湖南中医药大学学报2026，Vol.46Issue(1)：51-61,11.DOI:10.3969/j.issn.1674-070X.2026.01.007
基于中医方证代谢组学研究六味地黄丸治疗孤独症谱系障碍肝肾不足证的物质基础
On the material basis of Liuwei Dihuang Pill in treating autism spectrum disorder with liver-kidney deficiency pattern based on TCM formula-pattern metabolomics
摘要
Abstract
Objective To investigate the pharmacodynamic material basis of Liuwei Dihuang Pill(LWDHP)on autism spectrum disorder(ASD)with liver-kidney deficiency pattern based on the strategy of TCM pattern-related metabonomics.Methods An ASD offspring rat model was established by intraperitoneal injection of sodium valproate(VPA)during pregnancy.Successfully modeled offspring rats were randomly divided into model group(n=8)and LWDHP group(n=8),with additional blank control group(n=8)and blank medication control group(n=5).Every morning,the LWDHP and the blank medication groups were gavaged with LWDHP suspension(3.4 g/kg),while the model and the blank control groups received an equal volume of distilled water.Every afternoon,the model and the LWDHP groups were gavaged with levothyroxine sodium(150 μg/kg),followed by a 5-minute tail clamp at approximately 2 cm from the tail tip as an emotional stimulation to establish a liver-kidney deficiency pattern.Both gavage and tail-clamp were performed once daily for 14 consecutive days.After treatment,the three-chamber social test was used to evaluate social behavior of offspring rats.HE staining and Nissl staining were performed to observe histopathological changes in the prefrontal cortex.Ultra performance liquid chromatography-quadrupole-time of flight-tandem mass spectrometry(UPLC-Q-TOF-MS)combined with multivariate statistical analysis was employed to identify differential metabolites,and pathway enrichment analysis was conducted using the MetaboAnalyst 5.0 platform.UPLC-Q-TOF-MS combined with the extracted ion chromatogram(EIC)function and mass spectrometry comparison was used to identify the brain-penetrating components of LWDHP.Molecular docking between the brain-penetrating components of LWDHP and key enzymes of differential metabolites was performed using AutoDock Vina software.Results The three-chamber social test results showed that in the sociability phase,compared with the blank control group,offspring rats in the model group spent significantly less time interacting with stranger rat one(P<0.01);compared with the model group,the LWDHP group spent more time interacting with stranger rat one(P<0.05).In the social novelty phase,compared with the blank control group,the model group spent significantly less time interacting with stranger rat two(P<0.01);compared with the model group,the LWDHP group spent more time interacting with stranger rat two(P<0.05).HE and Nissl staining results showed neuropathological damage in the prefrontal cortex of the model group compared with the blank control group,while this damage in the LWDHP group was alleviated compared to the model group.Metabolomic results indicated that the pathological state of ASD with liver-kidney deficiency pattern significantly affected the in vivo metabolic processes of LWDHP.Multivariate statistical analysis identified a total of 42 differential metabolites,among which purine,2-(2-aminoethyl)pyridine,tetradecylamine,butenoylcarnitine,sphingosine,pyridoxol(vitamin B6),and α-linolenic acid were identified as key differential metabolites in the treatment of ASD with liver-kidney deficiency pattern by LWDHP,involving sphingolipid metabolism,vitamin B6 metabolism,linolenic acid metabolism,and porphyrin metabolic pathways.Analysis of brain-penetrating components identified nine components of LWDHP,including betaine,2-furoic acid,adenosine,paeonol,piperlonguminine,dioscin,oleanolic acid,linoleic acid,and palmitic acid.Molecular docking results showed that key enzymes of the differential metabolites including adenylosuccinate lyase(ADSL),phosphoribosyl pyrophosphate synthetase(PRPS),fatty acid amide hydrolase(FAAH),amidase(Acy),carnitine palmitoyltransferase 1(CPT1),sphingosine kinase 2(SphK2),pyridoxal kinase(PDXK),and fatty acid desaturase 2(FADS2)exhibited the strongest binding energy with dioscin and oleanolic acid.The binding energy of ADSL,Acy,CPT1,PDXK,and FADS2 with dioscin,and of CPT1 and PDXK with oleanolic acid were all less than-10 kcal·mol-1.Conclusion The mechanism of action of LWDHP on ASD with liver-kidney deficiency pattern may be related to the regulation of sphingolipid metabolism,vitamin B6 metabolism,linolenic acid metabolism,and porphyrin metabolism pathways.Its material basis may involve betaine,2-furoic acid,adenosine,paeonol,piperlonguminine,dioscin,oleanolic acid,linoleic acid,and palmitic acid.This study provides a theoretical foundation for optimizing the preparation process,establishing quality standards,and developing innovative drugs for LWDHP.关键词
孤独症谱系障碍/六味地黄丸/肝肾不足证/方证代谢组学/分子对接Key words
autism spectrum disorder/Liuwei Dihuang Pill/liver-kidney deficiency pattern/formula-pattern metabolomics/molecular docking分类
医药卫生引用本文复制引用
裴方妤,吴吉,贺思雨,雷倩,赵莎,张涤,朱沁泉..基于中医方证代谢组学研究六味地黄丸治疗孤独症谱系障碍肝肾不足证的物质基础[J].湖南中医药大学学报,2026,46(1):51-61,11.基金项目
湖南省自然科学基金项目(2024JJ8233) （2024JJ8233）
湖南省中医药科研计划项目(A2023036) （A2023036）
湖南省卫生健康高层次人才项目(20230448) （20230448）
湖南中医药大学院校联合基金项目(2025XYLH038) （2025XYLH038）
2024年湖南省大学生创新创业训练计划项目(S202410541157). （S202410541157）
