栀子苷通过激活SIRT3信号通路抑制铁死亡减轻内皮细胞炎症损伤的研究OA
Inhibition of ferroptosis by geniposide via activation of the SIRT3 signaling pathway to alleviate inflammatory injury in endothelial cells
目的 观察栀子苷(geniposide,GE)对脂多糖(lipopolysaccharide,LPS)诱导的人冠状动脉内皮细胞(human coronary artery endothelial cell,HCAEC)炎症损伤的作用及机制.方法 将HCAEC随机分为4组:对照组不予处理,模型组给予LPS(20 ng/mL)处理24 h,GE组给予GE(30 μg/mL)预处理12 h后使用LPS(20 ng/mL)处理24 h,沉默信息调节因子3(silent information regulaton 3,SIRT3)抑制剂组给予GE(40 μg/mL)与SIRT3特异性抑制剂3-TYP共同预处理12 h后使用LPS(20 ng/mL)处理24 h.采用CCK-8法检测细胞活力,试剂盒检测细胞培养基中乳酸脱氢酶(lactate dehydrogenase,LDH)活性及细胞中丙二醛(malondialdehyde,MDA)、谷胱甘肽(glutathione,GSH)、亚铁离子(Fe2+)水平,DCFH-DA荧光探针法检测活性氧(reactive oxygen species,ROS)水平,Western blot法检测SIRT3、谷胱甘肽过氧化物酶4(glutathuone peroxidase 4,GPX4)、溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11)、转铁蛋白受体1(transferrin receptor 1,TfR1)蛋白表达水平,免疫荧光染色检测SIRT3表达.结果 与对照组相比,模型组细胞活力、GSH含量、SIRT3荧光强度及GPX4、SLC7A11、SIRT3蛋白表达降低(P<0.05),LDH活性、TfR1蛋白表达及MDA、Fe2+、ROS含量升高(P<0.05);与模型组相比,GE组细胞活力、GSH含量、SIRT3荧光强度及GPX4、SLC7A11、SIRT3蛋白表达升高(P<0.05),LDH活性、TfR1蛋白表达及MDA、Fe2+、ROS含量降低(P<0.05);与GE组比较,SIRT3抑制剂组细胞活力、GSH含量、SIRT3荧光强度及GPX4、SLC7A11、SIRT3蛋白表达降低(P<0.05),LDH活性、TfR1蛋白表达及MDA、Fe2+、ROS含量升高(P<0.05).结论 GE可改善LPS诱导的HCAEC炎症损伤,其机制可能与抑制细胞铁死亡及激活SIRT3信号通路有关.
Objective To study the effect and mechanism of geniposide(GE)on lipopolysaccharide(LPS)-induced inflammatory injury in human coronary artery endothelial cells(HCAECs).Methods HCAECs were randomly assigned to four groups:control(no treatment);model(LPS 20 ng/mL for 24 hours);GE(GE 30 μg/mL pretreatment for 12 hours,followed by LPS 20 ng/mL for 24 hours);and SIRT3 inhibitor(GE 40 μg/mL plus the SIRT3-specific inhibitor 3-TYP pretreatment for 12 hours,followed by LPS 20 ng/mL for 24 hours).Cell viability was assessed by CCK-8 assay.Lactate dehydrogenase(LDH)activity in culture medium and intracellular malondialdehyde(MDA),glutathione(GSH),and ferrous ion(Fe2+)levels were measured using commercial kits.Reactive oxygen species(ROS)were evaluated by DCFH-DA fluorescent probe method.Western blotting detected the protein expression of silent information regulator 3(SIRT3),glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(SLC7A11),and transferrin receptor 1(TfR1).SIRT3 expression was examined by immunofluorescence staining.Results Compared with the control group,the model group showed decreased cell viability,GSH content,SIRT3 immunofluorescence intensity,and GPX4,SLC7A11,and SIRT3 protein expression(P<0.05),with increased LDH activity,TfR1 protein expression,and MDA,Fe2+,and ROS levels(P<0.05).Compared with the model group,the GE group exhibited increased cell viability,GSH content,SIRT3 immunofluorescence intensity,and GPX4,SLC7A11,and SIRT3 protein expression(P<0.05),and decreased LDH activity,TfR1 protein expression,and MDA,Fe2+,and ROS levels(P<0.05).Compared with the GE group,the SIRT3 inhibitor group showed reduced cell viability,GSH content,SIRT3 immunofluorescence intensity,and GPX4,SLC7A11,and SIRT3 protein expression(P<0.05),along with elevated LDH activity,TfR1 protein expression,and MDA,Fe2+,and ROS levels(P<0.05).Conclusions Geniposide alleviates LPS-induced inflammatory injury in HCAECs,and the mechanism may involve inhibition of ferroptosis and activation of SIRT3 signaling.
孙文婷;赵苗苗;石曌玲
陕西中医药大学,陕西 咸阳 712000陕西中医药大学,陕西 咸阳 712000陕西中医药大学第二附属医院,陕西 咸阳 712000
栀子苷沉默信息调节因子3铁死亡脂多糖人冠状动脉内皮细胞
GeniposideSilent information regulator 3FerroptosisLipopolysaccharideHuman coronary artery endothelial cell
《中国当代儿科杂志》 2026 (4)
486-492,7
国家自然科学基金面上项目(82474575)国家自然科学基金青年科学基金项目(82205188).
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