基于UHPLC-Q-Exactive Orbitrap MS和分子网络技术研究补肾活血方在大鼠体内的吸收原型和代谢产物  

Profiling Absorbed Prototypes and Metabolites of Bushen Huoxue Prescription(补肾活血方):A Comprehensive UHPLC-Q-Exactive-MS/MS Study with Molecular Network Analysis

作　　者：王宇 匡微 黄宇霞 谢孟君 张梅[1] WANG Yu;KUANG Wei;HUANG Yuxia;XIE Mengjun;ZHANG Mei(State Key Laboratory of Southwestern Chinese Medicine Resources,School of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,Sichuan,China)

机构地区：[1]成都中医药大学药学院西南特色中药资源国家重点实验室,四川成都611137

出　　处：《中华中医药学刊》2025年第2期127-136,I0029-I0030,共12页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(82174273,81774202,82305323)。

摘　　要：目的补肾活血方由地黄、丹参、人参及葛根组成,目前已证实对糖尿病性视网膜病变有良好的临床疗效。该研究旨在分析此方在大鼠体内的吸收原型和代谢产物,以及这些成分在大鼠体内的分布差异。方法SD大鼠灌胃补肾活血方后,收集不同时间点的大鼠血浆、肝、心、脾、肺、肾、肠、胃、眼、尿液及粪便,采用超高效液相色谱-四极杆-静电场轨道阱高分辨质谱(UHPLC-Q-Exactive Orbitrap MS)采集数据,通过数据库匹配及比对相关文献中的相对分子质量、离子碎片等信息鉴定成分,并结合分子网络技术补充代谢产物。结果在大鼠体内共鉴定到75个吸收原型及123个代谢成分,包括苯乙醇苷类、丹参酮类、人参皂苷类、异黄酮类、环烯醚萜类、丹酚酸类等,主要集中于肝和肾中。苯乙醇苷类成分来源于地黄,其生物利用率低,易经过生物转化形成甲基化、葡萄糖醛酸化等代谢物,更多分布于肾中;丹参酮类及异黄酮类原型成分分别来源于丹参和葛根,在大鼠血浆和各组织中易被检测到,其代谢途径以羟基化、甲基化及葡萄糖醛酸化为主,产物易分布于血浆及肝肾组织中。人参皂苷类化合物在大鼠体内易被胃肠道水解为原人参二醇及原人参三醇,经脱氢、脱氧等I相代谢后,产物更多分布于脾。结论该研究初步阐明了补肾活血方原型成分及代谢产物的吸收、分布情况,为阐释补肾活血方及地黄、丹参、人参及葛根治疗糖尿病性视网膜病变的药效物质基础提供了科学依据。Objective Bushen Huoxue Prescription(补肾活血方,BHP),a traditional Chinese medicine formula,is used for the treatment of diabetic retinopathy(DR).It is composed of four herbal ingredients.However,the absorbed prototypes and metabolites of BHP have not been extensively studied.Methods The plasma,liver,heart,spleen,lung,kidney,intestine,stomach,eye,urine and feces of SD rats at different time points were collected after intragastric administration of BHP.The absorbed prototypes and metabolites of BHP in rats were characterized using the UHPLC-Q-Exactive-MS/MS strategy.The metabolic networks in rats after oral administration of BHP were constructed by molecular network.Results Seventy-five absorbed prototypes and 123 metabolites were identified in rats belonging to different groups in BHP,including iridoids,phenylethanol glycosides,tanshinones,ginsenosides,isoflavonoids and other types of compounds.Phenylethanoid glycosides were derived from Dihuang(Rehmanniae Radix),which had low bioavailability and were easy to be biotransformed into metabolites such as methylation and glucuronidation,and were more distributed in the kidney.The prototype components of tanshinones and isoflavones were derived from Danshen(Salviae Miltiorrhizae Radix et Rhizoma)and Gegen(Puerariae Lobatae Radix)respectively,which were easily detected in rat plasma and tissues.The metabolic pathways were mainly hydroxylation,methylation and glucuronidation,and the products were easily distributed in plasma and liver and kidney tissues.Ginsenosides were easily hydrolyzed into protopanaxadiol and protopanaxatriol in the gastrointestinal tract of rats.After phase I metabolism such as dehydrogenation and deoxidation,the products were more distributed in the spleen.Conclusion These findings could reveal the mechanism of action of BHP in DR treatment and provide a method for the discovery of metabolites and metabolic pathways.

关 键 词：补肾活血方 吸收原型 代谢产物 组织分布 分子网络技术 超高效液相色谱-四极杆-静电场轨道阱高分辨质谱 

分 类 号：R282.71[医药卫生—中药学]