中药药理与临床2025，Vol.41Issue(2)：47-53,7.
丹参葛根提取物抑制Erastin诱导的HT22细胞铁死亡
Danshen(丹参)-Gegen(葛根)Extract Inhibits Erastin-Induced Ferroptosis of HT22 Cells
摘要
Abstract
Objective:To investigate the protective effect of Danshen(丹参)-Gegen(葛根)extract on erastin-induced ferroptosis in mouse hipp-ocampal neuron(HT22)cells.Methods:The water extraction method was employed to prepare the Danshen-Gegen extract.The HT22 cell model of erastin-induced ferroptosis was established in vitro.HT22 cells in the logarithmic growth phase were classified into control,model(erastin,5 μmol/L),Danshen-Gegen extract(10,30,and 100 μg/mL),and ferroptosis inhibitor Fer-1(2 μmol/L)groups.Cell viability was measured by the methyl thiazolyl tetrazolium(MTT)assay.The release of lactate dehydrogenase(LDH)in the cell supernatant and the levels of intracellular iron ion(Fe2+),malondialdehyde(MDA),and glutathione(GSH)were determined by spectrophotometry.The ultra-structure of mitochondria was observed by transmission electron microscopy.The levels of interleukin(IL)-6,IL-1β,and tumor necrosis factor(TNF)-α associated with ferroptosis were determined by enzyme-linked immunosorbent assay.Lipid peroxidation and reactive oxygen species(ROS)levels were detected by the BODIPYTM581/591 C11 assay.The immunofluorescence assay was used to detect the expression of gluta-thione peroxidase 4(GPX4),nuclear factor erythroid 2-related factor-2(NRF2),and acyl-coenzyme A synthase long-chain family member 4(ACSL4).Western blotting was employed to determine the expression levels of ferroptosis-related proteins,i.e.,solute carrier family 7 mem-ber 11(SLC7A11,also known as xCT)and transferrin receptor 1(TFR1).Results:Compared with the control group,the model group showed decreased cell survival rate(P<0.01),increased LDH release and intracellular Fe2+level(P<0.01),and altered mitochondrial mor-phology.Compared with the model group,Danshen-Gegen extract groups showed increased cell survival rate(P<0.05,P<0.01),reduced LDH release and MDA,intracellular Fe2+,and ROS levels(P<0.05,P<0.01),and elevated GSH level(P<0.05,P<0.01).In addition,Danshen-Gegen extract groups showed up-regulated protein levels of NRF2,GPX4,and xCT and down-regulated protein levels of ACSL4 and TFR1(P<0.05,P<0.01).Conclusion:Danshen-Gegen extract may exert the neuroprotective effect by inhibiting erastin-induced ferroptosis in HT22 cells.关键词
丹参葛根提取物/缺血性卒中/铁死亡/谷胱甘肽过氧化酶 4/核因子红细胞-2相关因子-2NRF2/转铁蛋白受体 1Key words
Danshen(丹参)-Gegen(葛根)extract/Ischemic stroke/Ferroptosis/Glutathione peroxidase 4(GPX4)/Nuclear factor ery-throid 2-related factor-2(NRF2)/Transferrin receptor 1(TFR1)引用本文复制引用
王艳,刘茵,贺晓丽..丹参葛根提取物抑制Erastin诱导的HT22细胞铁死亡[J].中药药理与临床,2025,41(2):47-53,7.基金项目
国家自然科学基金资助项目(编号:82274407). （编号:82274407）
