基于UPLC-Q/TOF-MS的黄精不同炮制品干预AD大鼠尿液代谢组学研究  

Different processed products of Polygonati Rhizoma treat Alzheimer′s disease in rats:urine metabolomics based on UPLC-Q/TOF-MS

作　　者：朱娜 朱徐东 杨毅生 严斐霞 张鹏[1,2] 万林春 吴毅 黄丽萍 ZHU Na;ZHU Xu-dong;YANG Yi-sheng;YAN Fei-xia;ZHANG Peng;WAN Lin-chun;WU Yi;HUANG Li-ping(Jiangxi University of Chinese Medicine,Nanchang 330004,China;NMPA Key Laboratory of Quality Evaluation of Traditional Chinese Patent Medicine,Jiangxi Provincial Engineering Research Center for Drug and Medical Device Quality,Jiangxi Institute for Drug Control,Nanchang 330029,China)

机构地区：[1]江西中医药大学,江西南昌330004 [2]江西省药品检验检测研究院,国家药品监督管理局中成药质量评价重点实验室,江西省药品与医疗器械质量工程技术研究中心,江西南昌330029

出　　处：《中国中药杂志》2023年第24期6663-6675,共13页China Journal of Chinese Materia Medica

基　　金：江西省重点研发计划项目(20192BBG70071,20202BBGL73006);江西省主要学科学术和技术带头人项目(20182BCB22005);2023年全国中药特色技术传承人才培训项目(T20234832005);江西省中药饮片炮制规范及我省道地药材饮片品质提升研究项目(20223BBG71001)。

摘　　要：探讨黄精不同炮制品(黑豆黄精、炆黄精)干预阿尔茨海默病(Alzheimer′s disease,AD)模型大鼠尿液代谢产物的影响。将60只SPF级雄性SD大鼠随机分为假手术组、模型组、多奈哌齐组、黑豆黄精组、炆黄精组,每组12只,各组进行腹腔注射D-半乳糖(100 mg·kg^(-1)),假手术组腹腔注射等容积生理盐水。1次/d,持续给药7周。D-半乳糖注射3周后,进行双侧海马Aβ25-35注射进行造模。第2周开始灌胃给药,模型组与假手术组同体积的双蒸水,其余组给予对应的药物(10 mL·kg^(-1)),1次/d,连续灌胃35 d。观察各组大鼠记忆行为学以及海马组织病理学。利用超高效液相色谱-串联三重四极杆飞行时间质谱(UPLC-Q/TOF-MS)技术对尿液进行非靶向代谢物检测,并结合主成分分析(PCA)、正交偏最小二乘法判别分析(OPLS-DA)处理数据以筛选差异代谢物、潜在生物标志物,并进行代谢通路富集分析。结果表明,AD大鼠经过黑豆黄精、炆黄精治疗后新物体识别指数升高,Morris水迷宫实验中逃避潜伏期缩短、穿越平台次数增加。苏木精-伊红(hematoxylin-eosin,HE)染色结果,给药组海马组织细胞排列紧密,海马组织中白细胞介素-6(interleukin-6,IL-6)含量显著降低(P<0.01),肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)相对降低,其中黑豆黄精组更为明显,具显著性差异(P<0.05)。经过筛选处理,得到潜在生物标志物15个,代谢物主要涉及2条代谢通路:双香豆素作用途径、吡罗昔康作用途径。因此,黑豆黄精、炆黄精对AD具有干预缓解作用,其作用机制可能与双香豆素代谢、吡罗昔康代谢有关。The study investigated the effects of different processed products of Polygonati Rhizoma(black bean-processed Polygonati Rhizoma,BBPR;stewed Polygonati Rhizoma,SPR)on the urinary metabolites in a rat model of Alzheimer′s disease(AD).Sixty SPF-grade male SD rats were randomized into a control group,a model group,a donepezil group,a BBPR group,and a SPR group,with twelve rats in each group.Other groups except the control group were administrated with D-galactose injection(100 mg·kg^(-1))once a day for seven weeks.The control group was administrated with an equal volume of normal saline once a day for seven consecutive weeks.After three weeks of D-galactose injection,bilateral hippocampal Aβ25-35 injections were performed for modeling.The rats were administrated with corresponding drugs(10 mL·kg^(-1))by gavage since week 2,and the rats in the model and control group with an equal volume of double distilled water once a day for 35 continuous days.The memory behaviour and pathological changes in the hippocampal tissue were observed.The untargeted metabolites in the urine were detected by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q/TOF-MS).Principal component analysis(PCA)and orthogonal partial least square-discriminant analysis(OPLS-DA)were employed to characterize and screen differential metabolites and potential biomarkers,for which the metabolic pathway enrichment analysis was conducted.The results indicated that BBPR and SPR increased the new object recognition index,shortened the escape latency,and increased the times of crossing the platform of AD rats in the Morris water maze test.The results of hematoxylin-eosin(HE)staining showed that the cells in the hippocampal tissue of the drug administration groups were closely arranged.Moreover,the drugs reduced the content of interleukin-6(IL-6,P<0.01)and tumor necrosis factor-α(TNF-α)in the hippocampal tissue,which were more obvious in the BBPR group(P<0.05).After screening,15 potential biomarkers

关 键 词：黑豆黄精 炆黄精 UPLC-Q/TOF-MS 代谢组学 阿尔茨海默病 

分 类 号：R285.5[医药卫生—中药学]