MiR-188-5p Targets SENP3 To Regulate Oxygen-glucose Deprivation/reoxygenation Injury In N2a Cells

Objective: To investigate the regulation of miR-188-5p targeting small ubiquitin-like modified protein-specific protease 3(SENP3)on oxygen-glucose deprivation/reoxygenation(OGD/R)injury in mouse neuroblastoma(N2a)cells and to explore its mechanism.Methods: N2 a cells were cultured to establish OGD/R model and randomly divided into five groups: control group(group C),group OGD/R,group OGD/R + NC,OGD/R + miR-188-5p mimics group(group OGD/R + mimics),and OGD/R + miR-188-5p inhibitor group(group OGD/R + inhibitor).Cells in group C were routinely cultured,and OGD/R + NC group,OGD/R + mimics group,and OGD/R + inhibitor group were transfected with reagent miR-188-5p negative control miR,mimics and inhibitor,respectively.The OGD/R model was subsequently established.Indicators:(1)A dual-luciferin experimental report was used to validate the targeting relationship of miR-188-5p and SENP3;(2)Detection at 24 hours after reoxygenation:(1)CCK-8 assay was used to detect the changes of cell viability in the five groups;(2)LDH leakage was used to assess the degree of cell damage;(3)Hoechst33258 staining was used to observe apoptosis;(4)Caspase-3 activity assay was used to assess the degree of apoptosis;(5)The expression levels of miR-188-5p and SENP3 m RNA were determined by q RT-PCR;(6)The expression of SENP3 in cells was measured by Western Blot.Results:(1)The dual-luciferin assay report demonstrated that SENP3 is a target gene of miR-188-5p,and the 3’UTR region of SENP3 m RNA contains a binding site for miR-188-5p.(2)Compared with group C,the cell viability of the other four groups were decreased(P<0.05),LDH leakage was increased(P<0.05),Hoechst 33258 staining showed that the number of apoptotic cells was increased,caspase-3 activity was increased(P<0.05),SENP3 m RNA and protein expression levels were up-regulated(P<0.05),miR-188-5p expression was increased in the OGD/R + mimics group(P<0.05),and decreased in the other three groups(P<0.05)(3)Compared with the group OGD/R,the cell viability of the group OGD/R+mimics increased(P<0.05),LDH leakage was decreased(P<0.05).Hoechst33258 staining showed that the number of apoptotic cells was decreased,the activity of caspase-3 was decreased(P<0.05),SENP3 m RNA and protein expression levels were decreased(P<0.05),miR-188-5p expression was increased(P<0.05).Compared with the group OGD/R,the cell viability of the group OGD/R+inhibitor was decreased(P<0.05),LDH leakage was increased(P<0.05),Hoechst33258 staining showed that the number of apoptotic cells was increased,caspase-3 activity was increased(P<0.05),SENP3 m RNA and protein expression levels were up-regulated(P<0.05),miR-188-5p expression was decreased(P<0.05).There was no significant difference between the group OGD/R+NC and the group OGD/R(P<0.05).Conclusions: miR-188-5p negatively regulates SENP3 to attenuate OGD/R injury in N2 a cells.