Inhibition of miR-128 Abates Aβ-Mediated Cytotoxicity by Targeting PPAR-γ via NF-κB Inactivation in Primary Mouse Cortical Neurons and Neuro2a Cells
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Purpose: Alzheimer’s disease (AD) is the sixth most common cause of death in the United States. MicroRNAs have been identifiedas vital players in neurodegenerative diseases, including AD. microRNA-128 (miR-128) has been shown to be dysregulated inAD. This study aimed to explore the roles and molecular mechanisms of miR-128 in AD progression. Materials and Methods: Expression patterns of miR-128 and peroxisome proliferator-activated receptor gamma (PPAR-γ) messengerRNA in clinical samples and cells were measured using RT-qPCR assay. PPAR-γ protein levels were determined by Westernblot assay. Cell viability was determined by MTT assay. Cell apoptotic rate was detected by flow cytometry via double-stainingof Annexin V-FITC/PI. Caspase 3 and NF-κB activity was determined by a Caspase 3 Activity Assay Kit or NF-κB p65 TranscriptionFactor Assay Kit, respectively. Bioinformatics prediction and luciferase reporter assay were used to investigate interactions betweenmiR-128 and PPAR-γ 3'UTR. Results: MiR-128 expression was upregulated and PPAR-γ expression was downregulated in plasma from AD patients andamyloid-β (Aβ)-treated primary mouse cortical neurons (MCN) and Neuro2a (N2a) cells. Inhibition of miR-128 decreased Aβ-mediated cytotoxicity through inactivation of NF-κB in MCN and N2a cells. Moreover, PPAR-γ was a target of miR-128. PPAR-γupregulation attenuated Aβ-mediated cytotoxicity by inactivating NF-κB in MCN and N2a cells. Furthermore, PPAR-γ downregulationwas able to abolish the effect of anti-miR-128 on cytotoxicity and NF-κB activity in MCN and N2a cells. Conclusion: MiR-128 inhibitor decreased Aβ-mediated cytotoxicity by upregulating PPAR-γ via inactivation of NF-κB in MCNand N2a cells, providing a new potential target in AD treatment.
Abstract
Purpose: Alzheimer’s disease (AD) is the sixth most common cause of death in the United States. MicroRNAs have been identifiedas vital players in neurodegenerative diseases, including AD. microRNA-128 (miR-128) has been shown to be dysregulated inAD. This study aimed to explore the roles and molecular mechanisms of miR-128 in AD progression. Materials and Methods: Expression patterns of miR-128 and peroxisome proliferator-activated receptor gamma (PPAR-γ) messengerRNA in clinical samples and cells were measured using RT-qPCR assay. PPAR-γ protein levels were determined by Westernblot assay. Cell viability was determined by MTT assay. Cell apoptotic rate was detected by flow cytometry via double-stainingof Annexin V-FITC/PI. Caspase 3 and NF-κB activity was determined by a Caspase 3 Activity Assay Kit or NF-κB p65 TranscriptionFactor Assay Kit, respectively. Bioinformatics prediction and luciferase reporter assay were used to investigate interactions betweenmiR-128 and PPAR-γ 3'UTR. Results: MiR-128 expression was upregulated and PPAR-γ expression was downregulated in plasma from AD patients andamyloid-β (Aβ)-treated primary mouse cortical neurons (MCN) and Neuro2a (N2a) cells. Inhibition of miR-128 decreased Aβ-mediated cytotoxicity through inactivation of NF-κB in MCN and N2a cells. Moreover, PPAR-γ was a target of miR-128. PPAR-γupregulation attenuated Aβ-mediated cytotoxicity by inactivating NF-κB in MCN and N2a cells. Furthermore, PPAR-γ downregulationwas able to abolish the effect of anti-miR-128 on cytotoxicity and NF-κB activity in MCN and N2a cells. Conclusion: MiR-128 inhibitor decreased Aβ-mediated cytotoxicity by upregulating PPAR-γ via inactivation of NF-κB in MCNand N2a cells, providing a new potential target in AD treatment.
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