ACE1通过改善H_(2)O_(2)诱导绒毛膜滋养层细胞铁死亡影响妊娠稽留流产发生的机制研究  

Mechanism of ACE1 affecting the occurrence of missed pregnancy loss by improving H_(2)O_(2)-induced ferroptosis in chorionic trophoblast cells

作　　者：惠淑宁 董利琴 HUI Shuning;DONG Liqin(The First Affiliated Hospital of Xi'an Medical University,Xi'an,Shannxi 710003,China)

机构地区：[1]西安医学院第一附属医院,陕西西安710003

出　　处：《国际检验医学杂志》2025年第8期992-996,共5页International Journal of Laboratory Medicine

摘　　要：目的探讨血管紧张素转换酶1(ACE1)通过改善过氧化氢(H_(2)O_(2))诱导绒毛膜滋养层细胞铁死亡影响妊娠稽留流产发生的机制。方法使用人绒毛膜滋养层细胞HTR-8SVneo进行实验,建立H_(2)O_(2)诱导铁死亡模型,转染ACE1表达载体(pcDNA3.1-ACE1)、ACE1抑制剂(ACE1-siRNA)至细胞,使用实时荧光定量PCR(qPCR)验证转染效果。CCK-8检测各组细胞的细胞活力,DCFH-DA荧光探针法检测活性氧(ROS)水平,Ferrozine法测定细胞中的游离铁离子(Fe^(2+))水平,TBARS法评估各组细胞的脂质过氧化产物丙二醛(MDA)水平,蛋白印迹法(Western blot)检验谷胱甘肽过氧化物酶4(GPX4)、铁蛋白重链1(FTH1)蛋白表达水平。结果CCK-8检测结果显示,H_(2)O_(2)+ACE1处理组的细胞活力较H_(2)O_(2)处理组增强(P<0.05);荧光探针DCFH-DA结果显示,H_(2)O_(2)+ACE1处理组的ROS水平较H_(2)O_(2)处理组降低(P<0.05);Ferrozine法测定细胞中的游离Fe^(2+)水平结果显示,H_(2)O_(2)+ACE1处理组的游离Fe^(2+)水平较H_(2)O_(2)处理组降低(P<0.05);TBARS法评估脂质过氧化产物MDA水平结果显示,H_(2)O_(2)+ACE1处理组的MDA水平较H_(2)O_(2)处理组降低(P<0.05);Western blot结果显示,H_(2)O_(2)+ACE1处理组的GPX4和FTH1蛋白表达水平较H_(2)O_(2)处理组升高(P<0.05)。结论ACE1通过抑制H_(2)O_(2)诱导的铁死亡机制,改善了绒毛膜滋养层细胞的存活率,减少了细胞凋亡和坏死,从而可能在预防和治疗妊娠稽留流产中发挥重要作用。Objective To explore the mechanism of angiotensin-converting enzyme 1(ACE1)affecting the occurrence of missed pregnancy loss by improving H_(2)O_(2)-induced ferroptosis in chorionic trophoblast cells.Methods Human chorionic trophoblast cells HTR-8SVneo were used for experiments,and a H_(2)O_(2)-induced ferroptosis model was established.The cells were transfected with ACE1 expression vector(pcDNA3.1-ACE1)and ACE1 inhibitor(ACE1-siRNA),and real time fluorescence quantitative PCR(qPCR)was used to verify the transfection effect.CCK-8 was used to detect the cell viability of cells in each group,the fluorescent probe DCFH-DA was used to detect reactive oxygen species(ROS)levels,the Ferrozine method was used to measure the level of free Fe^(2+)in cells,the TBARS method was used to evaluate the levels of lipid peroxidation product malondialdehyde(MDA)in cells in each group,and Western blot was used to detect the protein expression levels of glutathione peroxidase 4(GPX4)and ferritin heavy chain 1(FTH1).Results CCK-8 results showed that the cell viability of the H_(2)O_(2)+ACE1 treatment group was significantly higher than that of the H_(2)O_(2)treatment group(P<0.05).The results of the fluorescent probe DCFH-DA showed that the ROS level of the H_(2)O_(2)+ACE1 treatment group was significantly lower than that of the H_(2)O_(2)treatment group(P<0.05).The Ferrozine method measured the free Fe^(2+)level in cells and the results showed that the level of free Fe^(2+)in the H_(2)O_(2)+ACE1 treatment group was decreased when compared with the H_(2)O_(2)treatment group(P<0.05).The TBARS method was used to evaluate the MDA levels of lipid peroxidation products,and the results showed that the MDA levels in the H_(2)O_(2)+ACE1 treatment group were lower than those in the H_(2)O_(2)treatment group(P<0.05);Western blot results showed that the protein expression levels of GPX4 and FTH1 in the H_(2)O_(2)+ACE1 treatment group were significantly increased when compared with the H_(2)O_(2)treatment group(P<0.05).Conclusion ACE1 improves th

关 键 词：血管紧张素转换酶1 过氧化氢 绒毛膜滋养层细胞 铁死亡 妊娠稽留流产 

分 类 号：R714.2[医药卫生—妇产科学]