黑骨藤正丁醇萃取成分治疗大鼠阿尔茨海默病的药效学及作用机制  

n-butanol fraction of ethanol extract of Periploca forrestii Schltr.:its active components,targets and pathways for treating Alcheimer's disease in rats

作　　者：冉念东 刘杰 徐剑 张永萍 郭江涛 RAN Niandong;LIU Jie;XU Jian;ZHANG Yongping;GUO Jiangtao(School of Pharmacy,Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China;National Engineering and Technology Research Center of Miao Medicine,Guiyang 550025,China;Guizhou Engineering Research Center of Chinese Medicine Concoction and Preparation,Guiyang 550025,China)

机构地区：[1]贵州中医药大学药学院,贵州贵阳550025 [2]国家苗药工程技术研究中心,贵州贵阳550025 [3]贵州中药炮制与制剂工程技术研究中心,贵州贵阳550025

出　　处：《南方医科大学学报》2025年第4期785-798,共14页Journal of Southern Medical University

基　　金：国家自然科学基金委员会-贵州省人民政府联合基金(U1812403-2);黔科合基础[2020]1Y372;贵州省高层次创新型人才项目(黔科合平台人才-GCC[2023]037)。

摘　　要：目的探讨黑骨藤正丁醇萃取部位对阿尔茨海默病(AD)的药效学研究及潜在作用机制预测。方法采用超高效液相-四级杆-静电场轨道阱高分辨质谱(UPLC-QE-MS)技术对黑骨藤正丁醇部位的化学成分进行分析鉴定,建立三氯化铝(AlCl3)和D-半乳糖(D-gal)联合诱导50只SPF级雄性AD大鼠模型,使用酶联免疫吸附试验(ELISA)、苏木精-伊红染色(HE)和尼氏染色(Nissl)、免疫组化染色(ICH)、Western blotting等实验为基础,通过网络药理学及分子对接技术预测抗AD潜在作用机制。结果黑骨藤正丁醇萃取部位鉴定出17个化学成分,主要包括苯丙素类、黄酮类、蒽醌类、三萜类、甾体类以及挥发油类。药效学实验结果显示经过黑骨藤正丁醇萃取部位组处理后,大鼠海马中乙酰胆碱酯酶(AChE)含量降低(P<0.05),而乙酰胆碱(ACh)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)的含量增加(P<0.05),在Western blotting实验中神经细胞凋亡因子B淋巴细胞瘤-2(Bcl-2)、磷脂酰肌醇-3激酶(PI3K)、蛋白激酶B(Akt)、磷酸化磷脂酰肌醇-3激酶(p-PI3K)和磷酸化蛋白激酶B(p-Akt)表达上升(P<0.05),而Bax蛋白、半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)表达下降(P<0.01),氧化应激因子核因子红细胞系2相关因子2(Nrf-2)和血红素氧合酶1(HO-1)蛋白表达水平上调(P<0.01),氧化应激反应转录因子(Keap-1)蛋白表达水平下调(P<0.01),脑源性神经营养因子BDNF表达水平均升高(P<0.01),以及β-淀粉样蛋白Aβ、Tau蛋白表达水平均下调(P<0.01),且表现出一定的剂量依赖性。黑骨藤正丁醇萃取部位活性成分与AD共获得TNF、AKT1和ESR1等14个关键靶点。结论初步阐明了黑骨藤正丁醇萃取成分的药效物质基础,并证实了其具有较好的抗AD效果。预测黑骨藤正丁醇萃取部位可通过多组分、多靶点、多途径和多通路发挥抗AD作用。Objective To investigate the active components and possible mechanisms of n-butanol fraction of Periploca forrestii Schltr.ethanol extract for treating Alzheimer's disease(AD).Methods The active components of n-butanol fraction of Periploca forrestii Schltr.ethanol extract were analyzed using UPLC-QE-MS technique.In a SD rat model of AD induced by treatment with AlCl3 and D-gal,the therapeutic effects of low,moderate and high doses of the n-butanol fraction,saline,and donepezil hydrochloride were evaluated using ELISA,HE and Nissl staining,immunohistochemistry and Western blotting.The therapeutic mechanisms of the n-butanol fraction were explored using network pharmacology and molecular docking.Results Seventeen active components were identified from the n-butanol fraction of Periploca forrestii Schltr.ethanol extract,including phenylpropanoids,flavonoids,anthraquinones,triterpenoids,steroids,and volatile oils.In the rat models of AD,treatment with the n-butanol fraction significantly lowed AChE content in the hippocampus,increased the contents of ACh,SOD,CAT,and GSH-Px,enhanced the expressions of neuronal apoptotic factors Bcl-2,PI3K,Akt,p-PI3K,and p-Akt,and reduced the expressions of Bax and caspase-3 proteins.The treatment also dose-dependently up-regulated hippocampal expressions of Nrf-2,HO-1 and BDNF and down-regulated Keap-1,Aβ and Tau expressions.Bioinformatics analysis identified 14 key intersected targets(including TNF,AKT1 and ESR1)between the n-butanol fraction and AD.Conclusion The therapeutic effect of n-butanol fraction of Periploca forrestii Schltr.ethanol extract in AD mice is mediated by its multiple active components that regulate multiple targets and pathways.

关 键 词：UPLC-QE-MS 黑骨藤 正丁醇萃取部位 化学成分 阿尔茨海默病 网络药理学 

分 类 号：R285.5[医药卫生—中药学]