Mechanism of Hydrogen Sulfide in Improving Hypoxia/Reoxygenation Injury in Rat Alveolar Epithelial Cell Type Ⅱ
This study aims to investigate the effect of H2S(hydrogen sulfide)preconditioning on H/R(hy-poxia/reoxygenation)injury in rat AECⅡ(alveolar epithelial cell type Ⅱ)and the underlying mechanism.The cells were identified by immunofluorescence.Detection of each cell activity,SOD(superoxide dismutase)activity,MDA(malondialdehyde)and lactate content were measured.The levels of IL-1β and IL-18 were detected by ELISA.The protein levels of aerobic glycolysis markers HK2 and PKM2 and pyroptosis markers NLRP3,GSDMD-N,cleaved-Caspasel,cleaved-IL-1β,and cleaved-IL-18 were detected by Western blot.qRT-PCR was used to detect the mRNA levels of HK2,PKM2,and NLRP3.Immunofluorescence was used to observe the expression of HK2 and NLRP3,and to evaluate the expression of aerobic glycolysis and pyroptosis indicators in H/R and the effect of H2S on H/R injury.In addition,to further investigate the link between aerobic glycolysis and pyroptosis,the HK2 inhibitor 2-DG(2-deoxyglucose)was used and its expression was detected by the laboratory methods described above.The results showed that H/R could activate aerobic glycolysis and pyroptosis.H2S treatment down-regulated the expression of aerobic glycolysis and pyroptosis indicators and ameliorated H/R-induced cell injury.After using 2-DG,pyroptosis indicators expression was down-regulated and cell damage was improved.This study indicates that H2S can signifi-cantly improve hypoxia/reoxygenation injury of AEC Ⅱ in rats,and its mechanism may be related to inhibiting the HK2-NLRP3-GSDMD pathway and reducing pyroptosis.
hydrogen sulfiderat alveolar epithelial cell type Ⅱhypoxia/reoxygenation injurythe HK2-NLRP3-GSDMD pathway
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