Effect of CircOGDH on Oxygen-Glucose Deprivation/Reoxygenation-Induced Microglial Cell Injury through Modulation of the miR-195-5p/PGAM5 Axis
Objective:To investigate the effect of circular RNA OGDH(CircOGDH)on microglia(MG)cell damage induced by oxygen-glucose deprivation/reoxygenation(OGD/R)by modulating the miR-195-5p/phosphoglycerate mutase 5(PGAM5)axis.Methods:HMC3 cells cultured in vitro were separated into:NC group(normal culture),OGD/R,si-NC group,si-CircOGDH group,mimic NC group,miR-195-5p mimic group,si-CircOGDH+inhibitor NC group,and si-CircOGDH+miR-195-5p inhibitor group.Except for the NC group,the remaining groups constructed OGD/R models before transfecting the corresponding sub-stances.QRT-PCR method was applied to determine the expression levels of CircOGDH,miR-195-5p,and PGAM5 mRNA.The levels of cell proliferation,apoptosis,reactive oxygen species(ROS),and inflammatory factors were measured in each group.Western blot was applied to detect the expression of Cleaved caspase-3 and Bax.Dual luciferase gene experiment was used to detect the targeting relationship between CircOGDH and miR-195-5p,and between PGAM5 and miR-195-5p.Results:Compared with the NC group,the expres-sion of miR-195-5p and OD450 value were reduced in the OGD/R group,the mRNA expression of CircOG-DH and PGAM5,apoptosis rate,ROS,and inflammatory cytokine levels,and the expression of Cleaved caspase-3 and Bax proteins were all elevated(P<0.05).Compared with the OGD/R group and the si-NC group,the expression of miR-195-5p and OD450 value were elevated in the si-CircOGDH group,the mRNA expression of CircOGDH and PGAM5,apoptosis rate,ROS and inflammatory cytokine levels,and the expres-sion of Cleaved caspase-3 and Bax proteins were all reduced(P<0.05).Compared with the OGD/R group and the mimic NC group,the miR-195-5p mimic group showed no significant difference in CircOGDH(P>0.05),while the expression of miR-195-5p and OD450 values were increased,the expression of PGAM5 mRNA,apoptosis rate,ROS and inflammatory cytokine levels,and the expression of Cleaved caspase-3 and Bax protein were all reduced(P<0.05).Compared with the si-CircOGDH+inhibitor NC group,the CircOG-DH of the si-CircOGDH+miR-195-5p inhibitor group was not significantly different(P>0.05),the expres-sion of miR-195-5p and OD450 values were decreased,the expression of PGAM5 mRNA,apoptosis rate,ROS and inflammatory cytokine levels,and the expression of Cleaved caspase-3 and Bax protein were all ele-vated(P<0.05).A dual luciferase gene experiment showed that miR-195-5p had a targeting relationship with CircOGDH and PGAM5.Conclusion:Silencing CircOGDH can promote the proliferation of HMC3 cells induced by OGD/R,inhibit their inflammatory response and apoptosis,and may be related to the regulation of miR-195-5p/PGAM5 axis.
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