摘要
目的探究红景天苷对阿霉素所致心肌细胞损伤的保护作用及其对铁死亡及核因子E2相关因子2(Nrf2)/血红素加氧酶1(HO-1)信号通路的影响。方法选用人心肌细胞AC16细胞系,分为对照组(正常培养)、模型组(1μmol/L阿霉素处理)、红景天苷组(1μmol/L阿霉素+100μmol/L红景天苷处理)和抑制剂组(1μmol/L阿霉素+100μmol/L红景天苷+10μmol/L Nrf2抑制剂ML385处理)。采用CCK-8法检测细胞活力,使用铁离子检测试剂盒检测细胞内铁离子含量,使用丙二醛(MDA)检测试剂盒测定细胞内MDA含量,通过流式细胞仪检测细胞内活性氧(ROS)水平;Western blot和qPCR分别测定细胞Nrf2、HO-1、谷胱甘肽过氧化酶4(GPX4)、溶质载体家族7成员11(SLC7A11)和Kelch样ECH相关蛋白1(Keap1)的蛋白及mRNA表达水平。结果(1)4组细胞的细胞活力、铁离子含量、MDA含量及ROS荧光强度均差异有统计学意义(F=24.788、18.940、39.366、5.767,均P<0.05)。红景天苷组细胞活力[(80.35±10.68)%]高于模型组[(50.32±12.32)%]和抑制剂组[(61.26±8.65)%](均P<0.05);红景天苷组铁离子含量、MDA含量及ROS荧光强度均低于模型组和抑制剂组(均P<0.05)。(2)qPCR结果显示,4组细胞HO-1、GPX4、SLC7A11和Keap1的mRNA水平均差异有统计学意义(F=48.107、23.978、53.938、127.176,均P<0.001);红景天苷组细胞HO-1、GPX4、SLC7A11的mRNA水平均高于模型组和抑制剂组(均P<0.05),Keap1的m RNA水平均低于模型组和抑制剂组(均P<0.05)。(3)Western blot结果显示,4组细胞Nrf2、HO-1、GPX4、SLC7A11和Keap1蛋白表达均差异有统计学意义(F=730.392、596.727、147.094、106.472、131.976,均P<0.05)。红景天苷组Nrf2、HO-1、GPX4和SLC7A11蛋白表达水平均高于模型组(均P<0.05),Keap1蛋白表达水平低于模型组(P<0.05);抑制剂组Nrf2、HO-1、SLC7A11蛋白表达水平低于红景天苷组(均P<0.05),Keap1蛋白表达水平高于红景天苷组(P<0.05)。结论红景天苷能有效减轻阿霉素诱导的心肌细胞损伤,改善细胞活力,改善铁死亡相关指标。
Objective To explore the protective effect of salidroside on doxorubicin-induced myocardial cell injury and its impact on ferroptosis and the nuclear factor-erythroid 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathway.Methods The human cardiomyocyte AC16 cell lines were selected and divided into the control group(normal culture),the model group(1μmol/L doxorubicin treatment),the salidroside group(1μmol/L doxorubicin plus 100μmol/L salidroside treatment),and the inhibitor group(1μmol/L doxorubicin plus 100μmol/L salidroside plus 10μmol/L Nrf2 inhibitor ML385 treatment).Cell viability was assessed using the CCK-8 assay.Intracellular iron ion content was measured using an iron ion detection kit.The level of malondialdehyde(MDA)in cells was determined using a MDA detection kit.Reactive oxygen species(ROS)levels in cells were detected using flow cytometry.Western blot and qPCR were used to measure the protein and mRNA expression levels of Nrf2,HO-1,glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(SLC7A11),and Kelchlike ECH-associated protein 1(Keap1)in cells.Results(1)The cell viability,iron ion content,MDA content,and ROS fluores cence intensity in the four groups showed statistically significant differences(F=24.788,18.940,39.366,5.767,all P<0.05).The cell viability in the salidroside group((80.35±10.68)%)was higher than that in the model group((50.32±12.32)%)and the inhibitor group((61.26±8.65)%)(both P<0.05);the iron ion content,MDA content and ROS fluorescence intensity in the salidroside group were lower than those in the model group and the inhibitor group(all P<0.05).(2)PCR results showed that the mRNA levels of HO-1,GPX4,SLC7A11 and Keap1 in the four groups were all statistically significant different(F=48.107,23.978,53.938,127.176,all P<0.001).The mRNA levels of HO-1,GPX4 and SLC7A11 in the salidroside group were higher than those in the model group and the inhibitor groups(all P<0.05),while the mRNA level of Keap1 in the salidroside group was lower than that in the model group and the inhibitor group(both P<0.05).(3)Western blot results showed that the protein expression levels of Nrf2,HO-1,GPX4,SLC7A11 and Keap1 in the four groups were all statistically significant different(F=730.392,596.727,147.094,106.472,131.976,all P<0.05).The protein expression levels of Nrf2,HO-1,GPX4 and SLC7A11 in the salidroside group were higher than those in the model group(all P<0.05),and the protein expression level of Keap1 was lower than that in the model group(P<0.05).The protein expression levels of Nrf2,HO-1 and SLC7A11 in the inhibitor group were lower than those in the salidroside group(all P<0.05),while the protein expression level of Keap1 was higher than that in the salidroside group(P<0.05).Conclusion Salidroside can effectively reduce doxorubicin-induced myocardial cell injury,improve cell viability,and inhibit ferroptosis-related indicators.
作者
张永杰
王星
张明磊
齐贵彬
张志良
高建步
ZHANG Yongjie;WANG Xing;ZHANG Minglei;QI Guibin;ZHANG Zhiliang;GAO Jianbu(Department of Cardiology,Nanyang Central Hospital,Nanyang 473000,China)
出处
《中国药物应用与监测》
2025年第9期1599-1603,共5页
Chinese Journal of Drug Application and Monitoring
基金
河南省医学科技攻关联合共建项目(LHGJ20221049)
南阳市科技发展计划项目(23KJGG126)。
