title: RIP3 induces ischemic neuronal DNA degradation and programmed necrosis in rat via AIF.
abstract: We have reported that nuclear translocation of Receptor-interacting protein 3 (RIP3) involves in neuronal programmed necrosis after 20-min global cerebral ischemia/reperfusion (I/R) injury. Herein, the underlying mechanisms and the nuclear role of RIP3 were investigated further. The necroptosis inhibitor necrostatin-1 (Nec-1), the autophagy inhibitor 3-methyladenine (3-MA), and the caspase-3 inhibitor acetyl-L-aspartyl-L-methionyl-L-glutaminyl-L-aspart-1-al (Ac-DMQD-CHO) were administered intracerebroventricularly 1 h before ischemia. Protein expression, location and interaction was determined by western blot, immunofluorescence or immunoprecipitation. Most CA1 neuronal death induced by 20-min global cerebral I/R injury was TUNEL-positive. Neuronal death and rat mortality rates were greatly inhibited by Nec-1 and 3-MA pre-treatment, but not by Ac-DMQD-CHO. And no activation of caspase-3 was detected after I/R injury. Caspase-8 was expressed richly in GFAP-positive astrocytes and Iba-1-positive microglia, but was not detected in Neun-positive neurons. The nuclear translocation and co-localization of RIP3 and AIF, and their interaction were detected after I/R injury. These processes were inhibited by Nec-1 and 3-MA pre-treatment, but not by Ac-DMQD-CHO. The formation of an RIP3-AIF complex and its nuclear translocation are critical to ischemic neuronal DNA degradation and programmed necrosis. Neurons are more likely to enter the programmed necrosis signal pathway for the loss of caspase-8 suppression.
authors.affiliation: Department of Neurology, Brain Medical Centre, First Affiliated Hospital, Zhejiang University School of Medicine, 89 Qingchun Road, Hangzhou 310003, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Department of Neurology, First Affiliated Hospital, Anhui Medical University, 218 Jixi Road, Hefei 230022, China.
authors.forename: Jingye
authors.initials: J
authors.lastname: Wang
authors.affiliation: Facility for Biochemistry and Molecular medicine Core Facilities, Zhejiang University School of Medicine, 866 Yuhangtang Road, Hangzhou 310058, China.
authors.forename: Xinghui
authors.initials: X
authors.lastname: Song
authors.affiliation: Department of Neurology, First Affiliated Hospital, Anhui Medical University, 218 Jixi Road, Hefei 230022, China.
authors.forename: Lindi
authors.initials: L
authors.lastname: Qu
authors.affiliation: Department of Neurology, Brain Medical Centre, First Affiliated Hospital, Zhejiang University School of Medicine, 89 Qingchun Road, Hangzhou 310003, China.
authors.forename: Ruili
authors.initials: R
authors.lastname: Wei
authors.affiliation: Department of Neurology, Brain Medical Centre, First Affiliated Hospital, Zhejiang University School of Medicine, 89 Qingchun Road, Hangzhou 310003, China.
authors.forename: Fangping
authors.initials: F
authors.lastname: He
authors.affiliation: Department of Neurology, First Affiliated Hospital, Anhui Medical University, 218 Jixi Road, Hefei 230022, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Wang
authors.affiliation: Department of Neurology, Brain Medical Centre, First Affiliated Hospital, Zhejiang University School of Medicine, 89 Qingchun Road, Hangzhou 310003, China.
authors.forename: Benyan
authors.initials: B
authors.lastname: Luo
mesh_terms: D000225:Adenine / Q000008:administration & dosage / Q000031:analogs & derivatives; D000818:Animals; D051033:Apoptosis Inducing Factor / Q000378:metabolism*; D002545:Brain Ischemia / Q000235:genetics / Q000378:metabolism / Q000401:mortality* / Q000473:pathology; D002467:Cell Nucleus / Q000378:metabolism; D053836:DNA Degradation, Necrotic; D004195:Disease Models, Animal; D007093:Imidazoles / Q000008:administration & dosage; D007211:Indoles / Q000008:administration & dosage; D008297:Male; D009474:Neurons / Q000378:metabolism / Q000473:pathology*; D009842:Oligopeptides / Q000008:administration & dosage; D051381:Rats; D017207:Rats, Sprague-Dawley; D053422:Receptor-Interacting Protein Serine-Threonine Kinases / Q000378:metabolism*; D015427:Reperfusion Injury / Q000235:genetics / Q000378:metabolism / Q000401:mortality* / Q000473:pathology
chemical_list: C494831:Aifm1 protein, rat; D051033:Apoptosis Inducing Factor; D007093:Imidazoles; D007211:Indoles; D009842:Oligopeptides; C110922:acetyl-aspartyl-methionyl-glutaminyl-aspartyl-aldehyde; C507699:necrostatin-1; C025946:3-methyladenine; D053422:Receptor-Interacting Protein Serine-Threonine Kinases; C582791:Ripk3 protein, rat; D000225:Adenine