ETHNOPHARMACOLOGICAL RELEVANCE:Zingiberis rhizoma recens-/wine-/euodiae fructus-processed Coptidis Rhizoma (CR, zCR/wCR/eCR) are the commonly used processed products of CR in clinic. After being processed with different excipients, the efficacy of CR will change accordingly. I.e., wCR could resolve excessive heat of the upper energizer, zCR could eliminate gastric heat and harmonize the stomach, eCR could smooth the liver and harmonize the stomach. However, the underlying mechanisms were still unclear.AIM OF THE STUDY:To further verify the differential efficacy of the three processed CR products and compare the mechanisms on gastric ulcer.MATERIAL AND METHODS:First, a GU model, whose onset is closely related to the heat in stomach and the disharmony between liver and stomach, was established, and the therapeutic effects of zCR/wCR/eCR/CR were evaluated by pathologic observation and measurement of cytokine levels. Second, metabolomics analysis and network pharmacology were conducted to reveal the differential intervening mechanism of zCR/eCR on GU. Third, the predicted mechanisms from metabolomics analysis and network pharmacology were validated using western blotting, flow cytometry and immunofluorescence.RESULTS:zCR/wCR/eCR/CR could alleviate the pathologic damage to varying degrees. In metabolomics research, fewer metabolic pathways were enriched in serum samples, and most of them were also present in the results of gastric tissue samples. The gastroprotective, anti-inflammatory, antioxidant, and anti-apoptotic effects of zCR/wCR/eCR/CR might be due to their interference on histidine, arachidonic acid, and glycerophospholipids metabolism. Quantitative results indicated that zCR/eCR had a better therapeutic effect than wCR/CR in treating GU. A comprehensive analysis of metabolomics and network pharmacology revealed that zCR and eCR exerted anti-GU effects via intervening in five core targets, including AKT, TNF, IL6, IL1B and PPARG. In the validation experiment, zCR/eCR could significantly reverse the abnormal expression of proteins related to apoptosis, inflammation, oxidative stress, gastric function, as well as the PI3K/AKT signaling pathways.CONCLUSION:zCR and eCR could offer gastroprotective benefits by resisting inflammation and apoptosis, inhibiting gastric-acid secretion, as well as strengthening gastric mucosal defense and antioxidant capacity. Integrating network pharmacology and metabolomics analysis could reveal the acting mechanism of drugs and promote the development of medications to counteract GU.

Iridoid components have been reported to have significant neuroprotective effects. However, it is not yet clear whether the efficacy and mechanisms of iridoid components with similar structures are also similar. This study aimed to compare the neuroprotective effects and mechanisms of eight iridoid components (catalpol (CAT), genipin (GE), geniposide (GEN), geniposidic acid (GPA), aucubin (AU), ajugol (AJU), rehmannioside C (RC), and rehmannioside D (RD)) based on corticosterone (CORT)-induced injury in PC12 cells. PC12 cells were randomly divided into a normal control group (NC), model group (M), positive drug group (FLX), and eight iridoid administration groups. Firstly, PC12 cells were induced with CORT to simulate neuronal injury. Then, the MTT method and flow cytometry were applied to evaluate the protective effects of eight iridoid components on PC12 cell damage. Thirdly, a cell metabolomics study based on ultra-performance liquid chromatography–quadrupole–time-of-flight mass spectrometry (UPLC-Q/TOF-MS) was performed to explore changes in relevant biomarkers and metabolic pathways following the intervention of administration. The MTT assay and flow cytometry analysis showed that the eight iridoid components can improve cell viability, inhibit cell apoptosis, reduce intracellular ROS levels, and elevate MMP levels. In the PCA score plots, the sample points of the treatment groups showed a trend towards approaching the NC group. Among them, AU, AJU, and RC had a weaker effect. There were 38 metabolites (19 metabolites each in positive and negative ion modes, respectively) identified as potential biomarkers during the experiment, among which 23 metabolites were common biomarkers of the eight iridoid groups. Pathway enrichment analysis revealed that the eight iridoid components regulated the metabolism mainly in relation to D-glutamine and D-glutamate metabolism, arginine biosynthesis, the TCA cycle, purine metabolism, and glutathione metabolism. In conclusion, the eight iridoid components could reverse an imbalanced metabolic state by regulating amino acid neurotransmitters, interfering with amino acid metabolism and energy metabolism, and harmonizing the level of oxidized substances to exhibit neuroprotective effects.

Rhizoma coptidis (CR) is traditionally used for treating gastrointestinal diseases. Wine-processed CR (wCR), zingiber-processed CR (zCR), and evodia-processed CR (eCR) are its major processed products. However, the related study of their specific mechanisms is very limited, and they need to be further clarified. The aim of this study is to compare the intervening mechanism of wCR/zCR/eCR on rats via faecal metabolomics and 16S rDNA gene sequencing analysis. First, faecal samples were collected from the control and CR/wCR/zCR/eCR groups. Then, a metabolomics analysis was performed using UHPLC-Q/TOF-MS to obtain the metabolic profile and significantly altered metabolites. The 16S rDNA gene sequencing analysis was carried out to analyze the composition of gut microbiota and screen out the significantly altered microbiota at the genus level. Finally, a pathway enrichment analysis of the significantly altered metabolites via the KEGG database and a functional prediction of relevant gut microbes based on PICRUSt2 software were performed in combination. Together with the correlation analysis between metabolites and gut microbiota, the potential intervening mechanism of wCR/zCR/eCR was explored. The results suggested that wCR played a good role in maintaining immune homeostasis, promoting glycolysis, and reducing cholesterol; zCR had a better effect on protecting the integrity of the intestinal mucus barrier, preventing gastric ulcers, and reducing body cholesterol; eCR was good at protecting the integrity of the intestinal mucus barrier and promoting glycolysis. This study scientifically elucidated the intervening mechanism of wCR/zCR/eCR from the perspective of faecal metabolites and gut microbiota, providing a new insight into the processing mechanism research of Chinese herbs.

Ephedra herba is a conventional Chinese medicine to treat cold, fever, asthma, edema, and lung diseases in the clinic. At present, most pharmacokinetic studies focus on the pharmacokinetic process of alkaloids in normal animals. However, the non-alkaloid components are also active. In addition, the pharmacokinetic studies under pathological state make more sense for clarifying the material basis of efficacy. In this study, a sensitive and rapid ultra-high-performance-tandem mass spectrometry method was developed and applied to determine nine bioactive components (ephedrine, pseudoephedrine, methylephedrine, (+)-catechin, epicatechin, vitexin, vicenin-2, cinnamic acid, and ferulic acid) in normal, common cold and nephrotic syndrome rats after the oral administration of Ephedra herba. Compared to the normal group, except for ferulic acid, the exposure levels of the other eight components were significantly increased and the plasma clearance clearly declined in common cold rats. Similarly, the exposure levels of seven components other than cinnamic acid and ferulic acid were also significantly augmented and the plasma clearance decreased significantly in nephrotic syndrome rats. In brief, the pathological conditions of the common cold and nephrotic syndrome could lead to alterations in the pharmacokinetics profiles of the nine components, which provide a reference for further exploration of the pharmacodynamics basis of Ephedra herba.

Abstract Background According to the theory of TCM, ascending medicine has the effects of elevating yang, raising drooping, and inducing sweating. Descending medicine has the effects of relieving dyspnea and hiccup, stopping reflux, and calming nerves. In conclusion, the ascending/descending properties of TCM serve as a guide for the use of TCMs in clinical practice. However, at present, there are few studies on the ascending/descending properties of TCM. This study aimed to compare the effects of 6 ascending and 8 descending medicine on the main organs (heart, liver, lung, spleen, and kidney) of normal rats, characterizing the ascending and descending properties. Methods The orgain samples of all experiment group were analyzed based on ultra performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-Q/TOFMS). Then, to character the ascending and descending properties, the effects of 6 ascending and 8 descending medicine on the main organs (heart, liver, lung, spleen, and kidney) of normal rats were compared. Results The systemic metabolomics results indicated that linoleic acid metabolism and arginine and proline metabolism were the major differential pathways affected by ascending/descending medicine in the heart; nicotinic acid and nicotinamide metabolism, glutamic acid and glutathione metabolism, and L-carnitine metabolism were the major differential pathways affected by ascending/descending medicine in the liver; tryptophan metabolism, phenylalanine metabolism, and pyroglutamic acid and glutamic acid metabolism were the major differential pathways affected by ascending/descending medicine in the lung; tryptophan metabolism, phenylalanine metabolism, and glutamic acid metabolism were the major differential pathways affected by ascending/descending medicine in the kidney; and glutamic acid and glutamine metabolism were the major differential pathway affected by ascending/descending medicine in the spleen. Conclusions Ascending medicine exhibited positive effects on nerve stimulation, immunity, and reproductive function by promoting energy metabolism in the heart and liver, whereas descending medicine had the opposite effect. Descending medicine exhibited a positive effect on diuresis by promoting blood circulation, whereas ascending medicine had the opposite effect. Both descending medicine and ascending medicine intervened with pulmonary inflammation, but the underlying mechanisms may be different. This study demonstrated that systemic metabolomics technology can be used for the characterization of traditional Chinese medicine properties.

Background: Descurainia sophia seeds (DS) is a herbal medicine in traditional Chinese medicine (TCM) for treating lung diseases. We aimed to evaluate the therapeutic effect of DS and five of its fractions upon pulmonary edema (PE) through metabolomics analysis (MA) of urine and serum samples of rats. Methods: A PE model was established by intrathoracic injection of carrageenan. Rats were pretreated with DS extract or its five fractions (polysaccharides (DS-Pol); oligosaccharides (DS-Oli); flavonoid glycosides (DS-FG); flavonoid aglycone (DS-FA); fat oil fraction (DS-FO)) for seven consecutive days. Forty-eight hours after carrageenan injection, lung tissues were subjected to histopathology. MA of urine and serum was done by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry, respectively. Principal component analysis and orthogonal partial least squares-discriminant analysis were operated for the MA of rats and potential biomarkers related to treatment. Heatmaps and metabolic networks were constructed to explore how DS and its five fractions act against PE. Results: DS and its five fractions could all attenuate pathologic lung injury to different degrees, and DS-Oli, DS-FG, and DS-FO had a more potent effect compared with DS-Pol and DS-FA. DS-Oli, DS-FG, DS-FA, and DS-FO could regulate the metabolic profiles of PE rats, but DS-Pol was less potent. According to MA, the five fractions could improve PE to some degree due to their anti-inflammatory, immunoregulatory, and renoprotective activities by mediating the metabolism of taurine, tryptophan, and arachidonic acid. However, DS-Oli, DS-FG, and DS-FO had more important roles in edema-fluid reabsorption, and reduction of vascular leakage through regulating the metabolism of phenylalanine, sphingolipid and bile acid. Finally, heatmaps and hierarchical clustering analysis indicated DS-Oli, DS-FG, and DS-FO to be more efficacious than DS-Pol or DS-FA against PE. The five fractions of DS had a synergistic effect on PE from different aspects, thereby constituting the entire efficacy of DS. DS-Oli, DS-FG, or DS-FO could be used as an alternative to DS. Conclusion: MA combined with use of DS and its fractions provided novel insights into the mechanism of action of TCM.

The efficacy of Rehmannia Radix changes after processing. However, the precise effect of processing on the properties of Rehmannia Radix is an intricate topic, as this effect cannot be explained by traditional methods. The purpose of this study was to investigate how processing methods influence the properties of Rehmannia Radix, as well as the changes in body function after administering dried Rehmannia Radix (RR) and processed Rehmannia Radix (PR) using a metabolomics approach. In addition, principal component analysis and orthogonal partial least-squares discriminant analysis models were generated using SIMCA-P 14.0 to evaluate the properties of RR and PR. Potential biomarkers were identified, and associated metabolic networks were established to clarify differences in the properties and efficacies of RR and PR. The results showed that RR and PR have cold and hot properties, respectively. RR can exert a hypolipidaemic effect by regulating nicotinate and nicotinamide metabolism. PR exerts a tonic effect and regulates the body's reproductive function through the regulation of alanine, aspartate and glutamate metabolism, arachidonic acid, pentose and glucuronate metabolism, respectively. Ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry-based metabolomics is a promising approach to determine the cold/hot properties of traditional Chinese medicine formulations.

Background: After being processed with different excipients, the clinical application of Coptidis Rhizoma (CR) is differentially investigated. However, the underlying mechanism and material basis are not clear, and there is a lack of attention to the collaborative working mode of herbal medicine during exploration. Purpose: To characterize the specific mechanism of wine/zingiberis rhizoma recens/euodiae fructus processed CR (wCR/zCR/eCR) and to investigate the role of excipients during processing. Methods: The multi-organ metabolomics approach was employed to explore the target organs of wCR/zCR/eCR and multiple pathways being triggered in each organ. The tissue distribution of CR and wCR/zCR/eCR com-ponents was compared to indicate the material basis of efficacy change after processing. Further, the network pharmacology study coupled with experimental validation was conducted to support metabolomic research and predicted active ingredients and core targets, and the molecular docking coupled with binding test was per-formed to identify the binding between active ingredient and core target. Results: The multi-organ metabolomics and network pharmacology study elucidated the intervening effect of wCR on heart/lung, zCR on stomach/colon, and eCR on liver/colon/stomach. Combined with molecular docking, binding test and tissue distribution studies, the specific mechanism was as follows: the wine made iso-quinoline alkaloids in CR more likely to accumulate in heart/lung, thus triggering the core targets of PTGS2, NOS2, ESR1 and SLC6A4 in heart/lung, and thereby highlighting the detoxifying and cardiopulmonary protective effect of wCR. The zingiberis rhizoma recens and euodiae fructus made organic acids in CR more likely to accumulate in stomach/colon and liver/colon/stomach respectively, thus triggering the core targets of ACTB, TNF and PRKCA in stomach/colon, the core targets of ACTB, TNF, PRKCA and GPT in stomach/colon/liver, and thereby high-lighting the improving effect of zCR/eCR on digestive function. Conclusion: Iso-quinoline alkaloids were the material basis of CR for anti-inflammation, and organic acids were mainly responsible for regulating gastrointestinal function. Due to the influence of excipients on the accumu-lation tendency of CR components, the differentially highlighted application of wCR/zCR/eCR was achieved.These findings propose a novel strategy for processing mechanism research.

Wine/zingiberis rhizoma recens/euodiae fructus processed Coptidis Rhizoma (wCR/zCR/eCR) are the major processed products of CR in clinic, and the role of CR is highlighted in different aspects after being processed with different excipients. To explore the mechanism and material basis for the highlighted efficacy of wCR/zCR/eCR, the metabolomics strategy was introduced to the comparative study between wCR/zCR/eCR and CR. Firstly, the metabolomics approach was applied to compare the chemical profiling and differential components between wCR/zCR/eCR and CR extract. Secondly, the rats were treated with CR/wCR/zCR/eCR extracts and a serum metabolomics approach was adopted to compare the metabolic profiling and significantly changed metabolites in CR/wCR/zCR/eCR groups, base on which the metabolic pathways were enriched, the metabolic network was constructed and the highlighted efficacy wCR/zCR/eCR was investigated. Lastly, the pathological and biochemical assessments (VIP, COX, HSL and HMGR) were implemented to validate the results inferred from metabolomics study. In chemical research, 23 differential components between wCR/zCR/eCR and CR extracts were identified. Thereinto, the content of alkaloids and organic acids decreased in wCR extract, the content of partial alkaloids and most organic acids increased in zCR extract, the content of alkaloids decreased, and partial organic acids increased in eCR extract. In serum metabolomics study, wCR had no outstanding effect, zCR played a more prominent role in resisting inflammation of gastrointestinal tissue by interfering with arachidonic acid metabolism, eCR exhibited the hottest drug property and the strongest effect on smoothing the liver and harmonizing the stomach by interfering with of bile acids biosynthesis. Based on the changes in chemical composition and efficacy before and after processing, as well as biochemical validation, it can be concluded that the above activity of zCR might be related to the increased alkaloids and organic acids in zCR extract, and the prominent role of eCR may be related to the increased organic acids in eCR extract. In brief, hot processing excipients could alleviate the cold property of CR, and different excipients have different effects on the chemical composition and efficacy mechanism. The present study fully reflects the advantage of metabolomics and provides guidance for the rational use of CR.

目的 基于HPLC-Q/TOF-MS代谢组学技术研究大黄水提物对正常大鼠的干预作用及相关机制.方法 将大鼠分为正常组和大黄水提物组,给药2周后收集各组大鼠尿液、血清样本,处理后进样分析,并对采集的数据进行多元统计分析、代谢网络富集、生物学功能解析.结果 尿液和血清样本的主成分分析得分图显示,大黄组与正常组均显著分离.经正交偏最小二乘判别分析,分别鉴定出生物标志物52个(尿液)、38(血清)个.代谢网络富集和生物学功能解析显示,这些生物标志物主要涉及酮体的合成与降解、谷氨酸和谷氨酰胺代谢、牛磺酸和亚牛磺酸代谢、淀粉和蔗糖代谢、丙氨酸、天冬氨酸和谷氨酰胺代谢、咖啡因代谢、三羧酸循环等代谢途径.结论 大黄可影响机体的脂质代谢、氨基酸代谢、神经递质以及肠道微生物等代谢途径,从而发挥其泻下通便、降血脂、改善胃肠道疾病等作用.

ETHNOPHARMACOLOGICAL RELEVANCE:Gleditsiae sinensis fructus Pills (GF) is a famous classical prescription, that is regularly combined with Jujubae fructus (JF) for the treatment of chronic bronchitis (CB) in the clinic. While the clinical efficacy of this combination prescription is clearly established, the active ingredients and molecular mechanisms remain unclear. AIM OF THE STUDY:To elucidate the mechanisms of action of Gleditsiae sinensis fructus Pills combined with Jujubae fructus (GF&JF) against CB based on network pharmacology and experimental verification. MATERIALS AND METHODS:The potential targets of GF&JF involved in therapeutic activity against CB were predicted based on network pharmacology and an "ingredients-targets" network constructed. The Metascape database was used for Module, GO functional and KEGG signaling pathway enrichment analyses of potential targets. Molecular docking was applied to simulate the binding activities of key candidate active ingredients to core targets. For experimental verification, a CB model was established through smoking and nasal cavity drip of lipopolysaccharide. Related inflammatory factors, including TNF-α, TGF-β, IL-6 and IL-8 in serum, and IL-4 IL-8, IFN-γ and IL-10 in bronchoalveolar lavage fluid (BALF), were detected using ELISA. Hematoxylin and eosin (H&E) and Masson staining were performed to observe pathological changes in lung and tracheal tissue. The expression of related proteins and mRNAs in the lung tissue were detected using immunohistochemistry (IHC), quantitative real-time PCR, and western blot. RESULTS:In network pharmacology, 36 common targets of GF&JF for CB were screened and the key targets and main signaling pathways identified. The active ingredients quercetin and stigmasterol in GF&JF had more targets for CB, which displayed good binding activity to IL-6, VEGFA, and EGFR, as established from molecular docking results. In vivo, GF&JF effectively inhibit the inflammatory response in CB mice and improved pathological changes in lung and tracheal tissue. In terms of the key proteins of the AGE-RAGE signaling pathway, GF&JF induced significant down-regulation of IL-6, ICAM-1, VCAM-1, EGFR, CASPASE-3, AGEs and RAGE proteins in lung tissue as well as mRNA expression of IL-6, ICAM-1, VCAM-1, EGFR, AGEs and RAGE. CONCLUSIONS:The GF&JF combination exerts a good therapeutic effect in CB model mice, which may be attributed to inhibition of the inflammatory response as well as regulation on the expression of AGE-RAGE signaling pathway. In addition, quercetin and stigmasterol appear to be the main active ingredients of GF&JF in the treatment of CB.

Background:The prognosis is poor when acute pancreatitis (AP) progresses to sepsis; therefore, it is necessary to accurately predict the probability of sepsis and develop a personalized treatment plan to reduce the disease burden of AP patients. Methods:A total of 1295 patients with AP and 43 variables were extracted from the Medical Information Mart for Intensive Care (MIMIC) IV database. The included patients were randomly assigned to the training set and to the validation set at a ratio of 7 : 3. The chi-square test or Fisher's exact test was used to test the distribution of categorical variables, and Student's t-test was used for continuous variables. Multivariate logistic regression was used to establish a prognostic model for predicting the occurrence of sepsis in AP patients. The indicators to verify the overall performance of the model included the area under the receiver operating characteristic curve (AUC), calibration curves, the net reclassification improvement (NRI), the integrated discrimination improvement (IDI), and a decision curve analysis (DCA). Results:The multifactor analysis results showed that temperature, phosphate, calcium, lactate, the mean blood pressure (MBP), urinary output, Glasgow Coma Scale (GCS), Charlson Comorbidity Index (CCI), sodium, platelet count, and albumin were independent risk factors. All of the indicators proved that the prediction performance and clinical profitability of the newly established nomogram were better than those of other common indicators (including SIRS, BISAP, SOFA, and qSOFA). Conclusions:The new risk-prediction system that was established in this research can accurately predict the probability of sepsis in patients with acute pancreatitis, and this helps clinicians formulate personalized treatment plans for patients. The new model can reduce the disease burden of patients and can contribute to the reasonable allocation of medical resources, which is significant for tertiary prevention.

Background: Doxorubicin (DOX) is a highly effective chemotherapeutic that is effective for various tumours. However, the clinical application of DOX has been limited by adverse reactions such as cardiotoxicity and heart failure. Since DOX-induced cardiotoxicity is irreversible, drugs to prevent DOX-induced cardiotoxicity are needed. Purpose: This study aimed to investigate the effect of total flavonoids of Selaginella tamariscina (P.Beauv.) Spring (TFST) on doxorubicin-induced cardiotoxicity. Methods: The present study established DOX-induced cardiotoxicity models in C57BL/6 mice treated with DOX (cumulative dose: 20 mg/kg body weight) and H9c2 cells incubated with DOX (1 mu M/l) to explore the intervention effect and potential mechanism of TFST. Echocardiography was performed to evaluate left ventricular functions. Heart tissue samples were collected for histological evaluation. Myocardial injury markers and oxidative stress markers were examined. Mitochondrial energy metabolism pathway associated proteins PPAR alpha/ PGC-1 alpha/Sirt3 were detected. We also explored the effects of TFST on endoplasmic reticulum (ER) stress and apoptosis. To further investigate the protective mechanism of TFST, we used the specific small interfering RNA MFN2 (siMFN2) to explore the effect of MFN2 on TFST against DOX-induced cardiotoxicity in vitro. Flow cytometry detected reactive oxygen species, mitochondrial membrane potential and apoptosis. Cell mitochondrial stress was measured by Seahorse XF analyser. Results: Both in vivo and in vitro studies verified that TFST observably alleviated DOX-induced mitochondrial dysfunction and ER stress. However, these effects were reversed after transfected siMFN2. Conclusion: Our results indicated that TFST ameliorates DOX-induced cardiotoxicity by alleviating mitochondrial dysfunction and ER stress by activating MFN2/PERK. MFN2/PERK pathway activation may be a novel mechanism to protect against DOX-induced cardiotoxicity.

Each Chinese medicine has its own properties and effects. However, the close connection between the medicinal properties and the effects of the medicine remains unclear. To export the scientific connection between the medicinal properties and efficacy of Rehmanniae Radix (RR), this study established a model and evaluated the therapeutic effects of RR on cold-heat syndrome to access the properties of RR, and then established a blood-heat syndrome model through the injection of rats with dry yeast combined with anhydrous ethanol. Related biochemical indicators (coagulation factors and central pyrogenic factor) were measured to assess the efficacy of RR. Finally, metabonomic technology was used to study the blood-cooling mechanism of RR from two aspects: medicinal properties and efficacy. The comprehensive results suggest that RR can significantly reduce the rectal temperature of blood-heat syndrome model rats and increase both the expression levels of coagulation factors (TNF-[Formula: see text], IL-1[Formula: see text], and IL-6) and the central pyrogenic factors (c-AMP, PGE-2). RR also cools the blood through regulating arginine, proline, phenylalanine, taurine, hypotaurine, sulfur, glycerophospholipid, primary bile acid metabolic pathways, and the tricarboxylic acid cycle. Therefore, RR plays the role of cooling blood by virtue of its cold property. The medicinal property of RR has a guiding effect on the clinical application. Moreover, the integrated metabolomic approach is a powerful tool for studying the properties and efficacy of Chinese medicine.

BACKGROUND:Rehmanniae Radix (RR), an herb with numerous pharmacological effects, is widely used in traditional Chinese medicine for the treatment of blood deficiency syndrome, either alone or in combination with other herbs. However, the mechanism by which processed Rehmanniae Radix (PRR) improves blood enrichment efficacy has not been clearly defined.METHODS:Ultra-performance liquid chromatography coupled to quadrupole time-of-flight mass (UPLC-Q-TOF/MS) and biochemical methods were combined to explore the hematopoietic functional mechanisms of PRR on blood deficiency in a rat model, as well as the potential active ingredient for blood enrichment efficacy. The pharmacological effects of PRR were evaluated on a rat blood deficiency model induced by cyclophosphamide in combination with 1-acetyl-2-phenylhydrazine. The blood routine index, including white blood cell (WBC), red blood cell (RBC), and platelet (PLT) counts, as well as hemoglobin (HGB) level, and the changing metabolite profile based on urine and serum were assessed. Nontargeted metabolomic studies, combined with biochemical analyses, were employed to clarify pharmacological mechanisms.RESULTS:PRR significantly increased the blood routine index levels and reversed the levels of SOD, GSH, and ATP. The PRR group was similar to the control group, as determined from the metabolic profile. All of the 60 biomarkers, representing the typical metabolic characteristics of the blood-deficient rat model, mainly involved energy metabolism dysfunction, the peripheral circulation system, and oxidative damage in the body. This improvement may be attributed to changes in polysaccharide and sixteen non-polysaccharide compounds in PRR, which were caused by processing RR with rice wine.CONCLUSIONS:The strategies of integrated metabolomic and biochemical analyses were combined, revealing the biological function and effective mechanism of PRR.

Objective(s): Gleditsiae spina (GS) is a natural antidepressant but its mechanisms of action remain unclear. In the present study, taxifolin (Tax) was selected to determine the role of flavonoids in the antidepressant effects of GS. Materials and Methods: Urine samples from C57BL/6 mice were analyzed based on ultra performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-Q/TOFMS). Then, we investigated the therapeutic effects of GS and Tax in depression models in vivo. An integrated metabolomic approach was used to examine the metabolic profiles of GS/Tax groups and corticosterone model groups (Cor). Metabolic networks in response to GS/Tax treatment were established for the comparison of antidepressant activities. Results: Corticosterone exposure significantly increased serum levels of corticosterone but decreased serum levels of 5-hydroxytryptamine and sucrose consumption (P<0.01). Treatment with GS and Tax improved all measured variables compared to those of the corticosterone-exposed group (P< 0.01). The antidepressant effects of GS and Tax involved the regulation of pentose and glucuronate interconversions, arginine and proline metabolism, phenylalanine metabolism, taurine and hypotaurine metabolism, and the citrate cycle. Conclusion: These findings indicate that flavonoids form the pharmacodynamic basis of the antidepressant effects of GS. Moreover, our findings highlight that integrated metabolomics provides a powerful tool to study the mechanisms and material basis of Chinese herbs.

Background: The traditional chinese medicine have medicinal property,and the property of the drug has been found to be mostly related to the body's substance and energy metabolism. Objectives: The present study intended to assess the drug property of Rehmanniae Radix Praeparata (PRR), called Shudihuang in traditional Chinese medicine and to discover its mechanism of cold syndrome in rats. Materials and Methods: Through the creation of a typical cold syndrome animal model, the appearance score, body weight, rectal temperatures, and survival rate of the animals treated were evaluated at different time points. Several indices in vivo correlated with glycolipid metabolism (glucose transporters (GLUT- 4), fructose phosphate kinase (PFK- 2), glycogen synthetase (GS), acetyl- CoA carboxylase 1 (ACC1), acetyl- CoA carboxylase 2 (ACC2), hydroxy methylglutaryl coenzyme A (HMG CoA) and hormone- sensitive lipase (HSL) were determined; Western blot is used to analyze of phosphorylated amp- dependent protein kinase (p- AMPK) appearance in liver tissues. Results: Compared with the normal group, the levels of GLUT- 4, PFK- 2, GS, ACC1, ACC2, HMG- CoA, and HSL in the model group all reduced significantly and p- AMPK protein expression diminished. Compared with the model group, PRR can significantly relapse to GLUT- 4, PFK- 2, GS, ACC1, ACC2, HMG- CoA, and HSL (P < 0.01 or P < 0.05) and rise the expression of p- AMPK. RR could suggestively diminution the level of these indices (P < 0.01 or P < 0.05). Conclusion: Drug property of PRR was inferred as trending to “heat and warm,” which still essential for further study. PRR may recover the metabolic function of the cold syndrome model rats by distressing the process of glycolipid metabolism which is by triggering the AMPK signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE:'Salt-processed Psoraleae Fructus & salt-processed Foeniculi Fructus' (sPF&sFF) is a common Chinese medicinal combination for treating diarrhoea. However, it is not clear how sPF and sFF work together, and why salt-processing is necessary.AIM OF THE STUDY:To investigate the compatibility mechanism of sPF&sFF and the influence of salt-processing on it.MATERIALS AND METHODS:Firstly, the metabolomics approach was appliedto screen the differential components between four (s)PF&(s)FF extracts, i.e., sPF&sFF, sPF&FF, PF&sFF, and PF&FF extracts. Then, an in vivo metabolomics study was carried out to filter critical metabolites reflecting the curative effects of (s)PF&(s)FF, and construct a metabolic network. Finally, a correlation analysis between chemical components in extracts and critical metabolites in vivo was performed to find out the synergistic and/or antagonistic effects between herbs as well as the influence of salt-processing.RESULTS:Salt-processing had a direct influence on the contents of chemical components in sPF and sFF extracts, and there existed positive/negative correlations between the content change of chemical components and the effects of critical metabolites. Therefore, salt-processing indirectly affected on these correlations and was (i) conducive to the positive effects of sPF and sFF on bile acids, making sFF play a synergistic role, thereby, sPF&sFF could perform better than sPF and other three combinations and effectively relieve the symptoms of fatty diarrhoea, osmotic diuresis, malnutrition, and weight loss; (ii) conducive to the positive effects of sPF on triacylglycerol, 12(S)-hydroxyeicosatetraenoic acid, cholesterol, and arachidonic acid, and adverse to that of sFF, making sFF play an antagonistic role, thereby, sPF&sFF could prevent a series of side effects caused by over-regulation and suitably relieve the symptoms of osmotic diuresis, polyuria, malnutrition, and weight loss; and (iii) adverse to the positive effects of sPF and sFF on thromboxane A2, sphinganine and sphingosine, making sFF play a synergistic role, thereby, sPF&sFF could prevent a series of side effects and moderately relieve the symptoms of metabolic diarrhoea and polyuria.CONCLUSIONS:Salt-processing indirectly affected on the correlations between chemical components in extracts and critical metabolites in vivo, and exhibited both conducive and adverse effects on the efficacy, making sPF and sFF cooperate with each other to moderately repair the metabolic disorders. Thereby, sPF&sFF could suitably relieve the diarrhoea and polyuria symptoms in the model and exert the most appropriate efficacy. Moreover, this novel strategy provided a feasible approach for further studying the compatibility mechanism of herbs.

A UPLC-Q-TOF/MS-based metabolomics study was carried out to explore the intervening mechanism of Corallodiscus flabellatus (Craib) B. L. Burtt (CF) extract on Alzheimer's disease (AD). The AD model group consisted of senescence-accelerated mouse prone 8 (SAMP8) mice, and the control group consisted of senescence-accelerated mouse resistant 1 (SAMR1) mice. UPLC-Q-TOF/MS detection, multivariate statistical analysis, and pathway enrichment were jointly performed to research the change in metabolite profiling in the urine of AD mice. The result suggested that the metabolite profiling of SAMP8 mice significantly changed at the sixth month compared with SAMR1 mice of the same age, and the principal component analysis (PCA) score scatter plots of the CF group closely resembled those of the control and positive drug (huperzine A, HA) group. A total of 28 metabolites were considered potential biomarkers associated with the metabolism of beta-alanine, glycine, serine, threonine, cysteine, methionine, arginine, proline, and purines in AD mice. Furthermore, the CF group was clustered with the control and positive group and was clearly separated from the model group in the heat map. In conclusion, significant anti-AD effects were firstly observed in mice after treatment with the CF extract, and the urinary metabolomics approach assisted with dissecting the underlying mechanism.

Chrysanthemum morifolium(Chr) is a traditional Chinese medicine (TCM) that has been used in the treatment of inflammation-linked diseases for hundreds of years. Naringenin (Nar) and apigenin (Api) are the major active components in aqueous extracts ofC. morifolium. The aim of our study was to clarify the roles of Chr, Nar and Api in ameliorating depression-like behaviour induced by corticosterone. First, the behavioural and biochemical indicators closely related to depression were examined to evaluate the therapeutic effects of Chr/Nar/Api on a depression model. Then, a metabolomics approach was utilized to screen for biomarkers and related pathways between a control group and Chr/Nar/Api groups. The comprehensive results revealed that Chr/Nar/Api exerted anti-depressant effects through interfering with tryptophan metabolism, arginine and prolinemetabolism, citrate cycle, niacin and niacinamide metabolism, phenylalanine metabolism, and alanine, aspartate and glutamate metabolism. The mechanism of Chr/Api/Nar in the treatment of depression was elucidated based on material and energy metabolism. Moreover, Nar could be used as a substitute for Chr for reversing depression-like behaviour, and Api was similar to a positive drug in terms of function on depression. The integrated metabolomics approach demonstrated here should be an effective method for interpreting the function of herbs from TCM and clarifying the mechanism of their components in future studies.