Abstract
Objective: lncRNAs are recently thought to play a significant role in cellular homeostasis during pathological process of diseases by competing inhibiting miRNA function. The aim of present study was to assess the function of long non-coding RNA (lncRNA) MEG3 and its functional interaction with microRNA-181b in cerebral ischemic infarct of mice and hypoxia-induced neurons apoptosis.Methods: To address this question, we performed the experiments with in vivo middle cerebral artery occlusion (MCAO) mice model and in vitro oxygen-glucose deprivation (OGD)-cultured neuronal HT22 cell line. Relative expression of MEG3, miR-181b, and 12/15-LOX (lipoxygenase) mRNA was determined using quantitative RT-PCR. Western blot was used to evaluate 12/15-LOX protein expression. TUNEL assay was performed to assess cell apoptosis.Results: In both MCAO mice and OGD-cultured HT22 cell, ischemia, or hypoxia treatment results in a time-dependent increase in MEG3 and 12/15-LOX expression and decrease in miR-181b expression. Knockdown of MEG3 contributes to attenuation of hypoxia-induced apoptosis of HT22 cell. Also, expression level of MEG3 negatively correlated with miR-181b expression and positively correlated with 12/15-LOX expression. In contrary to MEG3, miR-181b overexpression attenuated hypoxia-induced HT22 cell apoptosis, as well as suppressed hypoxia-induced increase in 12/15-LOX expression. By luciferase reporter assay, we concluded that miR-181b directly binds to 12/15-LOX 3′-UTR, thereby negatively regulates 12/15-LOX expression.Conclusion: Our data suggested that long non-coding RNA MEG3 functions as a competing endogenous RNA for miR-181b to regulate 12/15-LOX expression in middle cerebral artery occlusion-induced ischemic infarct of brain nerve cells.
Highlights
In recent years, with the increasing incidence of cerebrovascular disease, the nerve injury caused by acute cerebral ischemia brings an increasingly heavy burden on individuals and society
We currently evaluated whether ischemia-responsive lncRNA interact with miR-181b, associated with genetic phenotype change of 12/415-LOX in cerebral ischemic mice
To examine aberrant genes expression in response to ischemia, mice middle cerebral artery occlusion model was established and infarction tissues were isolated for subsequent analysis of quantitative RT-PCR and western blot
Summary
With the increasing incidence of cerebrovascular disease, the nerve injury caused by acute cerebral ischemia brings an increasingly heavy burden on individuals and society This situation makes it a central issue to elucidate the molecular mechanisms of acute cerebral ischemia-related neuronal injury (Chen et al, 2014). According to encode project, ∼80% of the human genome is transcribed to RNA, of which ∼90% genome transcribes into non-coding RNA. These non-coding RNAs are increasing recognized and defined as endogenous regulatory RNA involved in maintenance of cellular homeostasis during growth, development and pathogenesis disease. LncRNA is recently found to be a competing endogenous RNA for miRNA and these interactions have been found in various pathological mechanisms (Tay et al, 2014)
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