| Traditional Chinese medicine exerts its therapeutic effect as a whole by its synergistic effects with various characteristic components.Chinese medicine is widely used in China and other Asian countries for disease prevention,diagnosis,treatment or regulation of human functions,and has gradually become an effective complementary therapy in the Western countries.Under the guidance of basic theories of Chinese medicine,Chinese medicine is a unique medical system that has the distinctive features of multi-target effect and the pursuit of overall therapeutic effect.It consists of a variety of major or minor components that can regulate the body’s balance and homeostasis as a whole.Citrus is a phytomedicine with homology of medicine and food.Because it contains a large number of polyphenols,it has an important therapeutic effect on human health,and citrus fruits and fruit juices are also found all over the world.The content and distribution of bioactive phenolic compounds in the Citrus are very different with the fruit’s maturation process.Zhishi,Zhiqiao,Qingpi and Chenpi are used as traditional Chinese medicines to regulate blood and strengthen spleen and stomach since ancient times,and its actual clinical application in China is diverse.In this study,a research strategy based on pseudotargeted metabolomics techniques that can be used to explore reliable material bases for the efficacy of Chinese medicines is proposed.Pseudotrgeted metabolomics combines the advantages of both targeted and non-targeted technologies.Targeted metabolomics based on the optimal conditions of the RRLC-QqQ-MS method for in vivo quantification plasma pharmacochemistry of characteristic prototyping components Untargeted metabolomics using UHPLC-QE Orbitrap HRMS with better specificity and resolution was used to identify untargeted metabolites in complex plasma matrices.Finally,32 prototype metabolites in the plasma,66 biotransformed metabolites after oral administration of the standard extracts of the four labeled citrus herbs were quantitatively determined.Systemically describes the absorption and metabolism of complex Chinese herbs in the body.During our previous study,we discovered the potential impact of Citrus on the cytochrome P450 family of enzymes,suggesting a possible resistance to the toxicity of the overdose of acetaminophen(APAP).APAP is one of the most widely used drugs and is a widely used drug to treat pain and fever.However,overdose can always cause significant liver damage and even life-threatening.In our previous study,as a representative Chinese herbal medicine that exerted its therapeutic effect with multiple components,it was shown to inhibit the toxic metabolic activation of APAP.In a further study,we tested the protective effect of Zhishi on APAP-induced liver injury in rats and BRL-3A cells.After seven-day pretreatment of Zhishi,the serum levels of ALT and AST decreased,and also,the pathological lesions of the liver were reduced.This indicated that the Zhishi had a significant protective effect on APAP-induced liver injury and APAP-induced death of BRL-3A cells.Then,a metabolomics strategy was used to study the prevention of APAP-induced liver injury by regulating hepatic metabolic disorders.Based on multivariate statistical analysis such as PCA and OPLS-DA,significant differences in metabolites between the APAP treated group and the control group were determined.Finally,potential biomarkers of 13 amino acids,13 phosphatidylcholines(PC),11 phosphatidylethanolamines(PE)and 8 other compounds were screened and identified.And the metabolic pathways most involved in the treatment of diseases:glycerophospholipid metabolism,fatty acid biosynthesis,and glycerol lipid metabolism.In addition,we have found that Zhishi against APAP-induced hepatotoxicity by modulating PUMA,AMPK-SIRT1,and JNK signaling pathways to blocking apoptosis and promoting liver regeneration.In summary,the results of our research show that based on the pseudotargeted metabolomics technology,the transitional components and possible pharmacodynamic material basis of the Citrus are demonstrated,also Provides new research methods to find and study the pharmacodynamic material basis of other Chinese medicines.At the same time,Zhishi protects the liver against liver toxicity caused by excessive APAP by inhibiting p53 up-regulation of apoptotic regulators and reversing hepatic lipid metabolism disorder,which gives us insights into the potential treatment effect of Citrus TCM in the field of drug-induced organ injury for the future.And also contribute to the theoretical basis of drug development and drug combination. |
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