文献类型：期刊文献

中文题名：附子通过AMPK-自噬-铁死亡途径抑制Hela细胞活性

英文题名：Aconiti Lateralis Radix Praeparata Inhibits Hela Cell Viability via the AMPK/Autophagy/Ferroptosis Pathway

作者：杨丽[1];柏虎虎[2];李文霞[1];贺沙沙[1];刘成松[1]

第一作者：杨丽

机构：[1]甘肃中医药大学附属医院药学部,甘肃兰州730030;[2]兰州大学药学院,甘肃兰州730030

第一机构：甘肃中医药大学第二附属医院

年份：2026

卷号：37

期号：2

起止页码：245

中文期刊名：中药新药与临床药理

外文期刊名：Traditional Chinese Drug Research and Clinical Pharmacology

收录：;北大核心:【北大核心2023】；

基金：甘肃省中医药科研课题(GZKP-2022-18);甘肃省科技厅自然科学基金项目(25JRRA240)。

语种：中文

中文关键词：附子;AMPK;自噬;铁死亡;宫颈癌;Hela细胞;细胞活性;小鼠

外文关键词：Aconiti Lateralis Radix Praeparata;AMPK;autophagy;ferroptosis;cervical cancer;Hela cells;cell viability;mice

摘要：目的探讨附子对Hela细胞活性的影响,以及附子导致Hela细胞发生铁死亡的作用机制。方法给小鼠灌胃附子水煎剂,制备附子含药血清;使用附子含药血清替代胎牛血清分别处理人胚胎肾细胞的上皮样细胞HEK 293T细胞和人宫颈癌细胞Hela细胞。使用比色法测定细胞活性;使用酶联免疫吸附法(ELISA)测定细胞中丙二醛(MDA)和谷胱甘肽(GSH)的浓度;使用免疫印迹法测定细胞中谷胱甘肽过氧化物酶4(GPX4)、AMP激活的蛋白激酶(AMPK)和磷酸化的AMP激活蛋白激酶(p-AMPK;特指Thr172位点磷酸化)的蛋白水平;使用AMPK抑制剂BML275和自噬抑制剂3-甲基腺嘌呤(3MA)提前处理Hela细胞,进一步研究AMPK和自噬在附子诱导铁死亡过程中的作用;使用BODIPY 581/591 C11荧光探针测定细胞脂质过氧化水平。结果与对照组比较,附子含药血清不影响HEK 293T细胞活性,却明显降低了Hela细胞的活性(P<0.05,P<0.0001)。附子可使Hela细胞中铁死亡标志物MDA含量升高(P<0.001),GSH含量下降(P<0.001),GPX4蛋白表达明显降低(P<0.001),AMPK及p-AMPK蛋白表达明显升高(P<0.01,P<0.05)。BML275和3MA预处理均能明显增加附子组细胞中GPX4含量(P<0.001,P<0.01)。附子组细胞脂质过氧化程度明显升高(P<0.0001),而BML275和3MA预处理能明显降低附子组细胞脂质过氧化水平(P<0.0001)。结论附子可能通过活化AMPK、诱导细胞自噬促进细胞铁死亡,从而降低Hela细胞活性,在Hela细胞铁死亡过程中发挥重要作用。
Objective To investigate the effect of Aconiti Lateralis Radix Praeparata on the viability of Hela cells and to study the mechanism by which Aconiti Lateralis Radix Praeparata induces ferroptosis in Hela cells.Methods A water decoction of Aconiti Lateralis Radix Praeparata was administered to mice by gavage to prepare drug-containing serum.This Aconiti Lateralis Radix Praeparata-containing serum was used to replace fetal bovine serum for treating human embryonic kidney epithelial cells(HEK 293T)and human cervical cancer cells(Hela).Cell viability was determined by colorimetric assay.Concentrations of malondialdehyde(MDA)and glutathione(GSH)in the cells were measured by enzyme-linked immunosorbent assay(ELISA).Protein levels of glutathione peroxidase 4(GPX4),AMP-activated protein kinase(AMPK),and phosphorylated AMPK(p-AMPK,specifically at Thr172)were determined by Western Blot.Hela cells were pretreated with the AMPK inhibitor BML275 and the autophagy inhibitor 3-methyladenine(3MA)to further explore the roles of AMPK and autophagy in Aconiti Lateralis Radix Praeparata-induced ferroptosis.Lipid peroxidation levels were measured using the BODIPY 581/591 C11 fluorescent probe.Results Compared with the control group,Aconiti Lateralis Radix Praeparata-containing serum did not affect the viability of HEK 293T cells but significantly reduced the viability of Hela cells(P<0.05,P<0.0001).In Hela cells of the Aconiti Lateralis Radix Praeparata group,the level of the ferroptosis marker MDA increased(P<0.001),GSH content decreased(P<0.001),GPX4 level was significantly reduced(P<0.001),and the protein contents of AMPK and p-AMPK were significantly elevated(P<0.01,P<0.05).Pretreatment with either BML-275 or 3MA significantly increased the GPX4 content in the Aconiti Lateralis Radix Praeparata-treated cells(P<0.001,P<0.01).Lipid peroxidation was significantly increased in the Aconiti Lateralis Radix Praeparata group(P<0.0001),and pretreatment with BML-275 or 3-MA significantly reduced this lipid peroxidation level(P<0.0001).Conclusion Aconiti Lateralis Radix Praeparata may reduce Hela cell viability by activating AMPK and inducing autophagy to promote ferroptosis,playing a significant role in the ferroptosis process of Hela cells.