| Objective To explore the effect of long non coding RNA nuclear enriched abundant transcript 1 (LncRNA NEAT1) on sepsis-related brain injury by regulating the microRNA-34a-5p (miR-34a-5p)/lactate dehydrogenase A (LDHA) pathway. Methods A sepsis mouse model was established and classified into model group, si-NC group, si-NEAT1 group, anti-miR-NC group, anti-miR-34a-5p group, si-NEAT1+anti-miR-NC group, si-NEAT1+anti-miR-34a-5p group. The unmodeled mice were used as the control group. The neurobehavioral score was used to assess the degree of brain injury in mice. HE staining was performed to observe the morphological structure of mouse brain tissue. TUNEL staining was used to observe neuronal apoptosis in mouse brain tissue. The mouse hippocampal neuron cell line HT22 was classified into Control group, LPS group, LPS+si-NC group, LPS+si-NEAT1 group, LPS+si-NEAT1+anti-miR-NC group, and LPS+ si-NEAT1+anti-miR-34a-5p group. CCK-8 method and flow cytometry were used to evaluate cell proliferation and apoptosis, respectively. QRT-PCR was performed to detect the NEAT1, miR-34a-5p, and LDHA in tissues and cells. Western blot was performed to detect LDHA protein in tissues and cells. Dual luciferase assay was used to validate the targeting relationship between NEAT1 and miR-34a-5p, and between miR-34a-5p and LDHA. Results For the control group, the model group showed conspicuous neuronal injury in brain tissue, decreased neurobehavioral score and miR-34a-5p, increased NEAT1 and LDHA mRNAs and proteins, and neuronal apoptosis index (P<0.05). For the siNC group, the si-NEAT1 group showed conspicuous improved the pathological morphology of brain tissue, increased neurobehavioral score and miR-34a-5p, and decreased NEAT1 and LDHA mRNAs and proteins, and neuronal apoptosis index (P<0.05). For the anti-miR-NC group, the anti-miR-34a-5p group showed aggravated pathological damage of brain tissue in mice, decreased neurobehavioral score and miR-34a- 5p, and increased NEAT1 and LDHA mRNAs and proteins, and neuronal apoptosis index (P<0.05). For the Control group, the LPS group showed an increase in NEAT1 and LDHA mRNAs and proteins, and cell apoptosis rate, and a decrease in miR-34a-5p and OD value (P<0.05). For the LPS+si-NC group, the LPS+si-NEAT1 group showed an increase in miR-34a-5p and OD value, and a decrease in NEAT1 and LDHA mRNAs and proteins, and cell apoptosis rate (P<0.05). Inhibition of miR-34a-5p could weaken the improvement of NEAT1 downregulation on brain injury and LPS-induced HT22 cell damage in septic mice (P<0.05). Conclusion LncRNA NEAT1 may alleviate sepsisrelated brain injury by regulating the miR-34a-5p/LDHA pathway. |
