Necrostatin-1 suppresses autophagy and apoptosis in mice traumatic brain injury model.
Traumatic brain injury (TBI) results in neuronal apoptosis, autophagic cell death and necroptosis. Necroptosis is a newly discovered caspases-independent programmed necrosis pathway which can be triggered by activation of death receptor. Previous works identified that necrostatin-1 (NEC-1), a specific necroptosis inhibitor, could reduce tissue damage and functional impairment through inhibiting of necroptosis process following TBI. However, the role of NEC-1 on apoptosis and autophagy after TBI is still not very clear. In this study, the amount of TBI-induced neural cell deaths were counted by PI labeling method as previously described. The expression of autophagic pathway associated proteins (Beclin-1, LC3-II, and P62) and apoptotic pathway associated proteins (Bcl-2 and caspase-3) were also respectively assessed by immunoblotting. The data showed that mice pretreated with NEC-1 reduced the amount of PI-positive cells from 12 to 48 h after TBI. Immunoblotting results showed that NEC-1 suppressed TBI-induced Beclin-1 and LC3-II activation which maintained p62 at high level. NEC-1 pretreatment also reversed TBI-induced Bcl-2 expression and caspase-3 activation, as well as the ratio of Beclin-1/Bcl-2. Both 3-MA and NEC-1 suppressed TBI-induced caspase-3 activation and LC3-II formation, Z-VAD only inhibited caspase-3 activation but increased LC3-II expression at 24 h post-TBI. All these results revealed that multiple cell death pathways participated in the development of TBI, and NEC-1 inhibited apoptosis and autophagy simultaneously. These coactions may further explain how can NEC-1 reduce TBI-induced tissue damage and functional deficits and reflect the interrelationship among necrosis, apoptosis and autophagy.
Department of Forensic Science and Laboratory of Neural Injury, Medical College of Soochow University, Suzhou 215123, Jiangsu, People's Republic of China.
Yao-Qi
YQ
Wang
Long
L
Wang
Ming-Yang
MY
Zhang
Tao
T
Wang
Hai-Jun
HJ
Bao
Wei-Li
WL
Liu
Ding-Kun
DK
Dai
Lu
L
Zhang
Pan
P
Chang
Wen-Wen
WW
Dong
Xi-Ping
XP
Chen
Lu-Yang
LY
Tao
D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001343:Autophagy / Q000187:drug effects*; D015153:Blotting, Western; D001930:Brain Injuries / Q000473:pathology*; D053148:Caspase 3 / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D004396:Coloring Agents; D004789:Enzyme Activation; D007093:Imidazoles / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D007211:Indoles / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D007276:Injections, Intraventricular; D008297:Male; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis / Q000235:genetics; D009842:Oligopeptides / Q000494:pharmacology; D011419:Propidium; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis
D004396:Coloring Agents; D007093:Imidazoles; D007211:Indoles; C480860:Map1lc3b protein, mouse; D008869:Microtubule-Associated Proteins; D009842:Oligopeptides; D019253:Proto-Oncogene Proteins c-bcl-2; C517629:benzyloxycarbonyl-valyl-alanyl-aspartic acid; C507699:necrostatin-1; D011419:Propidium; C025946:3-methyladenine; D053148:Caspase 3; D000225:Adenine