title: Activation of autophagy in photoreceptor necroptosis after experimental retinal detachment.
abstract: OBJECTIVE
  To investigate whether photoreceptor necroptosis induced by z-VAD-FMK (pan caspase inhibitor) was involved the activation of autophagy and whether Necrostatin-1, a specific necroptosis inhibitor, could inhibit this induction of autophagy after experimental retinal detachment.
  
  
  METHODS
  Experimental retinal detachment models were created in Sprague-Dawley rats by subretinal injection of sodium hyaluronate and subretinal injections of z-VAD-FMK, vehicle or z-VAD-FMK plus Necrostatin-1. Three days after retinal detachment, morphologic changes were observed by transmission electron microscopy. In other animals, retinas were subjected to immunoprecipitation and Western Blotting, then probed with anti-RIP1, phosphoserine, LC-3II or caspase 8 antibody.
  
  
  RESULTS
  It was proved by immunoprecipitation and western blotting, that photoreceptor necroptosis was mediated by caspase-8 inhibition and receptor interacting protein kinase (RIP1) phosphorylation activation. Transmission electron microscope and western blotting results indicated that photoreceptor necroptosis was involved the LC-3II and autophagosomes induction. We also discovered Necrostatin-1 could inhibit RIP1 phosphorylation and LC-3II induction.
  
  
  CONCLUSIONS
  These data firstly indicate photoreceptor necroptosis is associated with the activation of autophagy. Necrostatin-1 protects photoreceptors from necroptosis and autophagy by down-regulation of RIP1 phosphorylation and LC-3II.
authors.affiliation: Department of Ophthalmology, Anhui Provincial Hospital, Anhui Medical University, Hefei 230001, Anhui Province, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Dong
authors.affiliation: Department of Ophthalmology, Anhui Provincial Hospital, Anhui Medical University, Hefei 230001, Anhui Province, China.
authors.forename: Zi-Cheng
authors.initials: ZC
authors.lastname: Zhu
authors.affiliation: Department of Ophthalmology, Shanghai First People's Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200080, China ; Eye Research Institute of Shanghai Jiaotong University, Shanghai 200080, China.
authors.forename: Feng-Hua
authors.initials: FH
authors.lastname: Wang
authors.affiliation: Department of Ophthalmology, Anhui Provincial Hospital, Anhui Medical University, Hefei 230001, Anhui Province, China.
authors.forename: Gen-Jie
authors.initials: GJ
authors.lastname: Ke
authors.affiliation: Department of Morphology, Fudan University Shanghai Medical College, Shanghai 200080, China.
authors.forename: Zhang
authors.initials: Z
authors.lastname: Yu
authors.affiliation: Department of Ophthalmology, Shanghai First People's Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200080, China ; Shanghai Key Laboratory of Fundus Disease, Shanghai 200080, China.
authors.forename: Xun
authors.initials: X
authors.lastname: Xu
keywords: autophagy; necroptosis; retinal detachment