In vitro experimental study of hypoxia-inducible factor-1 α in inhibiting microglial Pyroptosis in acute ischemic stroke by regulating MDA and SOD
Objective To explore the regulatory effect and mechanism of hypoxia-inducing factor-1α(HIF-1α)regulating malondialdehyde(MDA)and superoxide dismutase(SOD)on pyroptosis in microglial cells(BV2)in acute ischemic stroke.Methods The acute phase of oxygen and glucose deprivation/re-oxygenation(OGD/R)model of microglia was constructed,and the CCK-8 method was applied to determine the optimal intervention time of the model.HIF-1 α stabilizer GF-4592 overexpression and HIF-1 α small molecule interfering RNA(HIF-1 α-siRNA)suppressed expression and divided into blank group(group A),OGD/R model group(group B),OGD/R+FG-4592 intervention group(group C),OGD/R+siRNA negative control group(group D),and OGD/R+HIF-1α-siRNA group(group E).Cell proliferation in five groups was measured by CCK-8,and pyroptosis was determined by flow cytometry.MDA,SOD,IL-18 and IL-1β were measured by ELISA.Western blot Protein expression levels of HIF-1α,GSDMD-D,GSDMD-N,cle-Caspase-1and NLRP3 were measured in different groups.Results Determined 6 h reoxygenation was the optimal intervention time.Compared with group B and D,group C showed increased proliferation,decreased pyroptosis,MAD,SOD,IL-18,IL-1β,and decreased expression levels of GSDMD-D,GSDMD-N,clean-Caspase-1 and NLRP3 targets(P＜0.05),while group E decreased proliferation,increased cell pyroptosis and increased MAD,SOD,IL-18,IL-1β,GSDMD-D,GSDMD-N,clean-Caspase-1 and NLRP3 targets(P＜0.05).Conclusion The HIF-1 α factor can effectively regulate microglia pyroptosis.HIF-1α may inhibit microglia pyroptosis in acute ischemic stroke through the regulation of MDA and SOD.
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