基于网络药理学与肠道菌群的柏子仁脂肪油改善Aβ_(25-35)诱导小鼠脑损伤的作用研究  

Platycladi Semen oil ameliorates Aβ_(25-35)-induced brain injury in mice based on network pharmacology and gut microbiota

作　　者：曾梦楠[1,2] 曹兵 冯敖梓 郭彭莉 刘萌 张宇涵 李孟 郑晓珂[1,2] ZENG Meng-nan;CAO Bing;FENG Ao-zi;CUO Peng-li;LIU Meng;ZHANG Yu-han;LI Meng;ZHENG Xiao-ke(School of Pharmacy,Henan University of Chinese Medicine,Zhengzhou 450046,China;Engineering and Technology Center for Chinese Medicine Development of Henan Province,Zhengzhou 450046,China;Department of Clinical Research,First Affiliated Hospital of Jinan University,Guangzhou 510632,China)

机构地区：[1]河南中医药大学药学院,河南郑州450046 [2]河南省中药开发工程技术研究中心,河南郑州450046 [3]暨南大学第一附属医院临床研究部,广东广州510632

出　　处：《中国中药杂志》2023年第15期4046-4059,共14页China Journal of Chinese Materia Medica

基　　金：国家重点研发计划项目(2019YFC1708802,2017YFC1702800);国家自然科学基金项目(32200322);中央高校基本科研业务费项目(21622314);河南省中医药科研专项(20-21ZY2152);河南中医药大学博士基金项目(RSBSJJ2019-10)。

摘　　要：研究柏子仁脂肪油(Platycladi Semen oil,SP)对Aβ_(25-35)诱导小鼠脑损伤的保护作用及其机制,为临床治疗阿尔茨海默病(Alzheimer′s disease,AD)提供理论依据。雄性昆明(KM)小鼠随机分为对照组、模型组(脑部注射200μmol·L-1Aβ_(25-35)溶液,0.15μL·g^(-1))、阳性药多奈哌齐组(10 mg·kg^(-1))及柏子仁脂肪油低、高剂量(0.5、1 mL·kg^(-1))组。探究各组小鼠的学习记忆功能、神经元损伤、Aβ1-42/Aβ1-40、p-Tau、细胞凋亡、氧化应激相关指标、免疫细胞及鞘氨醇激酶1(sphingosine kinase 1,SPHK1)/鞘氨醇-1-磷酸(sphingosine-1-phosphate,S1P)/鞘氨醇-1-磷酸受体5(sphingosine-1-phosphate receptor 5,S1PR5)通路相关蛋白或mRNA水平。接着利用气相色谱-质谱联用(gas chromatography-mass spectrometry,GC-MS)分析SP中的成分,并通过网络药理学、肠道菌群(gut mircrobiota,GM)16S rDNA测序及分子对接技术探讨SP干预AD的作用机制。结果显示SP可改善Aβ_(25-35)诱导小鼠的学习记忆功能,减少海马神经元损伤,降低脑内Aβ1-42/Aβ1-40、p-Tau、细胞凋亡和氧化应激相关指标的水平,并维持小鼠机体免疫细胞和GM的稳态。网络药理学和GM测序分析显示,SP对AD的治疗作用与鞘脂通路相关。同时,(Z,Z,Z)-9,12,15-十八碳三烯酸和(Z,Z)-9,12-十八碳二烯酸是SP中含量最高的2种成分,与SPHK1和S1PR5有较好的结合活性。因此,推测SP通过调节肠道菌群,抑制SPHK1/S1P/S1PR5通路来发挥抗凋亡和抗氧化作用,从而改善Aβ_(25-35)诱导小鼠的脑损伤;并且(Z,Z,Z)-9,12,15-十八碳三烯酸和(Z,Z)-9,12-十八碳二烯酸可能是SP发挥抗AD作用的物质基础。The present study aimed to investigate the protective effect and underlying mechanism of Platycladi Semen oil(SP)on Aβ_(25-35)-induced brain injury in mice to provide a theoretical basis for the clinical treatment of Alzheimer′s disease(AD).Male Kunming(KM)mice were randomly divided into a control group,a model group(brain injection of Aβ_(25-35),200μmol·L^(-1),0.15μL·g^(-1)),a positive drug group(donepezil,10 mg·kg^(-1)),and low-and high-dose SP groups(0.5 and 1 mL·kg^(-1)).Learning and memory ability,neuronal damage,levels of Aβ_(1-42)/Aβ_(1-40),p-Tau,related indicators of apoptosis and oxidative stress,and immune cells,and protein and mRNA expression related to the sphingosine kinase 1(SPHK1)/sphingosine-1-phosphate(S1P)/sphingosine-1-phosphate receptor 5(S1PR5)signaling pathway of mice in each group were determined.In addition,compounds in SP were analyzed by gas chromatography-mass spectrometry(GC-MS).The mechanism of SP against AD was investigated by network pharmacology,16S rDNA gene sequencing for gut microbiota(GM),and molecular docking techniques.The results showed that SP could improve the learning and memory function of Aβ_(25-35)-induced mice,reduce hippocampal neuronal damage,decrease the levels of Aβ_(1-42)/Aβ_(1-40),p-Tau,and indicators related to apoptosis and oxidative stress in the brain,and maintain the homeostasis of immune cells and GM.Network pharmacology and sequencing analysis for GM showed that the therapeutic effect of SP on AD was associated with the sphingolipid signaling pathway.Meanwhile,(Z,Z,Z)-9,12,15-octadecatrienoic acid and(Z,Z)-9,12-octadecadienoic acid,the components with the highest content in SP,showed good binding activity to SPHK1 and S1PR5.Therefore,it is inferred that SP exerts anti-apoptosis and antioxidant effects by regulating GM and inhibiting SPHK1/S1P/S1PR5 pathway,thereby improving brain injury induced by Aβ_(25-35)in mice.Moreover,(Z,Z,Z)-9,12,15-octadecatrienoic acid and(Z,Z)-9,12-octadecadienoic acid may be the material basis for the anti-AD ef

关 键 词：阿尔茨海默病 柏子仁脂肪油 肠道菌群 网络药理学 SPHK1/S1P/S1PR5信号通路 

分 类 号：R285.5[医药卫生—中药学]