文献类型：期刊文献

中文题名：甘草查尔酮A对神经毒素损伤PC12细胞的保护作用及分子动力学模拟研究

英文题名：Protective effect and molecular dynamics simulation of licochalcone A on PC12 cells damaged by neurotoxin

作者：姚娟[1];刘圆圆[2];张敏[1];刘雪枫[1];靳晓杰[1]

第一作者：姚娟

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

第一机构：甘肃中医药大学药学院（西北中藏药协同创新中心办公室）

年份：2024

卷号：36

期号：12

起止页码：2008

中文期刊名：天然产物研究与开发

外文期刊名：Natural Product Research and Development

收录：CSTPCD;;北大核心:【北大核心2023】；CSCD:【CSCD2023_2024】；

基金：甘肃省自然科学基金(21JR1RA270);2022年陇原青年创新创业人才(个人)项目。

语种：中文

中文关键词：甘草查尔酮A;氧化损伤;核因子E2相关因子;Keap1;神经保护;ADMET

外文关键词：licochalcone A;oxidative damage;nuclear factor erythroid-2 related factor 2;Keap1;neuroprotection;ADMET

摘要：研究甘草查尔酮A对神经毒素6-羟基多巴胺(6-hydroxydopamine,6-OHDA)损伤PC12细胞的保护作用并采用分子模拟探究其机制。首先测定甘草查尔酮A对体外ABTS及DPPH自由基的清除活性。其次采用MTT法、测定细胞培养基中乳酸脱氢酶(lactate dehydrogenase,LDH)的含量评价甘草查尔酮A对6-OHDA损伤PC12细胞的保护作用;2′,7′-二氯二氢荧光素二乙酸酯(2′,7′-dichlorodihydrofluorescein diacetate,DCFH-DA)染色测定甘草查尔酮A对6-OHDA损伤PC12细胞活性氧的影响;通过检测各组细胞中总谷胱甘肽(glutataione,GSH)、超氧化物歧化酶(superoxide dismutase,SOD)的含量及总抗氧化能力(total antioxidant capacity,T-AOC)评价内源性的抗氧化能力;Western blot分析甘草查尔酮A对核因子E2相关因子(nuclear factor erythroid-2 related factor 2,Nrf2)蛋白表达的影响。最后采用分子对接与分子模拟方法从分子层面研究甘草查尔酮A作用于Keap1蛋白的结合位点及结合能力,并预测甘草查尔酮A的吸收、分布、代谢、排泄和毒性(absorption,distribution,metabolism,excretion,toxicity,ADMET)参数。结果表明甘草查尔酮A具有显著的体外自由基清除活性;甘草查尔酮A处理PC12细胞能显著提高6-OHDA损伤环境下PC12细胞的活性、清除细胞中的部分活性氧、提高6-OHDA损伤环境下PC12细胞中总GSH、SOD的含量及T-AOC;Western blot结果表明甘草查尔酮A可提高Nrf2蛋白的表达;分子对接及分子动力学模拟结果显示甘草查尔酮A与Keap1可通过共价键稳定结合,ADMET预测提示甘草查尔酮A具有一定的成药潜力。甘草查尔酮A对6-OHDA损伤的PC12细胞具有一定的保护作用,其机制可能与参与上调生物系统内源性的抗氧化作用相关。
This study investigated the protective effect of licochalcone A(LCA)on PC12 cells damaged by the neurotoxin 6-hydroxy-dopamine(6-OHDA),elucidated its mechanism through molecular simulation.At first,the study determined the scavenging activity of LCA on ABTS and DPPH free radicals in vitro.Secondly,the experiment measured the content of lactate dehydrogenase(LDH)in cell medium and employed the MTT assay to evaluate the protective efficacy of LCA against 6-OHDA-induced damage in PC12 cells.2′,7′-Dichlorodihydrofluorescein diacetate(DCFH-DA)staining was used to determine the effect of LCA on reactive oxygen species in PC12 cells injured by 6-OHDA.The contents of total glutathione(GSH),superoxide dismutase(SOD)and total antioxidant capacity(T-AOC)in each group were detected to evaluate the endogenous antioxidant capacity.The effect of LCA on the expression of nuclear factor erythroid-2 related factor 2(Nrf2)protein was analyzed by Western blot.Finally,molecular docking and simulation methods were utilized to study the binding site and ability of LCA on Keap1 protein,and calculate and analysis its absorption,distribution,metabolism,excretion,toxicity(ADMET).The results indicated that LCA possesses significant free radical scavenging activity in vitro.Treatment with LCA markedly enhanced PC12 cell viability under 6-OHDA stress,diminished reactive oxygen species production,and elevated GSH,SOD,and T-AOC levels under these conditions.Furthermore,Western blot analysis revealed that LCA treatment increased the expression of Nrf2 protein.Molecular docking and dynamic simulations confirmed stable covalent bonding between LCA and Keap1 proteins,ADMET prediction suggesting that LCA could be a promising therapeutic agent.These observations suggest that LCA confers protection to PC12 cells damaged by 6-OHDA by modulating endogenous antioxidant responses within biological systems.