, USA.
Shukun
S
Hu
Guohua
G
Xi
Hang
H
Jin
Yangdong
Y
He
Richard F
RF
Keep
Ya
Y
Hua
D000818:Animals; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism*; D001343:Autophagy / Q000187:drug effects / Q000502:physiology*; D015153:Blotting, Western; D002543:Cerebral Hemorrhage / Q000378:metabolism*; D008297:Male; D046529:Microscopy, Electron, Transmission; D051381:Rats; D017207:Rats, Sprague-Dawley; D013917:Thrombin / Q000378:metabolism* / Q000494:pharmacology
D013917:Thrombin