title: Micro-RNA-30a regulates ischemia-induced cell death by targeting heat shock protein HSPA5 in primary cultured cortical neurons and mouse brain after stroke.
abstract: Micro-RNAs (miRs) have emerged as key gene regulators in many diseases, including stroke. We recently reported that miR-30a protects N2A cells against ischemic injury, in part through enhancing beclin 1-mediated autophagy. The present study explores further the involvement of miR-30a in ischemia-induced apoptosis and its possible mechanisms in primary cortical neurons and stroked mouse brain. We demonstrate that miR-30a level is significantly decreased in cortical neurons after 1-hr oxygen-glucose deprivation (OGD)/24-hr reoxygenation. Overexpression of miR-30a aggravated the OGD-induced neuronal cell death, whereas inhibition of miR-30a attenuated necrosis and apoptosis as determined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-di-phenyl-2H-tetrazolium bromide, lactate dehydrogenase, TUNEL, and cleaved caspase-3. The amount of HSPA5 protein, which is predicted to be a putative target of miR-30a by TargetScan, could be reduced by pre-miR-30a, whereas it was increased by anti-miR-30a. Furthermore, the luciferase reporter assay confirmed that miR-30a directly binds to the predicted 3'-UTR target sites of the hspa5 gene. The cell injury regulated by miR-30a in OGD-treated cells could be aggravated by HSPA5 siRNA. We also observed an interaction of HSPA5 and caspase-12 by coimmunoprecipitation and speculate that HSPA5 might be involved in endoplasmic reticulum stress-induced apoptosis. In vivo, reduced miR-30a increased the HSPA5 level and attenuated ischemic brain infarction in focal ischemia-stroked mice. Downregulation of miR-30a could prevent neural ischemic injury through upregulating HSPA5 protein expression, and decreased ER stress-induced apoptosis might be one of the mechanisms underlying HSPA5-mediated neuroprotection.
authors: Department of Neurobiology and Center of Stroke, Beijing Institute for Brain Disorders, Capital Medical University, Beijing, People's Republic of China.|Central Laboratory, Liaoning Medical University, Jinzhou, People's Republic of China.
authors: Peng
authors: P
authors: Wang
authors: Department of Anatomy, Capital Medical University, Beijing, People's Republic of China.
authors: Nan
authors: N
authors: Zhang
authors: Central Laboratory, Liaoning Medical University, Jinzhou, People's Republic of China.
authors: Jia
authors: J
authors: Liang
authors: Department of Neurobiology and Center of Stroke, Beijing Institute for Brain Disorders, Capital Medical University, Beijing, People's Republic of China.
authors: Jiefei
authors: J
authors: Li
authors: Department of Neurobiology and Center of Stroke, Beijing Institute for Brain Disorders, Capital Medical University, Beijing, People's Republic of China.
authors: Song
authors: S
authors: Han
authors: Department of Neurobiology and Center of Stroke, Beijing Institute for Brain Disorders, Capital Medical University, Beijing, People's Republic of China.
authors: Junfa
authors: J
authors: Li
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics*; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism / Q000473:pathology*; D002545:Brain Ischemia / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D004195:Disease Models, Animal; D000091342:Endoplasmic Reticulum Chaperone BiP; D005786:Gene Expression Regulation / Q000235:genetics; D006360:Heat-Shock Proteins / Q000378:metabolism*; D047468:Immunoprecipitation; D020287:In Situ Nick-End Labeling; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000502:physiology*; D009474:Neurons / Q000378:metabolism / Q000473:pathology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020521:Stroke / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D014161:Transduction, Genetic
chemical_list: D000091342:Endoplasmic Reticulum Chaperone BiP; D006360:Heat-Shock Proteins; C000717248:Hspa5 protein, mouse; D035683:MicroRNAs; C534715:Mirn30d microRNA, mouse
keywords: HSPA5; RRID:AB_10622025; RRID:AB_2069200; RRID:AB_331439; RRID:AB_476697; RRID:AB_776174; RRID:AB_823575; RRID:SciRes_000129; apoptosis; ischemic stroke; miRNA-30a