黄芩素调控SLC7A11/GPX4信号通路改善铁死亡促进骨髓间充质干细胞成骨分化实验研究OA

Objective:To investigate the role of Baicalein(BAI)in inhibiting ferroptosis in bone marrow mesen-chymal stem cells(BMSCs)and promoting osteogenic differentiation through regulating the solute carrier family 7 member 11(SLC7A11)/glutathione peroxidase 4(GPX4)signaling pathway.Methods:A BMSCs ferroptosis and os-teogenic dysfunction model was established by using a high-fat,high-glucose(HGHF)environment combined with the ferroptosis inducer Erastin,followed by intervention with BAI and ferroptosis inhibitor Ferrostatin-1(Fer-1).The experiment was divided into five groups:①Control group(normal culture conditions);② Model group(HGHF environment);③ Ferroptosis inducer group(HGHF+300 μmol/L Erastin);④ Model+BAI group(HGHF+20 μmol/L BAI);⑤ Model+ferroptosis inhibitor group(HGHF+10 μmol/L Fer-1).Cell viability was measured using the CCK-8 assay.Mitochondrial ultrastructure was observed by transmission electron microscopy.Reactive oxy-gen species(ROS),malondialdehyde(MDA),and glutathione(GSH)levels were assessed using DCFH-DA probes and related kits.Mineralized nodule formation was evaluated by alizarin red S staining;and the expression levels of SLC7A11,GPX4,acyl-CoA synthetase long-chain family member 4(ACSL4),ferritin heavy chain 1(FTH1),and os-teogenic markers runt-related transcription factor 2(RUNX2),collagen type Ⅰ alpha 1 chain(COL1A1),and osteo-calcin(OCN)were measured by Western blot.Results:Compared with the control group,the BMSCs cell viability in the model group and ferroptosis inducer group was significantly decreased,with obvious mitochondrial shrinkage,membrane rupture,and cristae reduction(all P＜0.05).Compared with the model group and the ferroptosis inducer group,the cell viability of the model+BAI group was significantly enhanced,the mitochondrial structure was signifi-cantly improved,the intracellular ROS and MDA levels were significantly decreased,and the GSH level was signifi-cantly increased(all P＜0.05).Meanwhile,BAI intervention significantly upregulated the protein expressions of SLC7A11,GPX4,FTH1,RUNX2,COL1A1 and OCN,significantly downregulated the expression of ACSL4,and promoted the formation of mineralized nodules(all P＜0.05).These results indicated that BAI could effectively en-hance the activity and osteogenic differentiation ability of BMSCs,and its effect was similar to that of the model+fer-roptosis inhibitor group.Conclusion:BAI can inhibit ferroptosis in BMSCs and alleviate oxidative stress injury by acti-vating the SLC7A11/GPX4 signaling pathway,thereby restoring their osteogenic differentiation ability.