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LncRNA CEBPA-AS1 Alleviates Cerebral Ischemia-Reperfusion Injury by Sponging miR-340-5p to Regulate APPL1/LKB1/AMPK Pathway
Xiankun Tu
Fujian Medical University Union Hospital
Corresponding Author
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BACKGROUND: The aim of this study was to research the mechanism of lncRNA CEBPA-AS1 in cerebral ischemia-reperfusion injury (CIRI).
METHODS: Middle cerebral artery occlusion (MCAO), MCAO/IR and OGD/R models were constructed. RNA immunoprecipitation (RIP) detected the binding of miR-340-5p to CEBPA-AS1. CEBPA-AS1 and miR-340-5p was knockdown or up-regulated. RT-PCR and western blot was processed to detect the expression of related genes and proteins. Brain tissue water content in each group was determined. Nissl staining in hippocampus of the brain and NeuN staining (green) assay was used to observe nerve damage and detect nerve cell survival, respectively. Cell viability was detected by MTT assay. Nerve cell survival was observed by immunofluorescence cytochemistry assay. LDH and MDA content were detected by kit. TUNEL was used to detect apoptosis. Luciferase reporter system was processed for verifying the binding sites. After overexpressing CEBPA-AS1 in PC-12 cells, RNA pulldown assay was performed. TTC assay was used to observe the general morphology of the brain.
RESULTS: Overexpression of CEBPA-AS1 attenuated MCAO/IR-induced nerve damage. Increased CEBPA-AS1 expression reduced neuronal apoptosis in MCAO/IR model. Knockdown of CEBPA-AS1 aggravated cell damage OGD/R cell model. CEBPA-AS1 increased APPL1 expression via negatively regulating miR-340-5p, and affected APPL1/LKB1/AMPK pathway. CEBPA-AS1 attenuated OGD/R-induced cell damage by reducing miR-340-5p levels.
CONCLUSIONS: LncRNA CEBPA-AS1 could alleviate cerebral ischemia-reperfusion injury by sponging miR-340-5p to regulate APPL1/LKB1/AMPK pathway.
Keywords
cerebral ischemia-reperfusion injury, CEBPA-AS1, miR-340-5p, APPL1/LKB1/AMPK pathway, MCAO/IR rat model
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Research
LncRNA CEBPA-AS1 Alleviates Cerebral Ischemia-Reperfusion Injury by Sponging miR-340-5p to Regulate APPL1/LKB1/AMPK Pathway
Xiankun Tu
Fujian Medical University Union Hospital
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 09 Jun, 2020
No community comments so far
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Neurobiology of Disease
License:
This work is licensed under a CC BY 4.0 License. Read Full License
BACKGROUND: The aim of this study was to research the mechanism of lncRNA CEBPA-AS1 in cerebral ischemia-reperfusion injury (CIRI).
METHODS: Middle cerebral artery occlusion (MCAO), MCAO/IR and OGD/R models were constructed. RNA immunoprecipitation (RIP) detected the binding of miR-340-5p to CEBPA-AS1. CEBPA-AS1 and miR-340-5p was knockdown or up-regulated. RT-PCR and western blot was processed to detect the expression of related genes and proteins. Brain tissue water content in each group was determined. Nissl staining in hippocampus of the brain and NeuN staining (green) assay was used to observe nerve damage and detect nerve cell survival, respectively. Cell viability was detected by MTT assay. Nerve cell survival was observed by immunofluorescence cytochemistry assay. LDH and MDA content were detected by kit. TUNEL was used to detect apoptosis. Luciferase reporter system was processed for verifying the binding sites. After overexpressing CEBPA-AS1 in PC-12 cells, RNA pulldown assay was performed. TTC assay was used to observe the general morphology of the brain.
RESULTS: Overexpression of CEBPA-AS1 attenuated MCAO/IR-induced nerve damage. Increased CEBPA-AS1 expression reduced neuronal apoptosis in MCAO/IR model. Knockdown of CEBPA-AS1 aggravated cell damage OGD/R cell model. CEBPA-AS1 increased APPL1 expression via negatively regulating miR-340-5p, and affected APPL1/LKB1/AMPK pathway. CEBPA-AS1 attenuated OGD/R-induced cell damage by reducing miR-340-5p levels.
CONCLUSIONS: LncRNA CEBPA-AS1 could alleviate cerebral ischemia-reperfusion injury by sponging miR-340-5p to regulate APPL1/LKB1/AMPK pathway.
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