Effect of miR-182-5p on Neuron Damage Induced by Oxygen Glucose Deprivation/Reoxygenation by Targeting XBP1
MENG Zhimin1*, LIU Pin2, NI Xiujuan1, WANG Li1, QIAO Lina1
The article to investigate the effect of miR-182-5p on neuron damage induced by OGD/R (oxygen glucose deprivation/reoxygenation) by targeting endoplasmic reticulum stress-related factor XBP1 (X-box binding protein 1). PC-12 (rat nerve cells) were selected as the study subjects and randomly assigned into Control group, OGD/R group, miR-NC group (transfected with miR-NC), miR-182-5p mimics group (transfected with miR-182-5p mimics), miR-182-5p mimics+pcDNA-NC group (transfected with miR-182-5p mimics+pcDNANC), and miR-182-5p mimics+pcDNA-XBP1 group (transfected with miR-182-5p mimics+pcDNA-XBP1). Except for the control group, all other groups were induced with OGD/R. QRT-PCR was used to detect the mRNA expression of miR-182-5p and XBP1 in cells. CCK8 was used to detect cell proliferation. Flow cytometry was used to detect cell apoptosis rate. ELISA method for detecting the levels of SOD, MDA, IL-6, and TNF-α in cells. Western blot was used to detect the expression of PCNA, caspase-9, Bax, and XBP1 proteins in cells. Moreover, dual luciferase assay was used to detect the interaction between miR-182-5p and XBP1. The results show that, compared with the Control group, the OGD/R group showed a decrease in miR-182-5p expression, survival rate, clone number, SOD level, and PCNA protein expression (P<0.05), and an increase in apoptosis rate, MDA, IL-6, TNF-α levels, Bax, caspase-9, and XBP1 protein expression (P<0.05). Compared with the OGD/R group and miR-NC group, the miR-182-5p mimics group showed an increase in miR-182-5p expression, survival rate, clone number, SOD level, and PCNA protein expression (P<0.05), and a decrease in apoptosis rate, MDA, IL-6, TNF-α levels, Bax, caspase-9, and XBP1 protein expression (P<0.05). Compared with the miR- 182-5p mimics group and the miR-182-5p mimics+pcDNA-NC group, the miR-182-5p mimics+pcDNA-XBP1 group showed a decrease in survival rate, clone number, SOD level, and PCNA protein expression (P<0.05), and an increase in apoptosis rate, MDA, IL-6, TNF-α levels, Bax, caspase-9, and XBP1 protein expression (P<0.05). Compared with the miR-NC+XBP1-WT group, the miR-182-5p mimics+XBP1-WT group showed a obvious decrease in dual luciferase activity (P<0.05). In short, miR-182-5p may inhibit OGD/R-induced neuron damage by inhibiting XBP1 expression.
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