芍药苷对高糖诱导足细胞铁死亡及Nrf2/HMOX-1通路的影响  

Effects of paeoniflorin on high-glucose-induced ferroptosis and Nrf2/HMOX-1 pathway in podocytes

作　　者：余慧娟 项永晶 李正胜 卢晶晶 谢娟 汪丹丹 YU Huijuan;XIANG Yongjing;LI Zhengsheng(The 2th Affiliated Hospital of Guizhou University of TCM,Guizhou,Guiyang 550001,China)

机构地区：[1]贵州中医药大学第二附属医院,贵阳市550001

出　　处：《河北医药》2024年第7期993-996,1001,共5页Hebei Medical Journal

基　　金：贵州省科技计划课题(编号:黔科合基础[2017]1005,贵科合学术新苗[2023]-39号)。

摘　　要：目的探究芍药苷对高糖诱导足细胞铁死亡及Nrf2/HMOX-1通路的影响。方法体外培养小鼠肾足细胞MPC-5,将MPC-5细胞分为对照组、高糖组、高糖+芍药苷低剂量组、高糖+芍药苷高剂量组、高糖+ML385组、高糖+ZNPP组。CCK-8检测细胞增殖;采用流式细胞术检测细胞凋亡率;试剂盒检测Fe^(2+)、超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽(GSH)、活性氧(ROS)水平;RT-PCR法检测核因子E_(2),相关因子2(Nrf2)mRNA、血红素加氧酶(HMOX-1)mRNA;western blot检测Nrf2、HMOX-1、溶质载体家族T成员11(SLC7A11)、谷胱甘肽过氧化物酶4(GPX4)蛋白表达。结果与对照组比较,高糖组MPC-5细胞OD_(450)值(24、48 h)、SOD、GSH水平、SLC7A11、GPX4蛋白水平显著降低,Fe^(2+)、MDA、ROS水平、凋亡率、Nrf2 mRNA、HMOX-1 mRNA表达水平、Nrf2、HMOX-1蛋白水平显著升高(P<0.05);与高糖组相比,高糖+芍药苷低剂量组、高糖+芍药苷高剂量组、高糖+ML385组和高糖+ZNPP组MPC-5细胞OD_(450)值(24、48 h)、SOD、GSH水平、SLC7A11、GPX4蛋白水平显著升高,Fe^(2+)、MDA、ROS水平、凋亡率、Nrf2 mRNA、HMOX-1 mRNA表达、Nrf2、HMOX-1蛋白水平显著降低(P<0.05);与高糖+芍药苷高剂量组比较,高糖+ML385组MPC-5细胞各检测指标差异均无统计学意义(P>0.05),高糖+ZNPP组Nrf2蛋白表达显著升高(P<0.05),其他检查指标差异均无统计学意义(P>0.05)。结论芍药苷通过抑制Nrf2/HMOX-1通路可抑制高糖诱导的足细胞铁死亡。Objective To investigate the effects of paeoniflorin on high-glucose-induced ferroptosis and the nuclear factor E_(2) related factor 2(NRF2)and heme oxygenase-1(HMOX-1)pathway in podocytes.Methods The mouse renal podocyte cell line MPC-5 was cultured in vitro.MPC-5 cells were induced with blank control,high-glucose,high-glucose+low-dose paeoniflorin,high-glucose+high-dose paeoniflorin,high-glucose+ML385 and high-glucose+ZNPP.Cell viability and apoptosis were detected by CCK-8 assay and flow cytometry,respectively.Commercial kits were used to measure Fe^(2+),superoxide dismutase(SOD),malondialdehyde(MDA),glutathione(GSH)and reactive oxygen species(ROS)levels.The mRNA levels of Nrf2(NF-E2-related factor 2)and HMOX-1(heme oxygenase-1)were detected by Quantitative real-time polymerase chain reaction(QRT-PCR).Protein levels of Nrf2,HMOX-1,SLC7A11(olute carrier family 7 member 11)and GPX4(glutathione peroxidase 4)were detected by Western blot.Results Compared with those induced with blank control,MPC-5 cells induced with high-glucose presented significantly lower OD_(450) at 24h and 48h,SOD,GSH and protein levels of SLC7A11 and GPX4,and significantly higher Fe^(2+),MDA,ROS,apoptotic rate,mRNA and protein levels of Nrf2 and HMOX-1(P<0.05).Compared with those induced with high-glucose,MPC-5 cells induced with high-glucose+low-dose paeoniflorin,high-glucose+high-dose paeoniflorin,high-glucose+ML385 and high-glucose+ZNPP presented significantly higher OD_(450) at 24h and 48h,SOD,GSH and protein levels of SLC7A11 and GPX4,and significantly lower Fe^(2+),MDA,ROS,apoptotic rate,mRNA and protein levels of Nrf2 and HMOX-1(P<0.05).There were no significant differences in the above parameters between MPC-5 cells induced with high-glucose+high-dose paeoniflorin and high-glucose+ML385(P>0.05).Except for the significantly higher protein level of Nrf2(P<0.05),there were no significant differences in the remaining parameters between MPC-5 cells induced with high-glucose+high-dose paeoniflorin and high-glucose+ZNPP(P>0.05).Conclusion P

关 键 词：芍药苷 高糖 足细胞 Nrf2/HMOX-1通路 

分 类 号：R587.1[医药卫生—内分泌]