光甘草定衍生物MG01通过抑制铁死亡改善Aβ1-42对HT22细胞的损伤  

Glabridin derivative MG01 protect HT22 cell from Aβ1-42-induced injury by inhibiting ferroptosis

作　　者：孟美辰 王志远 王丽娟 杨寒冰 史佳玮 魏彪 于海洋 周延萌 MENG Meichen;WANG Zhiyuan;WANG Lijuan;YANG Hanbing;SHI Jiawei;WEI Biao;YU Haiyang;ZHOU Yanmeng(Institute of Pharmacology,Shandong First Medical University&Shandong Academy of Medical Sciences,Tai’an 271016,China)

机构地区：[1]山东第一医科大学(山东省医学科学院)药学院(药物研究所),山东泰安271016

出　　处：《中草药》2025年第3期867-876,共10页Chinese Traditional and Herbal Drugs

基　　金：山东省自然科学基金项目(ZR2024MH226);国家自然科学基金项目(82073827)。

摘　　要：目的探究光甘草定衍生物MG01对β-淀粉样多肽1-42(beta-amyloid peptide 1-42,Aβ1-42)诱导的小鼠海马神经元细胞株HT22铁死亡的保护以及预防作用。方法计算机模拟分子对接验证MG01可以与雌激素受体β(etrogen receptor beta,ERβ)以及过氧化物酶体增殖物激活受体γ共激活因子-1α(peroxisome proliferator-activated receptor-γcoactivator-1α,PGC-1α)相互作用;利用Cellcountingkit-8(CCK-8)试剂盒分析MG01对细胞活力的影响并确定最佳给药浓度;活性氧(reactive oxygen species,ROS)试剂盒检测细胞ROS的水平;JC-1线粒体膜电位试剂盒检测线粒体膜电位;细胞免疫荧光染色检测神经元树突标志物微管相关蛋白-2(microtubule-associated protein-2,MAP-2)、铁死亡特异性标志物过氧化物酶3(peroxiredoxin 3,PRDX3)的变化;蛋白质免疫印迹(Western blotting,WB)法检测ERβ、ERα、谷胱甘肽过氧化物酶4(glutathion peroxidase,GPX4)、PRDX3、膜铁转运蛋白1(ferroportin1,FPN1)、铁蛋白(ferritin)、PGC-1α、核因子E2相关因子2(nuclear factor erythroid 2-related factor 2,NRF2)、线粒体转录因子A(transcription factor A,TFAM)蛋白表达水平。结果WB结果表明MG01能显著上调ERβ与PGC-1α的表达(P<0.05、0.01);CCK-8、细胞形态检测与免疫荧光结果表明,与模型组比较,MG01组细胞活力和形态得到明显改善(P<0.05、0.01),且有效浓度显著低于光甘草定(P<0.05)。WB结果表明,MG01能够改善Aβ1-42诱导的细胞铁死亡,改善铁死亡标志蛋白GPX4、PRDX3、FPN1以及ferritin的表达(P<0.01),其作用机制可能与上调ERβ/PGC-1α促进线粒体生物发生,改善线粒体膜电位降低,降低ROS的水平有关。结论MG01介导ERβ/PGC-1α促进线粒体生物发生,改善线粒体功能,抑制铁死亡,发挥神经保护作用。Objective To investigate the protective and preventive effects of glabridin derivative MG01 on ferroptosis induced by beta-amyloid peptide 1-42(Aβ1-42)in HT22 cells.Methods Computer simulation of molecular docking verifies that MG01 can interact with etrogen receptor beta(ERβ)and peroxisome proliferator-activated receptor-γcoactivator-1α(PGC-1α).Cell counting kit-8(CCK-8)was used to analyze the effects of MG01 on cell viability and determine the optimal concentration of MG01.Reactive oxygen species(ROS)detection kit was used to detect the ROS level of cells,and JC-1 mitochondrial membrane potential detection kit was used to detect mitochondrial membrane potential.Cell immunofluorescence(ICC)was used to detect the changes of neuronal dendrite marker microtubule-associated protein-2(MAP-2),ferroptosis specific marker peroxiredoxin 3(PRDX3).Western blotting(WB)was used to detect the expression levels of ERβ,ERα,glutathion peroxidase(GPX 4),PRDX3,ferroportin1(FPN1),ferritin,PGC-1α,nuclear factor erythroid 2-related factor 2(NRF2),and transcription factor A(TFAM).Results WB results showed that MG01 could significantly increase the expression of ERβand PGC-1α(P<0.05,0.01).CCK-8,cell morphology detection and immunofluorescence results showed that compared with the model group,the cell viability and morphology of the MG01 group were significantly improved(P<0.05,0.01),and the effective concentration was significantly lower than that of glabridin(P<0.05).WB results showed that MG01 could improve the cell ferroptosis induced by Aβ1-42,improve the expression of ferroptosis marker proteins GPX4,PRDX3,FPN1,and ferritin(P<0.01),and its mechanism may be related to upregulating ERβ/PGC-1αto promote mitochondrial biogenesis,improving mitochondrial membrane potential,and reducing the level of ROS.Conclusion MG01 mediates ERβ/PGC-1αto promote mitochondrial biogenesis,improve mitochondrial function,inhibit ferroptosis,and play a neuroprotective role.

关 键 词：光甘草定衍生物MG01 阿尔兹海默病 HT22细胞 铁死亡 ERβ/PGC-1α 线粒体 

分 类 号：R285.5[医药卫生—中药学]