title: Inhibition of calpain on oxygen glucose deprivation-induced RGC-5 necroptosis.
abstract: The purpose of this study was to investigate the effect of inhibition of calpain on retinal ganglion cell-5 (RGC-5) necroptosis following oxygen glucose deprivation (OGD). RGC-5 cells were cultured in Dulbecco's-modified essential medium and necroptosis was induced by 8-h OGD. PI staining and flow cytometry were performed to detect RGC-5 necrosis. The calpain expression was detected by Western blotting and immunofluorescence staining. The calpain activity was tested by activity detection kit. Flow cytometry was used to detect the effect of calpain on RGC-5 necroptosis following OGD with or without N-acetyl-leucyl-leucyl-norleucinal (ALLN) pre-treatment. Western blot was used to detect the protein level of truncated apoptosis inducing factor (tAIF) in RGC-5 cells following OGD. The results showed that there was an up-regulation of the calpain expression and activity following OGD. Upon adding ALLN, the calpain activity was inhibited and tAIF was reduced following OGD along with the decreased number of RGC-5 necroptosis. In conclusion, calpain was involved in OGD-induced RGC-5 necroptosis with the increased expression of its downstream molecule tAIF.
authors.affiliation: Department of Anatomy and Neurobiology, Central South University, Changsha, 410013, China.
authors.forename: Shuang
authors.initials: S
authors.lastname: Chen
authors.affiliation: Department of Forensic Science, School of Basic Medical Sciences, Central South University, Changsha, 410013, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Yan
authors.affiliation: Five-year Medicine Program, Grade 2013, Xiangya School of Medicine, Central South University, Changsha, 410013, China.
authors.forename: Hai-Xiao
authors.initials: HX
authors.lastname: Deng
authors.affiliation: Department of Forensic Science, School of Basic Medical Sciences, Central South University, Changsha, 410013, China.
authors.forename: Ling-Ling
authors.initials: LL
authors.lastname: Long
authors.affiliation: Department of Cardiology, People's Hospital of Hunan Province, Changsha, 410008, China.
authors.forename: Yong-Jun
authors.initials: YJ
authors.lastname: Hu
authors.affiliation: Department of Anatomy and Neurobiology, Central South University, Changsha, 410013, China.
authors.forename: Mi
authors.initials: M
authors.lastname: Wang
authors.affiliation: Jiangxi Research Institute of Ophthalmology and Visual Sciences, Affiliated Eye Hospital of Nanchang University, Nanchang, 330006, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Shang
authors.affiliation: Department of Anatomy and Neurobiology, Central South University, Changsha, 410013, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Chen
authors.affiliation: Department of Anatomy and Neurobiology, Central South University, Changsha, 410013, China.
authors.forename: Ju-Fang
authors.initials: JF
authors.lastname: Huang
authors.affiliation: Department of Anatomy and Neurobiology, Central South University, Changsha, 410013, China. xiongkun2001@163.com.
authors.forename: Kun
authors.initials: K
authors.lastname: Xiong
mesh_terms: D000818:Animals; D051033:Apoptosis Inducing Factor / Q000096:biosynthesis / Q000235:genetics*; D002154:Calpain / Q000096:biosynthesis / Q000235:genetics*; D005786:Gene Expression Regulation / Q000187:drug effects; D005947:Glucose / Q000378:metabolism; D006801:Humans; D007976:Leupeptins / Q000008:administration & dosage; D051379:Mice; D010100:Oxygen / Q000378:metabolism; D012165:Retinal Ganglion Cells / Q000378:metabolism* / Q000473:pathology; D015882:Retinal Necrosis Syndrome, Acute / Q000235:genetics* / Q000473:pathology
chemical_list: D051033:Apoptosis Inducing Factor; D007976:Leupeptins; C494830:AIFM1 protein, mouse; C063261:acetylleucyl-leucyl-norleucinal; D002154:Calpain; D005947:Glucose; D010100:Oxygen
keywords: N-acetyl-leucyl-leucyl-norleucinal; calpain; necroptosis; oxygen glucose deprivation; retinal ganglion cell-5; truncated apoptosis inducing factor