基于血浆代谢组学研究三七皂苷R1改善神经炎症的分子作用机制  

Molecular Mechanism of Anti-neuroinflammation effect of Notoginsenside R1 Based on Plasma Metabolomics

作　　者：张燕燕 林楠 吴迪 李叶 魏艺聪[1] ZHANG Yan-yan;LIN Nan;WU Di;LI Ye;WEI Yi-cong(College of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China)

机构地区：[1]福建中医药大学药学院,福建福州350122

出　　处：《中国现代中药》2024年第7期1186-1196,共11页Modern Chinese Medicine

基　　金：福建省自然科学基金项目(2018J01869)。

摘　　要：目的:通过血浆代谢组学研究三七皂苷R1(notoginsenside R1,NGR1)治疗神经炎症的分子作用机制。方法:采用腹腔注射0.83 mg‧kg^(-1)脂多糖(LPS)构建小鼠神经炎症模型,应用NGR1(30 mg‧kg^(-1))进行给药治疗,眼眶静脉丛取血并分离血浆。在使用苏木素-伊红(HE)染色法检测小鼠脑组织损伤情况、采用实时荧光定量聚合酶链式反应(qRT-PCR)技术检测神经炎症相关指标及酶联免疫吸附反应(ELISA)技术分析血清炎症因子变化的基础上,应用液相色谱-质谱法(LC-MS)对各组小鼠血浆样品进行代谢组学分析,通过人类代谢组数据库(HMDB)、京都基因与基因组百科全书(KEGG)数据库进行差异内源代谢物的鉴定,并应用MetaboAnalyst和Metscape数据库进一步分析差异内源代谢物的关键代谢途径,最后通过qRT-PCR检测调控关键代谢途径基因表达水平的变化。结果:NGR1能够明显改善小鼠脑组织神经损伤,明显降低中枢神经炎症因子离子化钙结合适配分子1(Iba-1)、白细胞介素-1β(IL-1β)、IL-6的mRNA水平,显著提高转化生长因子-β(TGF-β)的mRNA水平,显著降低血清炎症因子IL-6、肿瘤坏死因子-α(TNF-α)的表达水平。此外,代谢组学发现模型组与给药组之间小鼠血浆中皮质酮、吲哚丙酮酸、泛酸、12S-羟基5Z,8Z,10E,14Z-二十碳四烯酸、PC(18:3(9Z,12Z,15Z)/16:1(9Z))等22个差异内源代谢物发生明显变化,主要涉及甘油磷脂代谢、亚油酸代谢及类固醇激素生物合成等代谢通路。qRT-PCR结果显示,给药后能显著逆转关键代谢途径中2A型磷脂酶A2(PLA2G2A)、1B型磷脂酶A2(PLA2G1B)、12A型磷脂酶A2(PLA2G12A)、醛酮还原酶1D1(AKR1D1)、羟基类固醇11β脱氢酶1(HSD11B1)、羟基类固醇11β脱氢酶2(HSD11B2)mRNA表达水平。结论:NGR1对LPS诱导的神经炎症具有治疗作用,并可能通过调控皮质酮等关键代谢物及甘油磷脂代谢、亚油酸代谢及类固醇激素生物合成等关键途径发挥�Objective:This study applied plasma metabolomics to analyze the mechanism of notoginsenside R1(NGR1)against neuroinflammation.Methods:The neuroinflammation model was constructed by intraperitoneal injection of lipopolysaccharide(LPS,0.83 mg‧kg^(-1)),and NGR1(30 mg‧kg^(-1))was used for treatment.Blood was collected from the orbital venous plexus of mice,and plasma was isolated.Hematoxylin-Eosin(HE)staining was employed for detecting the damage to the brain tissue of mice,and real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was employed to detect neuroinflammation-related indicators.Enzyme-linked immunosorbent reaction(ELISA)was used to analyze the changes in inflammatory factors.Metabolomics analysis of mouse plasma samples was performed by liquid chromatography-mass spectrometry(LC-MS),and differential endogenous metabolites were identified by Human Metabolome Database(HMDB)and Kyoto Encyclopedia of Genes and Genomes(KEGG).MetaboAnalyst and Metscape databases were used to further analyze the key metabolic pathways of differential endogenous metabolites.Finally,qRT-PCR was used to detect the changes in gene expression levels regulating key metabolic pathways.Results:The experiments showed that NGR1 could significantly improve nerve injury in the brain tissue of mice and significantly reduce the mRNA levels of central nervous inflammatory factor calcium-ionized binding adaptation molecule 1(Iba^(-1)),interleukin^(-1)β(IL^(-1)β),and IL-6.It could significantly increase the mRNA level of transforming growth factor-β(TGF-β)and decrease the expression levels of serum inflammatory factors IL-6 and TNF-α.Furthermore,metabolomics results revealed that 22 different endogenous metabolites such as corticosterone,indole-3-pyruvic acid,D-(+)-pantothenic acid,12S-hydroxy-5Z,8Z,10E,14Z-eicosatetraenoic acid,and PC(18:3(9Z,12Z,15Z)/16:1(9Z))in plasma of mice were significantly changed between the model group and the administration group.They were mainly involved in glycerophospholipid metabolism,lino

关 键 词：血浆代谢组学 分子作用机制 三七皂苷R1 神经炎症 

分 类 号：R285.5[医药卫生—中药学]