健脾祛痰和血方通过调控铁死亡途径改善氧化低密度脂蛋白诱导的内皮损伤作用机制

Mechanism of Jianpi Qutan Hexue Decoction(健脾祛痰和血方)Improving Endothelial Injury Induced by Oxidative Low-Density Lipoprotein by Regulating Ferroptosis Pathway

目的 探讨健脾祛痰和血方通过调控铁死亡途径改善氧化低密度脂蛋白(oxidized lowdensity lipoprotein, ox-LDL)诱导的内皮损伤的作用机制。方法 制备健脾祛痰和血方冻干粉。内皮细胞分为4组:空白组、模型组、健脾祛痰和血方组(JPQTHXD组)与阿托伐他汀组。CCK-8法检测健脾祛痰和血方对细胞的增殖活力;铁离子染色检测细胞内的铁含量;活性氧(ROS)检测不同组别细胞内活性氧的含量;RT-RCR及Western blot检测铁死亡相关蛋白GPX4、SLC7A11与P53的表达情况。结果 健脾祛痰和血方可显著促进ox-LDL诱导的HUVEC细胞生长,且呈一定的浓度依赖性;健脾祛痰和血方可降低细胞内铁离子与ROS含量;健脾祛痰和血方可以显著升高SLC7A11、GPX4 mRNA与蛋白表达水平,并且能够显著降低P53mRNA与蛋白表达水平。结论 健脾祛痰和血方对ox-LDL诱导的内皮损伤具有一定的保护作用,这种保护作用可能是通过铁死亡途径介导。

Objective To investigate the mechanism of JianpiQutanHexueDecoction(健脾祛痰和血方,JPQTHXD) improving endothelial injury induced by oxidized low density lipoprotein(ox-LDL) by regulating ferroptosis pathway.Methods The lyophilized powder for JPQTHXD was prepared. The endothelial cells were divided into three groups: blank group, model group, JPQTHXD group and atorvastatin group. CCK-8 method was used to detect the proliferation activity of JPQTHXD on cells. Iron ion staining was used to detect iron content in cells. ROS was used to detect the content of reactive oxygen species(ROS) in different groups. RT-RCR and Western blot methods were used to detect the expressions of ferroptosis related proteins GPX4,SLC7A11 and P53.Results JPQTHXDcould significantly promote the growth of ox-LDL-induced human umbilical vein endothelial cells(HUVECs) in a concentration-dependent manner. JPQTHXD can reduce the contents of iron and reactive oxygen species(ROS) in cells. JPQTHXDcan significantly increase the mRNA and protein expression levels of SLC7A11 and GPX4,and significantly reduce the mRNA and protein expression levels of P53.Conclusion JPQTHXDcan protect the endothelial injury induced by ox-LDL,which may be mediated by ferroptosis pathway.