铁死亡在炎症微环境中抑制牙周膜干细胞增殖及成骨分化的作用

Role Ferroptosis in the Inhibition of Proliferation and Osteogenic Differentiation of Periodontal Membrane Stem Cell under Inflammatory Microenvironment

目的:研究铁死亡在炎症微环境中抑制牙周膜干细胞(Periodontal ligament stem cells,PDLSCs)增殖及成骨分化的作用。方法:培养PDLSCs,将不含药物培养基处理的细胞作为对照组,炎症组用含有10μg/L TNF-α的培养基处理,铁死亡激动剂组用含有10μg/L TNF-α及10μmol/L Erastin的培养基处理,铁死亡抑制剂组用含有10μg/L TNF-α及1μmol/L Ferrostatin-1的培养基处理。检测各组细胞增殖活力、转铁蛋白受体1 (Transferr in receptor 1,TfR1)、谷胱甘肽过氧化物酶(Glutathione peroxidase,GPx4)的表达水平及成骨诱导分化后的钙结节数量、碱性磷酸酶(Alkaline phosphatase,ALP)活力、成骨标志基因Runt相关转录因子2 (Runt-related transcription factor 2,Runx2)、骨钙素(Osteocalcin,OCN)、骨桥蛋白(Osteopontin,OPN)的表达水平。结果:炎症组TfR1表达水平高于对照组、GPx4表达水平低于对照组(P<0.05)。炎症组处理第3、5、7d时的增殖活力,成骨诱导第21 d时的钙结节OD540水平,成骨诱导第7d时的Runx2、OCN、OPN表达水平均低于对照组(P<0.05);铁死亡激动剂组的上述指标低于炎症组、铁死亡抑制剂组的上述指标高于炎症组(P<0.05)。结论:炎症微环境下铁死亡的激活抑制PDLSCs的增殖及成骨分化。

Objective To study the role and mechanism of ferroptosis in the inhibition of proliferation and osteogenic differentiation of periodontal membrane stem cells(PDLSCs)under inflammatory microenvironment.Methods PDLSCs were cultured,cells treated with drug free medium served as control group,inflammation group was treated with medium containing 10μg/L TNF-α,ferroptosis agonist group was treated with medium containing 10μg/L TNF-αand 10μmol/L erastin,and ferroptosis inhibitor group was treated with medium containing 10μg/L TNF-αand 1μmol/L ferrostatin-1.The expression levels of cell proliferation viability,transferrin receptor 1(TfR1),glutathione peroxidase(GPx4),the number of calcium nodules,the alkaline phosphatase(ALP)viability,the expression levels of Runt-related transcription factor 2(Runx 2),osteocalcin(OCN),and osteopontin(OPN)after osteogenic induction were measured.Results TfR 1 expression level of the inflammation group was higher than the control group,GPx4 expression level was lower the control group(P<0.05).The proliferative activity of the inflammatory group at the 3rd,5th and 7th day,the level of calcium nodules OD540 at the 21st day of osteogenesis induction,and the expression levels of Runx2,OCN and OPN at the 7th day of osteogenesis induction were lower than those of the control group(P<0.05).The above indexes in the iron death agonist group were lower than those in the inflammation group,and those in the iron death inhibitor group were higher than those in the inflammation group(P<0.05).Conclusion The activation of ferroptosis in the inflammatory microenvironment suppresses PDLSCs proliferation and osteogenic differentiation.