Effect And Potential Mechanism Of Baicalein On Ferroptosis Induced By Erastin In Osteoblasts

Objective:Previous studies on ferroptosis have been conducted initially,and the traditional Chinese medicine monomer baicalein was screened out as an ideal inhibitor of iron death.In this paper,based on previous studies,we further explored its effect on erastin induced osteoblast iron death as well as the underlying mechanism.Methods:(1)MC3T3-E1 cells,a mouse osteoblast cell line cultured in vitro,were divided into control and experimental groups,which were cultured with different final concentrations of baicalein(0 mmol/L,12.5 mmol/L,25 mmol/L,50 mmol/L,100 mmol/L)for 24 h,and the effects of different concentrations of baicalein on the viability of mouse MC3T3-E1 cells were detected by CCK-8 assay.(2)A murine osteoblast cell line MC3T3-E1 cell model of iron death induced by erastin was established in vitro and divided into control and experimental groups(experimental group:10 mmol/L erastin+0 mmol/L,12.5 mmol/L,25 mmol/L,50 mmol/L,and 100 mmol/L with different final concentrations of baicalein),and the two groups were cultured for 24 h,respectively,and the activity of different concentrations of baicalein on erastin induced iron death mouse MC3T3-E1 cells was examined by CCK-8 assay.(3)The model of iron death of MC3T3-E1 cells induced by erastin was established in vitro.Baicalein(concentration:0 mmol/L、50 mmol/L)was added and cultured for 24 hours,48 hours and 72 hours respectively.The effect of baicalein on the proliferation of MC3T3-E1 cells induced by erastin was detected by CCK-8 assay.(4)The effects of baicalein on the expression of β-catenin and GPX4 are detected by western blot.Results:(1)All these effects of baicalein at concentrations between 12.5 and 100 mmol/L significantly promoted MC3T3-E1 cell proliferation in mouse osteoblast cell line,especially the 50 mmol/L baicalein group showed the most significant effect.(2)In the final concentration of 50 mmol/L baicalein group,significant cell proliferation was observed,that is,baicalein at the concentration of 50 mmol/1 can effectively inhibit ferroptosis.(3)Baicalein at a final concentration of 50 mmol/L inhibited ferroptosis in MC3T3-E1 cells induced by erastin,whereas baicalein at a concentration of 50 mmol/L for 48 h and 72 h did not significantly inhibit ferroptosis.(4)Baicalein(0 mmol/L,50 mmol/L)intervened the ferroptosis of MC3T3-E1 cells induced by erastin for 24 hours.Western blot was used to detected the expression of protein such as GPX4 and β-catenin.The results showed that:compared with the control group,Baicalein group(0 mmol/L)intervened in ferroptosis induced by erastin significantly downed the protein expression of GPX4 and β-catenin.While baicalein group(50 mmol/L)intervened in ferroptosis induced by erastin significantly Increased the protein expression of GPX4 and β-catenin.Conclusion:Baicalein at a certain concentration range is able to promote the proliferation of mouse MC3T3-E1 osteoblasts,and at an appropriate concentration may inhibit iron death by upregulating GPx4 via the Wnt/β-catenin signaling pathway,providing a new direction for the treatment of osteoporosis.