title: EIF2alpha and caspase-12 activation are involved in oxygen-glucose-serum deprivation/restoration-induced apoptosis of spinal cord astrocytes.
abstract: Astrocytes play an important role in protecting neurons during ischemia and reperfusion in the central nervous system. Although many studies have shown that oxygen-glucose deprivation (OGD) can induce astrocyte apoptosis, the role of PERK/eIF2 alpha/ATF4 integrated stress response (ISR) in astrocyte apoptosis mediated by oxygen-glucose-serum deprivation (OGSD)/restoration remains uncertain. Astrocytes were subjected to a combination of oxygen, glucose, and serum deprivation for 8h followed by restoration. Hoechst 33342 staining was performed to quantify apoptotic astrocytes and cell viability was assessed with Cell Counting Kit-8 (CCK8). Immunocytochemical analysis and Western blotting for some related molecules, including pancreatic ER stress kinase (PERK), p-PERK, eukaryotic initiation factor 2 alpha (eIF2 alpha), p-eIF2 alpha, activating transcription factor 4 (ATF4), caspase-12, were examined. Caspase activation and apoptosis were detected in neonatal rat astrocytes from spinal cord subjected to OGSD/restoration. We also observed an increase in cytoplasmic staining of p-eIF2 alpha in astrocytes (8h OGSD/15 min restoration) compared with that of non-treated cells. In addition, we found the sequential activation of PERK, eIF2 alpha, and ATF4 during OGSD/restoration by Western blotting. These results indicate that both the PERK/eIF2 alpha/ATF4 ISR and activation of caspase-12 may be involved in apoptosis of spinal cord astrocytes induced by OGSD/restoration.
authors.affiliation: Department of Orthopedics, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing 210029, China.
authors.forename: Ailiang
authors.initials: A
authors.lastname: Zhang
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.forename: Peng
authors.initials: P
authors.lastname: Sun
authors.forename: Changjiang
authors.initials: C
authors.lastname: Yao
authors.forename: Changhui
authors.initials: C
authors.lastname: Su
authors.forename: Tao
authors.initials: T
authors.lastname: Sui
authors.forename: Hua
authors.initials: H
authors.lastname: Huang
authors.forename: Xiaojian
authors.initials: X
authors.lastname: Cao
authors.forename: Yingbin
authors.initials: Y
authors.lastname: Ge
mesh_terms: D051701:Activating Transcription Factor 4 / Q000502:physiology; D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis; D001253:Astrocytes / Q000166:cytology* / Q000378:metabolism; D053454:Caspase 12 / Q000502:physiology*; D002470:Cell Survival; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D004789:Enzyme Activation; D015852:Eukaryotic Initiation Factor-2 / Q000502:physiology*; D005947:Glucose / Q000172:deficiency*; D010100:Oxygen / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013116:Spinal Cord / Q000166:cytology* / Q000378:metabolism; D019892:eIF-2 Kinase / Q000502:physiology
chemical_list: D051701:Activating Transcription Factor 4; D053454:Caspase 12; D016895:Culture Media, Serum-Free; D015852:Eukaryotic Initiation Factor-2; D005947:Glucose; D010100:Oxygen; D019892:eIF-2 Kinase; C497110:Atf4 protein, rat; C506443:Casp12 protein, rat