Exploration of the intestinal flora to reveal the important contribution of Radix Astragali to Huangqi Jianzhong Tang in treating chronic atrophic gastritis rats.

Radix Astragali (Huangqi in Chinese, HQ) is a commonly used Chinese herbal medicine for thousands of years. In this study, A classic prescription Huangqi Jianzhong tang (HQJZ) was selected to evaluate the important effect of HQ on rats with chronic atrophic gastritis (CAG) from the perspective of intestinal flora in cecal contents samples. Traditional pharmacological indicators, including weight change, pathological examination and biochemical indicators showed that HQ exerted favorable contribution to HQJZ against CAG, where the efficiencies of HQ and HQJZ were better than HY (HQJZ prepared without HQ). An accurate strategy was adopted to screen out the differential metabolites in the metabolomis analysis of intestinal flora in cecal contents samples based on the optimal screening factors, including VIP (importance of variables in projection), FC (fold change), AUROC (area under the receiver operating characteristic curve) and -ln(p-value), which were evaluated based on their interpreting, grouping, and predicting abilities of the performed orthogonal partial least-squares-discriminate analysis (OPLS-DA) models. Ten altered differential metabolites were obtained and associated with the intestinal flora, which HQ exerted the important metabolic contributions to HQJZ. The efficacy on the diversity of intestinal flora and their correlations with the altered metabolites further showed the important role of HQ in HQJZ composition. This work provided valuable approach for looking for potential biomarkers associated with metabolomics research with more accuracy, and provided new insights into the mechanisms to explain the efficacy of HQ contributing to HQJZ formula.

Integration of diagnostic ions, molecular network and chemometrics to illustrate the chemical mechanism of Radix Astragali processed with honey.

Radix Astragali (RA) is one of the most frequently used traditional Chinese medicine (TCM) in China, and honey-processed RA (HRA) is its common processing product. Thus far, their comprehensive chemical differences are not well understood. In this work, an integrated approach using Ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) combined with diagnostic ions, molecular network (MN) and chemometrics was established to profile their chemical characterizations and illustrate the chemical mechanism of RA processed with honey. A total of 226 compounds were tentatively identified including 50 flavonoid glycosides, 26 flavonoid aglycone, 56 saponins, 30 organic acids, 18 amino acids, 3 coumarins and 43 other compounds, of which 33 compounds were characterized according to MN. Their chemical differences were further investigated by integrating of multivariate statistical analysis, student's t-test analysis, linear regression analysis and MN. Consequently, multivariate statistical analysis showed that the raw and processed RA were different form each other. Besides, 33 different compounds were found to be significantly altered by student's t-test analysis. Apart from this, linear regression analysis indicated 42 and 120 compounds underwent the significant varieties. The potential chemical reactions induced by honey-processing, such as possible hydrolysis reactions and isomerization reactions, were speculated based on these variations coupled the areas changes of the nodes in MN. This study provided an efficient strategy to illustrate the chemical mechanism of TCM processing.

Integrating metabolomics, 16S rRNA sequencing, network pharmacology, and metorigin to explore the mechanism of Cinnamomi Cortex in treating chronic atrophic gastritis rats.

BACKGROUND: Cinnamomi cortex called as Rougui (RG) in Chinese was a widely used food-medicine homology. RG has the potential to treat chronic atrophic gastritis (CAG), a disease with widespread impact in the Chinese population. PURPOSE: This study aimed to explore its mechanism against CAG based on amalgamated strategies. METHODS: Network pharmacology was used to predict the potential effective components and the core targets of RG against CAG based on the comprehensive chemical characterization using UHPLC-Q/TOF MS (ultra high performance liquid chromatogramphy-quadrupole/time-of-flight mass spectrometry). The CAG animals model were further used to validate its pharmacodynamics, of which gut microbiota of caecal contents were analyzed by integrating metabolomics, 16S rRNA sequencing, Metorigin metabolite traceability analysis and molecular docking to explore its action mechanism. RESULTS: Network pharmacology firstly predicted the efficacy of RG was attributed to four effective components and seven targets. Metabolomics of caecal contents in CAG rats revealed primary bile acid biosynthesis was its targeted metabolic pathway associated with the metabolism of gut microbiota coupled with Metorigin traceability analysis. 16S rRNA sequencing showed that RG treated CAG by regulating the imbalance of gut microbiota. Molecular docking further confirmed that the effective components of RG could intervene with potential targets, metorigin analysis pathway, and key enzymes of gut microbiota metabolic pathways. CONCLUSION: Our results proved that RG exerted favorable effect on CAG. The four active ingredients (quercetin, kaempferol, oleic acid, and (-)-epicatechin) of RG were the key to exert drug effect, which could targeted the core target of CAG, primary bile acid biosynthesis and intestinal flora metabolic pathways.

Amalgamated Pharmacoinformatics Study to Investigate the Mechanism of Xiao Jianzhong Tang against Chronic Atrophic Gastritis.

BACKGROUND: Traditional Chinese medicine (TCM) Xiaojianzhong Tang (XJZ) has a favorable efficacy in the treatment of chronic atrophic gastritis (CAG). However, its pharmacological mechanism has not been fully explained. OBJECTIVE: The purpose of this study was to find the potential mechanism of XJZ in the treatment of CAG using pharmacocoinformatics approaches. METHODS: Network pharmacology was used to screen out the key compounds and key targets, MODELLER and GNNRefine were used to repair and refine proteins, Autodock vina was employed to perform molecular docking, ΔLin_F9XGB was used to score the docking results, and Gromacs was used to perform molecular dynamics simulations (MD). RESULTS: Kaempferol, licochalcone A, and naringenin, were obtained as key compounds, while AKT1, MAPK1, MAPK14, RELA, STAT1, and STAT3 were acquired as key targets. Among docking results, 12 complexes scored greater than five. They were run for 50ns MD. The free binding energy of AKT1-licochalcone A and MAPK1-licochalcone A was less than -15 kcal/mol and AKT1-naringenin and STAT3-licochalcone A was less than -9 kcal/mol. These complexes were crucial in XJZ treating CAG. CONCLUSION: Our findings suggest that licochalcone A could act on AKT1, MAPK1, and STAT3, and naringenin could act on AKT1 to play the potential therapeutic effect on CAG. The work also provides a powerful approach to interpreting the complex mechanism of TCM through the amalgamation of network pharmacology, deep learning-based protein refinement, molecular docking, machine learning-based binding affinity estimation, MD simulations, and MM-PBSA-based estimation of binding free energy.

Comparison of two different integrated method of pharmacokinetics by the integrated pharmacokinetic research of fangji huangqi decoction.

Traditional Chinese medicine (TCM) is characterized by its multiple components. The utilization of mathematical statistical methods to integrate the pharmacokinetics of monomer components can provide a comprehensive understanding of the holistic pharmacokinetic process of TCM. Two distinct integrated methods, namely the correlation coefficient method and the AUC-based weight coefficient method, were employed in this study to elucidate and compare their differences in the integrated pharmacokinetic research of Fangji Huangqi decoction (FHD). FHD is commonly used in clinical practice to treat the nephrotic syndrome. Firstly, one-dose FHD was given to doxorubicin-induced nephropathy (DN) rats, and the prototype compounds of FHD and their metabolites in plasma were qualitatively and semi-quantitatively analyzed by UHPLC-MS/MS. Secondly, the efficacy of FHD was quantitatively characterized by the relative distance method based on metabolomics. The correlation coefficients were obtained by analyzing the correlation between efficacy (relative distance values) and the content of compound, and they were subsequently used for the model integration (correlation coefficient method). Thirdly, the effective compounds of FHD treating DN were screened by integrating network pharmacology and molecular docking, and they were used for another integrated pharmacokinetic model by AUD-based weight coefficient method. Finally, the 2 integrated methods and the 2 integrated pharmacokinetic models were compared. In this study, 30 prototype compounds and 41 metabolites of FHD in plasma were identified, and the pharmacokinetic curve of 18 prototype compounds were built. The efficacy of FHD in the treatment of DN has been relatively quantitation. The 2 established integrated pharmacokinetic models of FHD indicated that the correlation coefficient method was the optimal approach for conducting the integrated pharmacokinetic research on the TCM with unknown effective compounds, whereas the AUC-based coefficient method was suitable for the TCM with the clear effective compounds. The integrated pharmacokinetic models indicated that FHD had high bioavailability and an absorption peak at about 6 h after administration, indicating that the 6 h after administration was the critical period of FHD treating DN. This research would be helpful for the pharmacological and pharmacokinetic research of FHD, and provide a method reference for the integrated pharmacokinetic research of TCM.

Rapid chemical characterization of Danggui Buxue decoction using processed Astragali Radix and Angelicae Sinensis Radix based on diagnostic ion and molecular network.

Danggui Buxue decoction (DBD), a classic prescription of traditional Chinese medicine (TCM) for invigorating qi and generating blood, contains honey-processed Astragali Radix (HAR) and wine-processed Angelicae Sinensis Radix (WDG) in its original prescription. In this study, the compositions of DBD, WDG, and HAR were characterized using ultra-high-performance liquid chromatography coupled with the quadrupole-time-of-flight tandem mass spectrometry technique in combination with molecular network and diagnostic ion strategies. Finally, 200 compounds were identified in DBD, 114 compounds were identified in WDG, and 180 compounds were identified in HAR; there were 48 common compounds in total. The results demonstrated that compatibility led to changes in the chemical composition of TCM, and the qualitative method used in this study provided an effective data processing strategy for the characterization of components and the database for the study of the compounding mechanism of TCM.

Bile acid metabolism involved into the therapeutic action of Xiaojianzhong Tang via gut microbiota to treat chronic atrophic gastritis in rats.

BACKGROUND: As a classical traditional Chinese medicine (TCM), Xiaojianzhong Tang (XJZ) is effective in treating chronic atrophic gastritis (CAG). However, the pharmacological mechanism of XJZ has not been fully explained. PURPOSE: The purpose of this study was to investigate the mechanism of XJZ against CAG rats via gut microbiome using a multi-omics approach. METHODS: The rat cecal contents were analyzed through the integration of an untargeted metabolomic approach based on ultra-high performance liquid chromatography coupled with the quadrupole-time of flight mass spectrometry (UHPLC-QTOF-MS) and 16S rRNA gene sequencing. Finally, the interaction of differential metabolites with bile acid (BA)-related targets was verified by molecular docking. RESULTS: A new strategy was adopted to screen out the differential metabolites based on the comprehensive evaluation of VIP, |log2(FC)|, -ln(p-value) and ǀp(corr)ǀ. As results, XJZ showed favor regulations on the screened metabolites, cholic acid, deoxycholic acid, glycoursodeoxycholic acid, taurochenodesoxycholic acid, docosahexaenoic acid and L-isoleucine. The 16S rRNA gene sequencing analysis showed that XJZ could regulate gut microbiota disturbances in CAG rats, especially bile acid (BA) metabolism-related bacteria (Butyricimonas, Desulfovibrio, Bacteroides, Parabacteroides, Acetobacter and Alistipes). Molecular docking further showed that the differential metabolites regulated by XJZ had a good docking effect on BA-related targets. CONCLUSION: The current work indicated that XJZ's therapeutic action was strongly linked to BA-related microorganisms and metabolic processes. These findings provided new insights into the effects of XJZ for the treatment of CAG.

Exploration of the main effective constituent and the mechanism in Astragali Radix in the treatment for doxorubicin-induced nephropathy by integrating metabolomics and molecular docking.

ETHNOPHARMACOLOGICAL RELEVANCE: Astragali Radix (AR) is the dried root of Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao or A. membranaceus (Fisch.) Bge. AR was the main medicine in a Chinese traditional prescription called Fangji Huangqi Decoction, and it has been used to treating nephrotic syndrome (NS) for thousands of years in China. In recent years, AR has been evidenced to have anti-inflammatory activity, antihyperglycemic activity, antioxidant activity, etc. There are two mainstream commodities for ARs in the market including the imitation wild AR and transplanted AR. However, it is not clear whether the imitation wild AR or transplanted AR and which kind of component, astragalus saponin, astragalus flavonoid or astragalus polysaccharide, makes a bigger contribution in treating NS. And the exact molecular mechanism is not fully understood. AIM OF THE STUDY: To explore which kind of AR and which kind of component in AR makes the bigger contribution in treating NS, and exploring the molecular mechanism. MATERIALS AND METHODS: Firstly, HPLC-UV/ELSD was used for quantitative determination of the constituents in different ARs. Secondly, the efficacy of different ARs treating doxorubicin-induced nephropathy (DN) was compared by metabolomics. Thirdly, the protective effects of different constituents from ARs on the damage of MPC5 cells induced by adriamycin are validated. Finally, the effective constituents and mechanism of ARs against doxorubicin-induced nephropathy were investigated by network pharmacology and molecular docking. RESULTS: Quantitative determination experiment and pharmacological experiment indicated that the AR produced from Gansu province (China) (transplanted AR) with a higher proportion of total saponins, has better efficacy in the treatment for DN. And the cell experiment validated the result that astragalus saponins has the better efficacy in protecting the podocyte against injury than astragalus flavonoids and polysaccharides. The network pharmacology and molecular docking study indicated that astragalus saponins were the main constituent of AR in the treatment for DN. The mechanism may involve in GnRH signaling pathway, VEGF signaling pathway and metabolic pathways, especially of bilirubin metabolism. CONCLUSIONS: Transplanted AR has better efficacy in the treatment for NS than imitation wild AR, astragalus saponins have better efficacy in the treatment for NS than astragalus flavonoids and polysaccharides.

Metabolite Identification of Huangqi Jianzhong Tang in Rat Urine and Feces after Oral Administration Based on UHPLC-Q-Exactive-MS.

Huangqi Jianzhong Tang (HQJZ), a famous traditional Chinese medicine formula, is widely used in the treatment of gastrointestinal diseases in China. In this study, an ultra-high-performance liquid chromatography coupled with quadrupole-Exactive mass spectrometry was used to identify the metabolites in rat urine and feces after oral administration of HQJZ coupled with Compound Discover 3.0 software. The possible metabolic pathways were calculated and deduced based on 71 previous detected prototype compounds. As results, 88 compounds were identified in urine and feces, respectively. In urine sample, we identified 20 prototype compounds and 68 related metabolites. Meanwhile, a total of 21 prototype compounds and 67 related metabolites were identified in feces. Among them, flavonoids and saponins were the main ingredients in vivo. Further, we also speculated that HQJZ experienced oxidation, reduction, acetylation, methylation and glucuronic acid reaction in urine and feces. This study established a reliable method for the detection of prototype components and metabolites of traditional Chinese medicines, which would provide helpful information for further research into the active substances of HQJZ.

Rapid Quantitative Analysis of 19 Bioactive Components in Fangji Huangqi Decoction Based on UHPLC-MS/MS.

Fangji Huangqi Decoction (FHD) is a classic prescription of traditional Chinese medicine which is recorded in "Jin Gui Yao Lue". The purpose of this study is to develop a method for simultaneous determination multicomponent in FHD. The separation of the 19 compounds that included calycosin, calycosin-7-O-ß-D-glucoside, formononetin, ononin, methylnissolin, methylnissolin-3-O-glucoside, isomucronulatol, tetrandrine, fangchinoline, atractylenolide-I, atractylenolide-III, liquiritigenin, liquiritin, isomucronulatol-7-O-ß-D-glucoside, astragaloside-I, astragaloside-II, astragaloside-III, astragaloside-IV and glycyrrhetinic acid were achieved by linear gradient elution. The 19 components were identified by comparing the chromatographic peaks with the reference compounds and were quantitatively analyzed by multiple reaction monitoring. This method was strict validated with recovery (96.10-101.70%), precision [relative standard deviation (RSD), 1.34-3.34%], stability (RSD, 1.49-3.80%) and repeatability (RSD, 1.60-3.49%), respectively. All the compounds showed good linearities (R2 > 0.999). The limit of detection (LOD) and limit of quantitation (LOQ) for the 19 compounds were in the range of 0.03-0.27 µg/mL (LODs) and 0.05-1.23 µg/mL (LOQs). The correlation analysis indicated that astragalus flavonoids were negatively correlated with astragalosides, tetrandrine and their corresponding flavonoid glycosides, and atractylenolides were positively correlated with astragalosides and fangchinoline. This method proved to be reliable and effective, which would give a helpful basis for the quality control, pharmacological and pharmacokinetic of FHD.

Integrated pharmacokinetics of Huangqi Jianzhong Tang in normal and chronic atrophic gastritis rats.

Huangqi Jianzhong Tang (HQJZ) is a famous traditional Chinese medicine formula widely used in the treatment of gastrointestinal diseases in China. In this study, an ultra-performance liquid chromatography-mass spectrometry (UHPLC-MS/MS) was used to study the pharmacokinetics of 12 prototypical components and one metabolite in HQJZ in normal and chronic atrophic gastritis rats. The results showed that the area under the concentration-time curve and peak concentration of most flavonoids and flavonoid glycosides were decreased, and the half-life and mean residence time were significantly increased, which indicated that the absorption of drugs in disease was decreased less and for longer in vivo. Then, an integrated pharmacokinetic study was carried out using the pharmacokinetic parameter model integration of each component. The results showed that the absorption of drugs in vivo with disease was reduced, and the absorption speed of flavonoids and flavonoid glycosides was accelerated. This study will provide the basis for the clinical medication safety of HQJZ.

Regulation of gut microbiota of Astragali Radix in treating for chronic atrophic gastritis rats based on metabolomics coupled with 16S rRNA gene sequencing.

Astragali Radix (HQ), a common traditional Chinese medicine (TCM), is widely used to treat chronic atrophic gastritis (CAG). However, its mechanism in treating CAG is still not clear. Accumulating evidence highlights the link between gut microbiota and CAG. We hypothesized that the gut microbiota might be involved in the effect of HQ. Ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC-Q-TOF/MS) based metabolomics and 16S rRNA gene sequencing techniques of the cecal contents were applied to study its mechanisms. As a result, nine metabolites and fifteen gut microbiotas changed significantly in cecal contents samples between control group and model group. Among them, two metabolites (7-keto-3A ·12-α-hydroxyalkanoic acid and deoxycholic acid) and two gut microbiota genera (Acetobacter and Escherichia), had the most obvious callback effect after the administration of HQ. Sixty-seven correlated pairs exhibited the significant link between the involved metabolites and gut microbiotas through the correlation analysis, where two strong correlation pairs: Tetrahydrohydroxone âˆ¼ Bacteroides (r = 0.895) and Deoxycholic acid âˆ¼ Acetobacter (r = -0.843) were regulated by HQ. The results showed that HQ had the potential protection from metabolic perturbation involved into gut microbiotas induced by CAG. Two gut microbiotas, Acetobacter and Escherichia, and two metabolites, 7-keto-3A ·12-α-hydroxyalkanoic acid and deoxycholic acid were the potential targets of HQ.

Studies on the compatibility mechanism and material basis of Danggui Buxue Decoction against anemia mice using metabonomics and network pharmacology.

OBJECTIVES: To reveal the compatibility mechanism and material basis of Danggui Buxue decoction (DBD) against anaemia. METHODS: UHPLC-Q-Exactive-MS based serum metabonomics was applied to decipher the compatibility of DBD against anaemia mice. Meanwhile, network pharmacology was used to reveal the material basis of DBD based on the obtained differential metabolites. KEY FINDINGS: Metabonomic results indicated that 17 serum differential metabolites were closely related to anaemia. DBD, Huangqi (HQ) and Danggui (DG) could significantly ameliorate 13, 6 and 4 serum metabolites in anaemia mice, respectively. 17 serum differential metabolites were linked 140 corresponding targeted genes obtained by Metscape. In addition, 6649 targets genes related anaemia were obtained by network pharmacology. At last, six important targets genes were screened as hopeful targets for the treatment of anaemia through integrating them. Molecular docking further illustrated that eight active components of DBD including mairin, hederagenin, etc. played important roles in treating anaemia. CONCLUSIONS: DBD produced the best effect by compatibility with HQ and DG in treating anaemia. The approach provided the insights into the compatibility mechanism and material basis of TCM in treating anaemia coupling network pharmacology.

Mitochondria metabonomics of Huangqi Jianzhong Tang against chronic atrophic gastritis.

Huangqi Jianzhong Tang (HQJZ) is a representative prescription used for clinical treatment of chronic atrophic gastritis (CAG) in Chinese medicine. Our previous study had revealed that energy regulation was one of the important mechanisms of HQJZ action against CAG. In this study, ultra-high-performance liquid chromatography coupled with quadrupole-Exactive mass spectrometry (UHPLC-Q-Exactive MS) based metabonomics was used to find the potential mitochondrial biomarkers and metabolic pathways of HQJZ in CAG rats, which focused on a specific organelle (mitochondria) isolated from gastric tissue samples. A total of 16 biomarkers from CAG tissues were identified with 11 of these significantly regulated by HQJZ treatment. These biomarkers was mainly involved in glycine, serine, and threonine metabolism; aminoacyl-tRNA biosynthesis metabolism; and taurine and hypotaurine metabolism. Our results show that HQJZ could protect from CAG by altering the mitochondrial function. These findings deepen our understanding of the mitochondrial metabolic changes that occur with CAG and shine a light on the mechanism of HQJZ.

Uncovering the mechanism of Astragali Radix against nephrotic syndrome by intergrating lipidomics and network pharmacology.

BACKGROUND: Astragali Radix (AR), a common Traditional Chinese Medicine (TCM), is commonly used for treating nephrotic syndrome (NS) in China. At present, the research on the efficacy of AR against NS is relative clearly, but there are fewer researches on the mechanism. PURPOSE: The aim of this study was to evaluate the potential beneficial effects of AR in an adriamycin-induced nephropathy rat model, as well as investigate the possible mechanisms of action and potential lipid biomarkers. METHODS: In this work, a rat model of NS was established by two injections of ADR (3.5 + 1 mg/kg) into the tail vein. The potential metabolites and targets involved in the anti-NS effects of AR were predicted by lipidomics coupled with the network pharmacology approach, and the crucial metabolite and protein were further validated by western blotting and ELISA. RESULTS: The results showed that 22 metabolites such as l-carnitine, LysoPC (20:3), and SM (d18:1/16:0) were associated with renal injury. Moreover, SMPD1, CPT1A and LCAT were predicted as lipids linked targets of AR against NS, whilst glycerophospholipid, sphingolipid and fatty acids metabolism were involved as key pathways of AR against NS. Besides, AR could play a critical role in NS by improving oxidative stress, inhibiting apoptosis and reducing inflammation. Interestingly, our results indicated that key metabolite l-carnitine and target CPT1 were one of the important metabolites and targets for AR to exert anti-NS effects. CONCLUSION: In summary, this study offered a new understanding of the protection mechanism of AR against NS by network pharmacology and lipidomic method.

Potential quality evaluation approach for the absolute growth years' wild and transplanted Astragali Radix based on anti-heart failure efficacy.

The quality of Astragali Radix (AR) was closely related to the growth period. However, the current commodity grades of AR were only divided by diameter but not directly related to the growth period, which leads to the contradiction between the grade standard and the quality evaluation index. Therefore, solving this problem will be the key for the quality evaluation of AR. The present study established a potential quality evaluation approach for the absolute growth years' wild Astragali Radix (WAR) and transplanted Astragali Radix (TAR) based on the chemical components and anti-heart failure efficacy through adopting a bare-handed sections approach to rapidly identify the growth years of WAR. In this study, the absolute growth years of WAR were obtained by identifying the growth rings of 1-6 growth years root through the methods. The contents of flavonoids and saponins in 2-6 growth years' WAR were determined by HPLC-UV-ELSD. The contents of 12 chemical components and the anti-fatigue failure effects of WAR (4-year-old) and TAR were compared on rat models of heart failure induced by doxorubicin. Meanwhile, NMR-based untargeted metabolomics studies were performed to investigate the regulative effects of WAR and TAR. The result shows that the numbers of growth rings were consistent with the actual growth periods of AR. The HPLC-UV-ELSD determination indicated that the content of total flavonoids in WAR was significantly higher than that in TAR. Pharmacodynamics analysis revealed that the effects of WAR on cardiac function parameters (EF, FS and LVIDs), contents of serum CK and BNP were superior to those of TAR. 13 metabolites of heart were identified that had a higher rate of change in WAR group than TAR. Overall, a rapid identification method for the growth years of WAR was established, and the fact that WAR were significantly better than TAR in the heart failure rats was first proved in the paper. This study provided a scientific basis for establishing a novel commodity specification and grade of AR for clinical rational drug use.

An integrated transcriptomics and network pharmacology approach to exploring the mechanism of adriamycin-induced kidney injury.

BACKGROUND AND AIMS: Adriamycin nephropathy model (AN), a rodent model of nephrotic syndrome disease that was caused by the nephrotoxicity of adriamycin, has been widely used for pharmacodynamic evaluation of traditional Chinese medicine (TCM) in the treatment of kidney injury. Although some studies have clearly shown the pathological process of AN, the mechanism of kidney injury have not been systematically investigated. METHODS: The reliability of AN was evaluated by weight, urinary protein quantitation, serum biochemical and histopathological examination. Transcriptomic sequencing combined with network pharmacology were used to elucidate the molecular mechanism of AN, and cell experiment combined with real-time quantitative PCR (RT-qPCR) and was used to validate the accuracy of transcriptomic sequencing result and KEGG pathways. RESULTS: Network analysis result showed that Mapk10 and Ptgs2 played important roles in the development of adriamycin-induced kidney injury. KEGG pathway analysis showed that the mechanism of kidney injury may be related to the regulation of biosynthesis of unsaturated fatty acids, complement and coagulation cascades, PPAR signaling pathway and PI3K-AKT signaling pathway. CONCLUSION: These results provide a new insight into the deep research on the mechanism of kidney injury, and provide an experimental basis for finding drug targets for the treatment of AN.

Network pharmacology research of Astragali Radix in treating chronic atrophic gastritis rats based on mitochondrial metabonomics.

Astragali Radix (HuangQi), one important traditional Chinese herb, is used for treatment of chronic atrophic gastritis (CAG). To comprehensively evaluate its regulation on CAG, a mitochondria-specific metabonomics was applied to reveal its action on energy metabolism based on ultra high performance liquid chromatography coupled with quadrupole - Exactive mass spectrometry. 16 related metabolites from mitochondria samples were served as potential biomarkers of CAG. Nine out of them were significantly regulated by HuangQi. Combining with network pharmacology, three active components from HuangQi and 3 mitochondrial metabolites exerted better docking abilities with 56 predicted targeted proteins based on SystemsDock, which were involved into multiple biological abnormities including mitochondrion dysfunction. The results demonstrated that mitochondrial energy metabolism played crucial role contributing to HuangQi against CAG, which was one important mechanism of HuangQi.

Urinary metabolomics research for Huangqi Jianzhong Tang against chronic atrophic gastritis rats based on 1 H NMR and UPLC-Q/TOF MS.

OBJECTIVES: As a traditional Chinese medicine (TCM), Huangqi Jianzhong Tang (HQJZ) has a good efficacy in treating chronic atrophic gastritis (CAG). Our objective was to determine its mechanism based on the urine comprehensive metabolome. METHODS: In the study, a metabolomic approach was applied to reveal the efficacy of HQJZ on the constructed CAG rats coupled with proton nuclear magnetic resonance (1 H NMR) and ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF MS). KEY FINDINGS: The results showed the regulatory effect of HQJZ on urinary metabolism disorder in CAG rats was similar to the positive drug teprenone. Nineteen and 16 potential biomarkers related to CAG were detected by NMR and UPLC-Q/TOF MS, respectively. Thirty-two urine metabolites were significantly regulated by HQJZ treatment. Combined with MetPA and partial least square regression analysis (PLS-RA), three metabolic pathways of valine, leucine and isoleucine, TCA cycle, and glycine, serine and threonine metabolism were the most relevant pathways for HQJZ treatment. CONCLUSIONS: The main mechanism of HQJZ might be due to the balance of energy consumption, inflammatory inhibition, improvement of the immune system and oxidative stress on the constructed CAG rats. These findings provided comprehensive metabolic information of TCM by parallel measurements by LC-MS and NMR.

UHPLC Q-Exactive MS-based spleen metabolomics and lipidomics to explore the effect mechanisms of Danggui Buxue Decoction in anemia mice.

Danggui Buxue Decoction (DBD), a famous traditional Chinese medicine (TCM), is often used to treat anemia in China. However, its underlying therapeutic mechanism is unclear. Through the analysis of body weight, spleen and thymus indexes, peripheral blood routine and pathological section of femur, it was obviously that DBD could significantly improve acetylphenylhydrazine (APH) + cyclophosphamide (CTX) induced anemia mice in the present work. Ultra high performance liquid chromatography coupled with quadrupole - Exactive mass spectrometry (UHPLC Q-Exactive MS) based metabolomics and lipidomics was further utilized to screen out differential spleen metabolites associated with DBD treatment. A total of 26 differential metabolites including 8 polar metabolites and 18 lipids were firstly obtained to relate with anemia mice. 7 polar metabolites and 10 lipids among them were reversed by DBD, which the regulation of pyrimidine metabolism and glycerophospholipid metabolism were mainly associated to the anti-anemia effect of DBD based on MetaboAnalyst analysis. Through random forest analysis (RF), ROC analysis and pearson matrix correlation, three metabolites, cytosine, uracil and PC (o-16:1(9Z)/20:0), were further screened out as the potential pharmacodynamic biomarkers associated with the efficacy of DBD. This study provided a methodological reference for the study of the mechanism of TCM.