关白附炮制前后对缺血性中风沙鼠药效学和代谢组学的影响  

Effect of Guanbaifu(Radix Aconiti Coreani)Before and After Processing on the Metabolomics of Gerbils with Ischemic Stroke

作　　者：曾琬婷 周丽婷 贾茹 刘月 马浩文 才谦[1] 曲扬[1] ZENG Wanting;ZHOU Liting;JIA Ru;LIU Yue;MA Haowen;CAI Qian;QU Yang(College of Pharmacy,Liaoning University of Traditional Chinese Medicine,Dalian 116600,Liaoning,China)

机构地区：[1]辽宁中医药大学药学院,辽宁大连116600

出　　处：《辽宁中医药大学学报》2024年第4期55-63,共9页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：辽宁省“兴辽英才”计划科技创新领军人才(特聘教授)项目(XLYC2002004)。

摘　　要：目的研究关白附生品及其炮制品对沙鼠缺血性脑中风的保护作用及对尿液代谢物的影响。方法将36只沙鼠随机分成空白组(CG)、模型组(MG)、阳性药组(PG)、生关白附组(RA)、蒸关白附组(SA)和姜矾制关白附组(GA),每组6只,除空白组沙鼠外,其余各组采用右侧单侧结扎沙鼠颈动脉造成中风模型。造模成功后,关白附给药组分别灌胃对应的饮片全粉混悬液(0.586 mg/g),阳性药组灌胃脑络通颗粒混悬液(0.390 mg/g),空白组灌胃生理盐水,共给药14 d。分别采用Longa评分法、氯化三苯基四氮唑(TTC)染色法、酶联免疫法检测关白附生品及其炮制品对神经功能学评分、脑组织梗死面积、氧化应激水平、炎症因子表达;采用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱法(UHPLC-Q-Orbitrap-MS)分析空白组、模型组、阳性药组、生关白附组、蒸关白附组和姜矾制关白附组沙鼠尿液样品;采用多元统计方法对数据进行处理,筛选差异代谢物;采用京都基因与基因组百科全书(KEGG)数据库结合MetaboAnalyst 5.0软件对筛选出的差异代谢物进行代谢通路分析。结果关白附及其炮制品可显著改善脑组织梗死面积;明显降低脑组织和血清中丙二醛(MDA),白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)活性(P<0.05),明显升高超氧化物歧化酶(SOD)、谷胱甘肽(GSH)、一氧化氮(NO)活性(P<0.05);代谢组学研究发现L-脯氨酸(L-Proline)、核黄素(Riboflavin)、白三烯D4(Leukotriene D4)、7-甲基黄嘌呤(7-Methylxanthine)等112种差异代谢物可作为缺血性脑中风潜在生物标志物,涉及核黄素代谢、嘧啶代谢、嘌呤代谢等14条代谢通路。生关白附回调了其中的核黄素代谢、嘧啶代谢等4条代谢通路;蒸关白附回调了其中的核黄素代谢、嘧啶代谢、精氨酸和脯氨酸代谢等5条代谢通路;姜矾制关白附回调了其中的咖啡因代谢和乙醛和二羧酸代谢2条代谢通路Objective To investigate the effects of raw Guanbaifu(Radix Aconiti Coreani),and its processed products on the urinary metabolites of gerbils with ischemic stroke using non-targeted metabolomics techniques to elucidate the mechanism of anti-stroke effects.Methods Thirty-six gerbils were randomly divided into a control group(CG),a model group(MG),a positive drug group(PG),a raw Guanbaifu(Radix Aconiti Coreani)group(RA),a steamed Guanbaifu(Radix Aconiti Coreani)group(SA)and a Guanbaifu(Radix Aconiti Coreani)processed with ginger and alum group(GA),with six gerbils in each group,except for the CG.After successful modeling,the RA,SA and GA were gavaged with the corresponding herbal whole powder suspension(0.586 mg/g),PG was gavaged with suspension of Naoluotong Granules(脑络通颗粒,0.390 mg/g)and the CG was gavaged with saline for 14 days.Longa score,triphenyltetrazolium chloride(TTC)staining,and enzyme-linked immunoassay were used to detect the effects of Guanbaifu(Radix Aconiti Coreani)and their processed products on neurological function score,brain tissue infarct volume,oxidative stress level,and inflammatory factor expression,respectively.The urine samples of gerbils in the CG,MG,PG,RA,SA and GA were analyzed by ultra performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry(UHPLC-Q-Orbitrap-MS);the data were processed by multivariate statistical methods to screen the differential metabolites;the Kyoto Encyclopedia of Genes and Genomes(KEGG)database and MetaboAnalyst 5.0 software were used to analyze the metabolic pathways of the screened differential metabolites.Results Guanbaifu(Radix Aconiti Coreani)and its processed products significantly improved the infarcted area of brain tissue;significantly reduced malondialdehyde(MDA),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)activities in brain tissue and serum(P<0.05),and significantly increased superoxide dismutase(SOD),glutathione(GSH)and nitric oxide(NO)activities(P<0.05).The metabolomic study iden

关 键 词：关白附 炮制品 沙鼠 缺血性脑中风 代谢组学 多元统计分析 生物标志物 

分 类 号：R285.5[医药卫生—中药学]