title: MLIF Alleviates SH-SY5Y Neuroblastoma Injury Induced by Oxygen-Glucose Deprivation by Targeting Eukaryotic Translation Elongation Factor 1A2.
abstract: Monocyte locomotion inhibitory factor (MLIF), a heat-stable pentapeptide, has been shown to exert potent anti-inflammatory effects in ischemic brain injury. In this study, we investigated the neuroprotective action of MLIF against oxygen-glucose deprivation (OGD)-induced injury in human neuroblastoma SH-SY5Y cells. MTT assay was used to assess cell viability, and flow cytometry assay and Hoechst staining were used to evaluate apoptosis. LDH assay was used to exam necrosis. The release of inflammatory cytokines was detected by ELISA. Levels of the apoptosis associated proteins were measured by western blot analysis. To identify the protein target of MLIF, pull-down assay and mass spectrometry were performed. We observed that MLIF enhanced cell survival and inhibited apoptosis and necrosis by inhibiting p-JNK, p53, c-caspase9 and c-caspase3 expression. In the microglia, OGD-induced secretion of inflammatory cytokines was markedly reduced in the presence of MLIF. Furthermore, we found that eukaryotic translation elongation factor 1A2 (eEF1A2) is a downstream target of MLIF. Knockdown eEF1A2 using short interfering RNA (siRNA) almost completely abrogated the anti-apoptotic effect of MLIF in SH-SY5Y cells subjected to OGD, with an associated decrease in cell survival and an increase in expression of p-JNK and p53. These results indicate that MLIF ameliorates OGD-induced SH-SY5Y neuroblastoma injury by inhibiting the p-JNK/p53 apoptotic signaling pathway via eEF1A2. Our findings suggest that eEF1A2 may be a new therapeutic target for ischemic brain injury.
authors.affiliation: Department of Pharmacology, College of Pharmacy, Second Military Medical University, Shanghai, China.
authors.forename: Qiuzhen
authors.initials: Q
authors.lastname: Zhu
authors.affiliation: Department of Pharmacology, College of Pharmacy, Second Military Medical University, Shanghai, China.
authors.forename: Yuefan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Pharmacology, College of Pharmacy, Second Military Medical University, Shanghai, China.
authors.forename: Yulan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: College of Pharmacy, Anhui University of Chinese Medicine, Hefei, Anhui, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Cheng
authors.affiliation: College of Pharmacy, Anhui University of Chinese Medicine, Hefei, Anhui, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.affiliation: College of Pharmacy, Anhui University of Chinese Medicine, Hefei, Anhui, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Pharmacology, College of Pharmacy, Second Military Medical University, Shanghai, China.
authors.forename: Yaocheng
authors.initials: Y
authors.lastname: Rui
authors.affiliation: Department of Pharmacology, College of Pharmacy, Second Military Medical University, Shanghai, China.
authors.forename: Tiejun
authors.initials: T
authors.lastname: Li
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology* / Q000627:therapeutic use; D017209:Apoptosis / Q000187:drug effects; D001930:Brain Injuries / Q000188:drug therapy* / Q000209:etiology / Q000473:pathology; D002545:Brain Ischemia / Q000150:complications / Q000188:drug therapy* / Q000473:pathology; D053148:Caspase 3 / Q000096:biosynthesis; D053453:Caspase 9 / Q000096:biosynthesis; D015687:Cell Hypoxia / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D016207:Cytokines / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors; D051379:Mice; D017628:Microglia / Q000473:pathology; D008954:Models, Biological; D058990:Molecular Targeted Therapy; D018696:Neuroprotective Agents / Q000494:pharmacology* / Q000627:therapeutic use; D009842:Oligopeptides / Q000494:pharmacology* / Q000627:therapeutic use; D020648:Peptide Elongation Factor 1 / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D020521:Stroke / Q000188:drug therapy / Q000473:pathology; D016159:Tumor Suppressor Protein p53 / Q000037:antagonists & inhibitors
chemical_list: D000893:Anti-Inflammatory Agents; D016207:Cytokines; C503234:EEF1A2 protein, human; D018696:Neuroprotective Agents; D009842:Oligopeptides; D020648:Peptide Elongation Factor 1; D034741:RNA, Small Interfering; D016159:Tumor Suppressor Protein p53; C430647:monocyte locomotion inhibitory factor; D048031:JNK Mitogen-Activated Protein Kinases; D053148:Caspase 3; D053453:Caspase 9; D005947:Glucose