MicroRNA-219 alleviates glutamate-induced neurotoxicity in cultured hippocampal neurons by targeting calmodulin-dependent protein kinase II gamma
| Autor: | Wen-jie Yang, Qun Cai, Jian-Feng Yi, Feng Xu, Ting Wang, Hai-Hua Fan |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
caspase-3 miR-219 calmodulin-dependent protein kinase II γ hippocampal neurons Excitotoxicity Caspase 3 glutamate medicine.disease_cause Neuroprotection lcsh:RC346-429 03 medical and health sciences 0302 clinical medicine luciferase reporter gene system Developmental Neuroscience medicine MTT assay Viability assay nerve regeneration lcsh:Neurology. Diseases of the nervous system septic encephalopathy Chemistry Neurotoxicity Glutamate receptor apoptosis medicine.disease brain injury Cell biology 030104 developmental biology Apoptosis neuroprotection neural regeneration excitotoxicity 030217 neurology & neurosurgery Research Article |
| Zdroj: | Neural Regeneration Research Neural Regeneration Research, Vol 13, Iss 7, Pp 1216-1224 (2018) |
| ISSN: | 1876-7958 1673-5374 |
| Popis: | Septic encephalopathy is a frequent complication of sepsis, but there are few studies examining the role of microRNAs (miRs) in its pathogenesis. In this study, a miR-219 mimic was transfected into rat hippocampal neurons to model miR-219 overexpression. A protective effect of miR-219 was observed for glutamate-induced neurotoxicity of rat hippocampal neurons, and an underlying mechanism involving calmodulin-dependent protein kinase II γ (CaMKIIγ) was demonstrated. miR-219 and CaMKIIγ mRNA expression induced by glutamate in hippocampal neurons was determined by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR). After neurons were transfected with miR-219 mimic, effects on cell viability and apoptosis were measured by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry. In addition, a luciferase reporter gene system was used to confirm CaMKIIγ as a target gene of miR-219. Western blot assay and rescue experiments were also utilized to detect CaMKIIγ expression and further verify that miR-219 in hippocampal neurons exerted its effect through regulation of CaMKIIγ. MTT assay and qRT-PCR results revealed obvious decreases in cell viability and miR-219 expression after glutamate stimulation, while CaMKIIγ mRNA expression was increased. MTT, flow cytometry, and caspase-3 activity assays showed that miR-219 overexpression could elevate glutamate-induced cell viability, and reduce cell apoptosis and caspase-3 activity. Moreover, luciferase CaMKIIγ-reporter activity was remarkably decreased by co-transfection with miR-219 mimic, and the results of a rescue experiment showed that CaMKIIγ overexpression could reverse the biological effects of miR-219. Collectively, these findings verify that miR-219 expression was decreased in glutamate-induced neurons, CaMKIIγ was a target gene of miR-219, and miR-219 alleviated glutamate-induced neuronal excitotoxicity by negatively controlling CaMKIIγ expression. |
| Databáze: | OpenAIRE |
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