Objective To investigate the effect and mechanism of long non-coding RNA (lncRNA) nuclear-enriched abundant transcript 1(NEAT1)on hippocampal neuron apoptosis in epilepsy cell model, in order to provide evidence for NEAT1 being the new target of epilepsy treatment. Methods The rat hippocampal neuron cells were cultured in vitro and induced without magnesium to prepare epileptic hippocampal neuron models.The experiment included control group (normal extracellular fluid), model group (magnesium-free extracellular fluid), transfection control group (transfection non-specific siRNA+magnesium-free extracellular fluid) and transfection group(transfected with NEAT1 specific siRNA+ magnesium-free extracellular fluid).The expression changes of NEAT1 and miR-29b-3p were detected by real-time fluorescent quantitative PCR (qPCR).The dual luciferase reporter gene experiment detected whether NEAT1 targets miR-29b-3p, and interleukins-1(IL-1), interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) content were tested by enzyme-linked immunosorbent assay(ELISA) .AnnexinV-FITC/PI double staining method was used to detect hippocampal neuronal cell apoptosis, protein Western blot was used to detect the expression levels of B cell lymphoma/lewkmia-2 (Bcl-2), Bcl-2 Associated X Protein (Bax), toll-like receptor-4(TLR4) and nuclear factor-κB(NF-κB) in cells of each group. Results Compared with the control group, the apoptosis rate of hippocampal neurons and the expression levels of NEAT1, Bax, TLR4 and NF-κB in the model group increased (F=50.980, 73.668, 65.635, 13.203, 10.292, P<0.05), the contents of IL-1, IL-6 and TNF-α increased (F=33.107, 33.857, 51.129, P<0.05), the expression levels of miR-29b-3p and Bcl-2 decreased (F=145.023, 67.655, P<0.05).While inhibiting the expression of NEAT1 can reduce neuronal apoptosis, inhibit the expression of Bax, TLR4 and NF-κB, promote the expression of miR-29b-3p and Bcl-2, and reduce the secretion of IL-1, IL-6 and TNF-α.The dual luciferase reporter gene experiment confirmed the targeting relationship between NEAT1 and miR-29b-3p. Conclusion lncRNA NEAT1 targets to down-regulate the expression of miR-29b-3p and block the TLR4/NF-κB signaling pathway to inhibit the apoptosis of hippocampal neurons in epilepsy models.
Key words
lncRNA NEAT1 /
miR-29b-3p /
epilepsy /
hippocampal neuron apoptosis
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