文献类型：期刊文献

中文题名：黄芪甲苷对神经毒素损伤PC12细胞的保护作用及分子模拟研究

英文题名：Protective Effect and Molecular Simulation of Astragaloside A on PC12 Cells Damaged by Neurotoxin

作者：刘圆圆[1];彭婷[2];靳晓杰[2];刘永琦[1];姚娟[2]

第一作者：刘圆圆

机构：[1]甘肃中医药大学基础医学院,兰州730000;[2]甘肃中医药大学药学院,兰州730000

第一机构：甘肃中医药大学基础医学院（敦煌医学研究所）

年份：2024

卷号：46

期号：3

起止页码：391

中文期刊名：中国细胞生物学学报

外文期刊名：Chinese Journal of Cell Biology

收录：CSTPCD;;CSCD:【CSCD_E2023_2024】；PubMed;

基金：国家自然科学基金(批准号:82104370);甘肃省自然科学基金(批准号:21JR1RA270);甘肃省高等学校产业支撑计划(批准号:2022CYZC-54)资助的课题。

语种：中文

中文关键词：神经退行性疾病;氧化应激;黄芪甲苷;Keap1-Nrf2;分子对接;分子动力学模拟

外文关键词：neurodegenerative diseases;oxidative stress;Astragaloside A;Keap1-Nrf2;molecular docking;molecular dynamics simulation

摘要：该文研究了黄芪甲苷(Astragaloside A,AS-IV)对神经毒素6-羟多巴胺(6-hydroxydopamine,6-OHDA)损伤PC12细胞的保护作用,并通过分子模拟探究其机制。使用6-OHDA建立PC12细胞损伤模型,通过检测细胞增殖率,培养基上清中乳酸脱氢酶(lactate dehydrogenase,LDH)的释放量,细胞凋亡,细胞内超氧化物歧化酶(superoxide dismutase,SOD)、总谷胱甘肽(glutathione,GSH)的活性,总抗氧化能力(total antioxidant capacity,T-AOC),细胞中核转录因子E2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)蛋白的表达情况,评估AS-IV对PC12细胞的保护作用及机制。将化合物对接至Nrf2的负性调节蛋白Kelch样ECH相关蛋白1(Kelch-like ECH-associated protein-1,Keap1)上,并建立“化合物-靶点”复合物的分子动力学模拟体系,进一步研究其相互作用模式。结果显示,与6-OHDA模型组比较,25、50μmol/L AS-IV可以显著提高6-OHDA损伤后细胞的增殖率,降低LDH的含量,抑制细胞凋亡,并上调总GSH、T-AOC的水平及SOD的活力,提高细胞内总Nrf2的表达水平,上调细胞核Nrf2及下调细胞质Nrf2的表达。将AS-IV对接至Nrf2的负性调节蛋白Keap1上,对接打分为-7.03 kcal/mol,并且复合物体系在50 ns的模拟时间中保持平稳,蛋白质构象稳定。结果表明,AS-IV可以减轻氧化应激所致的PC12细胞损伤,提高细胞的内源性抗氧化能力,其作用机制可能与转录因子Nrf2的激活有关。
This paper proved the protective effect of AS-IV(Astragaloside A)on PC12 cells damaged by the neurotoxin 6-OHDA(6-hydroxydopamine)and explored its mechanism through molecular simulation.The cell damage model of PC12 was established by 6-OHDA.This study examined cell proliferation rate,the re-lease of LDH(lactate dehydrogenase),cell apoptosis,activity of SOD(superoxide dismutase),GSH(glu-tathione),level of T-AOC(total antioxidant capacity)and Nrf2(nuclear factor erythroid 2-related factor 2)protein expression in cells to explore the protective effect of AS-IV on PC12 cells and its mechanism.The compound was docked to Keap1(Kelch-like ECH-associated protein-1),a negative regulatory protein of Nrf2,and the molecu-lar dynamics simulation system of the“compound target”complex was established to further study its interaction mode.The results showed that compared with 6-OHDA model group,25,50μmol/L AS-IV could significantly in-crease the proliferation rate of cells after 6-OHDA injury,reduce the content of LDH,inhibit apoptosis,up-regulate the levels of total GSH,T-AOC and SOD activity,and increase the expression of total Nrf2 in cells.The expression of nuclear Nrf2 was up-regulated and cytoplasm Nrf2 was down-regulated.AS-IV was docked to Keap1,the nega-tive regulatory protein of Nrf2,with a docking score of?7.03 kcal/mol,and the complex system remained stable in the simulation time of 50 ns,and the protein conformation was stable.The results showed that AS-IV could reduce the damage of PC12 cells induced by oxidative stress and improve the endogenous antioxidant capacity of PC12 cells,which might be related to the activation of transcription factor Nrf2.