基于血浆代谢组学与谱效相关的柴胡和白芍醋炙及配伍前后抗肝纤维化作用机制和物质基础研究  

Integration of plasma metabolomics and spectrum-effect correlation for profiling the mechanism and pharmacodynamic substances on attenuating hepatic fibrosis of Bupleuri Radix and Paeoniae Radix Alba before and after vinegar-processing and compatibility

作　　者：刘鑫 于慧 宋健涛[1,2] 陈忆菁 张巧 蔡皓 曹岗[3] 朱慧[1,2] 段煜[1,2] 裴科[4] LIU Xin;YU Hui;SONG Jian-tao;CHEN Yi-jing;ZHANG Qiao;CAI Hao;CAO Gang;ZHU Hui;DUAN Yu;PEI Ke(School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China;Engineering Center of State Ministry of Education for Standardization of Chinese Medicine Processing,Nanjing University of Chinese Medicine,Nanjing 210023,China;School of Pharmacy,Zhejiang Chinese Medical University,Hangzhou 310053,China;School of Chinese Medicine and Food Engineering,Shanxi University of Chinese Medicine,Jinzhong 030619,China)

机构地区：[1]南京中医药大学药学院,江苏南京210023 [2]南京中医药大学国家教育部中药炮制规范化及标准化工程研究中心,江苏南京210023 [3]浙江中医药大学药学院,浙江杭州310053 [4]山西中医药大学中药与食品工程学院,山西晋中030619

出　　处：《药学学报》2023年第7期1790-1801,共12页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金面上项目(81673600,81973481);江苏省研究生科研创新计划(KYCX21_1778,KYCX21_1790).

摘　　要：本研究利用UHPLC-Q-TOF-MS/MS技术结合血浆代谢组学方法分析柴胡和白芍醋炙及配伍前后抗肝纤维化的作用机制,并通过谱效相关方法筛选潜在的药效物质。通过皮下注射CCl4橄榄油溶液复制肝纤维化大鼠模型,分别灌胃给予柴胡、醋柴胡、白芍、醋白芍、“柴胡-白芍”药对及“醋柴胡-醋白芍”药对水煎液,持续给药六周后收集血样和组织样品用于药效指标检测及血浆代谢组学分析。本实验获得南京中医药大学实验动物伦理委员会批准(批准号:202103A002)。药效学结果表明,各给药组均能降低丙氨酸氨基转移酶(ALT,P<0.01)、天门冬氨酸氨基转移酶(AST,P<0.01)和羟脯氨酸(HYP,P<0.01)水平,显著提升谷胱甘肽过氧化物酶(GSH-Px,P<0.05)水平。通过UHPLC-Q-TOF-MS/MS技术对内源性代谢物进行分析,在正、负离子模式下共鉴定出20种与肝纤维化发病机制相关的差异性代谢物,主要涉及到视黄醇代谢、甘油磷脂代谢、甘油酯代谢、脂肪酸生物合成和花生四烯酸代谢5条代谢通路。同时,引入成分矫正率的概念用以评估各给药组对差异性代谢物的回调作用,各给药组对其中10种差异性代谢物均有矫正作用,且醋炙品组的矫正作用优于生品组,药对组的矫正作用优于单味药组,其中,以“醋柴胡-醋白芍”药对组的回调作用最佳,进一步验证了醋炙及配伍对药效的提升。对入血成分及代谢产物进行鉴定后,采用偏最小二乘法及VIP分析,并借助谱效相关确定了与ALT、AST、HYP和GSH-Px四个经典药效指标具有较高关联度的38种成分作为柴胡和白芍醋炙及配伍前后抗肝纤维化作用的药效标志物。研究结果不仅可为阐明柴胡和白芍醋炙及配伍治疗肝纤维化的药效物质及作用机制奠定基础,为阐明炮制与配伍的科学内涵提供理论依据,而且可进一步为柴胡和白芍的临床用药设计与开发提供参考。Plasma metabolomics based on UHPLC-Q-TOF-MS/MS technique was developed for profiling the mechanism on attenuating hepatic fibrosis of Bupleuri Radix(BR)and Paeoniae Radix Alba(PRA)before and after vinegar-processing and compatibility,and to screen potential pharmacodynamic substances by spectrumeffect correlation method in this study.Firstly,SD rats with CCl4-induced hepatic fibrosis were used as an in vivo model.The blood and tissue samples were collected for the analyses of pharmacodynamic indexes and plasma metabolomics after six weeks’administration of BR,vinegar-processed BR(VPBR),PRA,vinegar-processed PRA(VPPRA),BR-PRA herb-pair,and VPBR-VPPRA herb-pair.The experiment was approved by the experimental animal ethics committee from Nanjing University of Chinese Medicine(No.202103A002).The results of pharmacodynamics indicated that the levels of alanine aminotransferase(ALT,P<0.01),aspartate aminotransferase(AST,P<0.01),and hydroxyproline(HYP,P<0.01)were decreased significantly,while the level of glutathione peroxidase(GSH-Px,P<0.05)was increased obviously after administration of all treatment groups.Next,UHPLC-QTOF-MS/MS was performed to characterize the endogenous metabolites.A total of 20 differential endogenous metabolites related to the pathogenesis of hepatic fibrosis were identified in positive and negative ion modes,mainly involving five metabolic pathways of retinol metabolism,glycerol phospholipid metabolism,glyceride metabolism,fatty acid biosynthesis,and arachidonic acid metabolism.Meanwhile,a concept named correction rate was introduced to evaluate the back-regulation effects of all treatment groups on differential metabolites,and 10 differential metabolites were corrected by all treatment groups.The correction effects of the vinegar-processed herb groups were better than those of the crude ones,and the correction effects of the herb-pair groups were better than those of the single ones.Interestingly,the best correction effect was found in the VPBR-VPPRA herb-pair group,which further verified the

关 键 词：柴胡 白芍 醋炙 药对 入血成分 代谢产物 谱效相关 

分 类 号：R917[医药卫生—药物分析学]