ABSTRACT
Long noncoding RNAs (lncRNAs) have been validated to mediate the development of atherosclerosis (AS). In the present study, the molecular mechanisms and functions of lncRNA nuclear paraspeckle assembly transcript 1 (NEAT1) in the advancement of human aortic endothelial cells (HAECs) were investigated. The levels of lncRNANEAT1 and miR638 expression in clinical samples and cells were explored via quantitative reverse transcription polymerase chain reaction. Colony formation and CCK8 assays were performed to determine the proliferative capacity of cells, and the apoptotic capacity of cells was analyzed on the basis of apoptotic cell proportion and caspase3 activity. Then, the proportion of cells and correlations among phosphoglycerate kinase 1 (PGK1), NEAT1, and miR638 were determined through RNA immunoprecipitation and luciferase assays and bioinformatics analysis. Moreover, the expression levels of Ki67, proliferating cell nuclear antigen, PGK1, Bax, Bcl2, (p)mTOR, (p)AKT, and ßcatenin were analyzed via western blot analysis. In the serum of patients with AS and HAECs induced by oxidized lowdensity lipoprotein (oxLDL), the expression level of miR638 was decreased, whereas that of NEAT1 was increased. After oxLDL therapy, NEAT1 knockdown suppressed HAEC proliferation and stimulated HAEC apoptosis, which could be reversed by the miR638 inhibitor. NEAT1 inhibited miR638 expression through direct mutual action. The following mechanical investigations revealed that PGK1 was a miR638 target, whose expression was increased by NEAT1, a competing endogenous RNA of miR638. Additionally, the miR638 inhibitor contributed to proliferation and suppressed apoptosis through the activation of the AKT/mTOR signaling pathway in oxLDLinduced HAECs. NEAT1 adjusted the AKT/mTOR signaling pathway via miR638 in oxLDLinduced HAECs to accelerate their proliferation and impede their apoptosis. This result revealed that NEAT1 may be valuable in the treatment of AS.
Subject(s)
Atherosclerosis/genetics , Endothelial Cells/cytology , MicroRNAs/genetics , RNA, Long Noncoding/genetics , Atherosclerosis/metabolism , Cell Line , Cell Proliferation , Cell Survival , Endothelial Cells/chemistry , Endothelial Cells/drug effects , Female , Gene Knockdown Techniques , Humans , Lipoproteins, LDL/adverse effects , Male , Proto-Oncogene Proteins c-akt/metabolism , Signal Transduction , TOR Serine-Threonine Kinases/metabolismABSTRACT
Total flavones of Abelmoschus manihot L. Medic (TFA) is the major active component isolated from the traditional Chinese herb Abelmoschus manihot L. Medic. We investigated the protective effect of TFA against poststroke depression (PSD) injury in mice and its action mechanism. A mouse model of PSD was induced by middle cerebral artery occlusion (MACO) 30 min/reperfusion, followed by isolation feeding and chronic unpredictable mild stress for 2 weeks. Treatment groups received TFA at three different doses (160, 80, and 40 mg/kg, p.o.) or fluoxetine (Flu, 2.5 mg/kg, p.o.) daily for 24 days. Change in behavior, brain tissue malondialdehyde (MDA) levels, and the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were measured. The expression of brain-derived neurotrophic factor (BDNF) was detected by immunohistochemistry, and mRNA expression of BDNF and cAMP response element-binding protein (CREB) analyzed by reverse transcription-polymerase chain reaction (RT-PCR). Treatment with TFA (160, 80, and 40 mg/kg) significantly ameliorated mice escape-directed behavioral impairment induced by PSD, markedly reduced MDA levels, and increased the activity of SOD, GSH-Px close to normal levels. TFA administration also attenuated PSD-induced neuronal death/losses, upregulated expression of BDNF both at mRNA and protein levels, as well as CREB mRNA levels. TFA had a protective effect against PSD injury in mice. Cardioprotection involves the inhibition of lipid peroxidation and upregulation of BDNF-CREB levels in the hippocampus, which may also be important mechanism of its antidepressants. This potential protection makes TFA a promising therapeutic agent for the PSD.
Subject(s)
Abelmoschus/chemistry , Brain Injuries/drug therapy , Depressive Disorder/drug therapy , Drugs, Chinese Herbal/therapeutic use , Flavones/therapeutic use , Phytotherapy , Stroke/pathology , Animals , Behavior, Animal , Brain Injuries/metabolism , Brain-Derived Neurotrophic Factor/genetics , Brain-Derived Neurotrophic Factor/metabolism , Cyclic AMP Response Element-Binding Protein/genetics , Cyclic AMP Response Element-Binding Protein/metabolism , Depressive Disorder/metabolism , Glutathione Peroxidase/metabolism , Hippocampus/drug effects , Infarction, Middle Cerebral Artery/chemically induced , Infarction, Middle Cerebral Artery/complications , Lipid Peroxidation , Male , Malondialdehyde/metabolism , Mice , RNA, Messenger/genetics , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Stroke/etiology , Superoxide Dismutase/metabolismABSTRACT
Transfection of cDNA in 3'untranslated region of human nuclear factor for interleukin-6 (NF-IL6 3'UTR) induced tumor suppression in a human hepatoma cell line. cDNA array analysis was used to reveal changes in gene expression profile leading to tumor suppression The results indicate that this suppression was not due to activation of dsRNA-dependent protein kinase, nor to inactivation of oncogenes; rather, all the changes in expression of known genes, induced by NF-IL6 3'UTR cDNA may be ascribed to the suppression of cellular malignancy. Therefore, our results imply that this 3'untranslated region may have played role of a regulator of gene expression profile.
