Target Screen of Anti-Hyperuricemia Compounds from Cortex Fraxini In Vivo Based on ABCG2 and Bioaffinity Ultrafiltration Mass Spectrometry.

The ATP-binding cassette (ABC) transporter ABCG2 is a significant urate transporter with a high capacity, and it plays a crucial role in the development of hyperuricemia and gout. Therefore, it has the potential to be targeted for therapeutic interventions. Cortex Fraxini, a traditional Chinese medicine (TCM), has been found to possess anti-hyperuricemia properties. However, the specific constituents of Cortex Fraxini responsible for this effect are still unknown, particularly the compound that is responsible for reducing uric acid levels in vivo. In this study, we propose a target screening protocol utilizing bio-affinity ultrafiltration mass spectrometry (BA-UF-MS) to expediently ascertain ABCG2 ligands from the plasma of rats administered with Cortex Fraxini. Our screening protocol successfully identified fraxin as a potential ligand that interacts with ABCG2 when it functions as the target protein. Subsequent investigations substantiated fraxin as an activated ligand of ABCG2. These findings imply that fraxin exhibits promise as a drug candidate for the treatment of hyperuricemia. Furthermore, the utilization of BA-UF-MS demonstrates its efficacy as a valuable methodology for identifying hit compounds that exhibit binding affinity towards ABCG2 within TCMs.

Stromal cells-specific retinoic acid determines parturition timing at single-cell and spatial-temporal resolution.

The underlying mechanisms governing parturition remain largely elusive due to limited knowledge of parturition preparation and initiation. Accumulated evidences indicate that maternal decidua plays a critical role in parturition initiation. To comprehensively decrypt the cell heterogeneity in decidua approaching parturition, we investigate the roles of various cell types in mouse decidua process and reveal previously unappreciated insights in parturition initiation utilizing single-cell RNA sequencing (scRNA-seq). We enumerate the cell types in decidua and identity five different stromal cells populations and one decidualized stromal cells. Furthermore, our study unravels that stromal cells prepare for parturition by regulating local retinol acid (RA) synthesis. RA supplement decreases expression of extracellular matrix-related genes in vitro and accelerates the timing of parturition in vivo. Collectively, the discovery of contribution of stromal cells in parturition expands current knowledge about parturition and opens up avenues for the intervention of preterm birth (PTB).

Identification of desoxyrhapontigenin as a novel antiviral agent against congenital Zika virus infection.

Zika virus (ZIKV) infection arises as a global health threat owing to its association with Guillain-Barre syndrome and microcephaly in adults and fetuses since the most recent epidemics. Although extraordinary efforts have been underway globally to identify safe and effective treatments for ZIKV, therapeutic progressions seem to remain stagnant, especially for treating congenital ZIKV infection. Bio-compounds from medicinal plants evolutionarily optimized as drug-like molecules offer eligible sources of pharmaceuticals and lead drugs to fight against viral infections. Here, we identified desoxyrhapontigenin (DES), a naturally occurring bioactive product, as the strongest inhibitory compound against ZIKV infection among six conventional polyphenols in vitro. We also leveraged the trophoblast cell line, human trophoblast stem cells, and complex placental organoid models to provide solid evidence to support the anti-ZIKV bioactivity of DES. Notably, DES treatment effectively reduced the ZIKV burden in serum and target tissues, and correspondingly improved ZIKV-induced pathologic changes including weight loss, tissue inflammation, cell apoptosis, and adverse pregnancy outcomes, while it did not lead to obvious toxicity in both adult and pregnant mice. Furthermore, mechanistic studies revealed that DES could suppress ZIKV entry via dual mechanisms of direct targeting ZIKV E proteins and downregulating putative ZIKV receptors. These findings elucidate a previously unappreciated protective role of desoxyrhapontigenin against ZIKV infection both in vitro and in vivo, which shed light on the development of a novel and potent treatment for congenital ZIKV infection.

Detection and Removal of Aristolochic Acid in Natural Plants, Pharmaceuticals, and Environmental and Biological Samples: A Review.

Aristolochic acids (AAs) are a toxic substance present in certain natural plants. Direct human exposure to these plants containing AAs leads to a severe and irreversible condition known as aristolochic acid nephropathy (AAN). Additionally, AAs accumulation in the food chain through environmental mediators can trigger Balkan endemic nephropathy (BEN), an environmental variant of AAN. This paper presents a concise overview of the oncogenic pathways associated with AAs and explores the various routes of environmental exposure to AAs. The detection and removal of AAs in natural plants, drugs, and environmental and biological samples were classified and summarized, and the advantages and disadvantages of the various methods were analyzed. It is hoped that this review can provide effective insights into the detection and removal of AAs in the future.

Value of Chuanjin Qinggan decoction in improving the depressive state of patients with herpes zoster combined with depression.

BACKGROUND: Western medicine is beneficial for the recovery of neurological function in patients with depression, but some patients experience side effects such as headaches, dizziness, nausea, gastrointestinal symptoms, insomnia, and cardiac dysfunction. In recent years, integrative medicine has achieved positive results in the treatment of various diseases. AIM: To study Chuanjin Qinggan decoction combined with selective serotonin reuptake inhibitors (SSRIs) in patients with herpes zoster complicated by depression. METHODS: Patients with herpes zoster complicated by depression who were treated at the Yantai Hospital of Traditional Chinese Medicine from January 2021 to December 2022 were retrospectively selected as research participants. Among them, 43 patients with herpes zoster complicated by depression who received SSRI treatment between January and December 2021 were assigned to the Western medicine group, while those who received combined treatment of traditional Chinese and Western medicine between January and December 2022 were assigned to the combined group. Both groups were treated for eight weeks. The degree of pain, effect of herpes zoster treatment, degree of improvement in depressive symptoms, serum neurotransmitter levels, sleep quality, and occurrence of adverse reactions were compared between the two groups. RESULTS: We found that after eight weeks of drug treatment, the two treatment schemes achieved differing efficacy. In further comparison, we found that, compared with patients treated with SSRIs alone, patients treated with Chuanjin Qinggan decoction combined with SSRIs showed more significant improvement in depression and a greater increase in dopamine and 5-hydroxytryptamine levels after treatment (P < 0.05). Patients treated with Chuanjin Qinggan decoction combined with SSRIs also experienced lower pain, better treatment efficacy for herpes zoster, better sleep quality, and a lower incidence of adverse reactions compared to those treated with SSRIs alone (P < 0.05). All minor adverse reactions occurring during treatment were resolved after symptomatic treatment. CONCLUSION: The treatment scheme of Chuanjin Qinggan decoction combined with SSRIs can improve the psychological state of patients with herpes zoster complicated by depression and alleviate adverse reactions.

Efficacy of Fangfeng Tongsheng Granule Combined with Levocetirizine in the Treatment of Chronic Urticaria and Its Effect on Serum Complement, IL-4, IgE, and IFN-γ Levels in Patients.

Objectives: To investigate the efficacy of Fangfeng Tongsheng granule combined with levocetirizine in the treatment of chronic urticaria and its effect on serum complement, interleukin (IL)-4, immunoglobulin E (IgE), and interferon-γ (IFN-γ) levels in patients. Methods: A total of 98 patients with chronic urticaria who were admitted to our hospital from July 2021 to March 2022 were selected and divided into random odd-even numbers. The odd numbers were included in the observation group, with a total of 49 cases, and they were treated with Fangfeng Tongsheng granule combined with levocetirizine; the even numbers were included in the control group, with a total of 49 cases and were treated with levocetirizine alone. The two groups of patients were treated continuously for 4 weeks, and the clinical efficacy of the two groups was observed. Before treatment, 2 weeks and 4 weeks after treatment, evaluate the clinical symptom scores of patients such as itching, flushing, wheal, edema, observe the improvement of clinical symptoms of patients, and the changes in Dermatology Life Quality Index (DLQI). Serum complement C3, C4, T lymphocyte subsets CD3 +, CD4 +, CD8 + levels and CD4 +/CD8 + ratio, IL-4, IgE, and IFN-γ levels and the occurrence of adverse reactions in the two groups were calculated and observed. All patients were followed up for 2 months after treatment to observe the recurrence of patients. Results: The scores of clinical symptoms such as wheal, itching, flushing, edema, and attack frequency in the observation group at each time point after treatment were lower than those in the control group (F times were 725.365, 851.521, 936.411, 3943.136, and 2226.147, all P < 0.05 (F between-group were 40.642, 102.124, 188.523, 259.291, and 23.92, P < 0.05); the total effective rate of the observation group was 93.88% (46/49), which was significantly higher than that of the control group, 73.47% (36/49) (χ 2 = 7.470, P=0.006). The DLQI scores of the observation group at each time point after treatment were lower than those of the control group (F time was 282.214, P < 0.05; F between-group was 6.546, P < 0.05). There was no significant difference in serum C4 levels between the two groups at each time point (F time was 1.225, P > 0.05; F between-group was 0.408, P > 0.05); serum complement C3, CD3 +, and CD4 +/the ratio of CD8 + and IFN-γ were higher than those in the control group (F time was 407.352, 107.823, 32.941, and 2354.147, P < 0.05; F between-group was 40.941, 24.710, 54.982, and 264.921, P < 0.05); the observation group at each time after treatment the levels of IgE and IL-4 were lower than those of the control group (F time were 373.124 and 395.612, P < 0.05; F between-group were 21.802 and 62.591, P < 0.05). The incidence of adverse reactions in the observation group was 12.24% (6/49) compared with 10.20% (5/49) in the control group, which had no significant difference (χ 2 = 0.102, P=0.749). Both groups were followed up for 2 months after treatment. The recurrence rate in the observation group was 12.24% (6/49), which was lower than that in the control group, which was 32.65% (16/49) (χ 2 = 5.861, P=0.015). Conclusion: The application of Fangfeng Tongsheng granules combined with levocetirizine in patients with chronic urticaria can effectively improve the clinical symptoms of patients, improve clinical efficacy, reduce the impact of the disease on life, improve the immune status of patients, and reduce the risk of recurrence.

Transcriptome Atlas of 16 Donkey Tissues.

Donkeys (Equus asinus) are important livestock with great economic value in meat, skin, and milk production. However, a lack of knowledge of the transcriptome landscape across a wide range of donkey tissues limits genetic selective breeding and conservation. Here we used transcriptomics to describe the transcriptome landscape, classify the tissue-specific gene expression across all primary donkey tissues, and present supplementary analyses on the protein level of additional donkey milk samples. Overall, 16,013 protein-coding genes and 21,983 transcripts were mapped to the reference genome, including 6,778 ubiquitously expressed genes and 2,601 tissue-enriched genes. Functional analysis revealed that the function of the tissue-enriched genes was highly tissue specific. Tissue-elevated genes that could be associated with unique phenotypes in donkey were analyzed. The results showed that, compared with those in human and other livestock, the lysozyme gene in donkey breast was specifically and highly expressed. The calcium-binding lysozyme, encoded by the lysozyme gene, was also detected in high amounts in donkey milk. Given those intact lysozyme genes that predict potentially functional calcium-binding lysozyme found in only a few species (e.g., donkey and horse), the high expression of the lysozyme gene in donkey breast may contribute to the high lysozyme content in donkey milk. Furthermore, 71% of the proteins in donkey milk overlapped with human milk protein, higher than the overlapping rates of bovine, sheep, and swine with humans. The donkey transcriptomic resource contributes to the available genomic resources to interpret the molecular mechanisms underlying phenotype traits.

Green and enhanced extraction of coumarins from Cortex Fraxini by ultrasound-assisted deep eutectic solvent extraction.

Green and enhanced extraction of bioactive ingredients from medicinal plants has become a hot research field, and deep eutectic solvents have been considered as a novel kind of sustainable solvents in the extraction process. In this study, hydrogen bond acceptor (choline chloride, etc.) and hydrogen bond donor (l-malic acid, etc.) were used to prepare different kinds of deep eutectic solvents to extract coumarins from Cortex Fraxini. The extraction conditions, including the composition and moisture content of deep eutectic solvents, extraction time, and liquid-solid ratio, were systematically optimized basing on the extraction yield of coumarins. To further investigate the extraction mechanism, Fourier transform infrared spectroscopy was performed, and the microstructures of Cortex Fraxini powders were observed before and after extraction using scanning electron microscope. Results showed that the novel ultrasound-assisted extraction with conditions of deep eutectic solvent containing betaine/glycerin (1:3), aqueous solution (20%), solid-liquid ratio (15 mg/mL), and extraction time (30 min) exhibited the best extraction yields for the four target coumarins and much better extraction efficiency than with conventional solvent extractions. This suggests that the new ultrasound-assisted deep eutectic solvent extraction could be used as a green and high-efficient approach for extraction of the main coumarins from Cortex Fraxini.

Pancreatic lipase inhibitory constituents from Fructus Psoraleae.

Pancreatic lipase (PL), a crucial enzyme in the digestive system of mammals, has been proven as a therapeutic target to prevent and treat obesity. The purpose of this study is to evaluate and characterize the PL inhibition activities of the major constituents from Fructus Psoraleae (FP), one of the most frequently used Chinese herbs with lipid-lowering activity. To this end, a total of eleven major constituents isolated from Fructus Psoraleae have been obtained and their inhibition potentials against PL have been assayed by a fluorescence-based assay. Among all tested compounds, isobavachalcone, bavachalcone and corylifol A displayed strong inhibition on PL (IC50 < 10 µmol·L-1). Inhibition kinetic analyses demonstrated that isobavachalcone, bavachalcone and corylifol A acted as mixed inhibitors against PL-mediated 4-methylumbelliferyl oleate (4-MUO) hydrolysis, with the Ki values of 1.61, 3.77 and 10.16 µmol·L-1, respectively. Furthermore, docking simulations indicated that two chalcones (isobavachalcone and bavachalcone) could interact with the key residues located in the catalytic cavity of PL via hydrogen binding and hydrophobic interactions. Collectively, these finding provided solid evidence to support that Fructus Psoraleae contained bioactive compounds with lipid-lowering effects via targeting PL, and also suggested that the chalcones in Fructus Psoraleae could be used as ideal leading compounds to develop novel PL inhibitors.

QuEChERS pretreatment combined with high-performance liquid chromatography-tandem mass spectrometry for determination of aristolochic acids I and II in Chinese herbal patent medicines.

Aristolochic acid I and II (AA I and II), a kind of nephrotoxic and carcinogenic compound, are widely added in Chinese herbal patent medicines though they have been banned due to their toxicity. However, the traditional sample pre-treatment combined with the LC-MS analysis system is not effective to determine AAs in such complicated patent medicines. The QuEChERS pretreatment method possesses some merits such as being quick and effective. In this work, the modified QuEChERS method was first used to determine AA I and II in Chinese herbal patent medicines combined with the HPLC-MS/MS analysis system. Extraction and removal of target analytes from powder, tablet, and capsule samples were conducted using the modified QuEChERS pretreatment. The liquid extracts of Chinese herbal patent medicines could be analyzed directly. The method optimization results show that average recoveries ranged from 96.6% to 110.3% with relative standard deviations ranging from 4.2% to 13.0%. The quantization limits of the three selected matrices are estimated as follows (AA I/II): 2.8/6.5 ng mL-l in liquid herbal extract, 6.5/12.5 ng g-1 in tablets, and 22.1/42.1 ng g-1 in capsules. This method was conducted to investigate the presence of AAs, which are a type of nephrotoxic and carcinogenic carboxylic acid, in 30 herbal products sold through the Internet in China. AA I and II were detected in 53% and 20%, respectively, of tested samples.

Correction: QuEChERS pretreatment combined with high-performance liquid chromatography-tandem mass spectrometry for determination of aristolochic acids I and II in Chinese herbal patent medicines.

[This corrects the article DOI: 10.1039/D0RA03200J.].

Discovery of natural pentacyclic triterpenoids as potent and selective inhibitors against human carboxylesterase 1.

Human carboxylesterase 1 (CES1), primarily expressed in the liver and adipocytes, is responsible for the hydrolysis of endogenous esters (such as cholesteryl esters and triacylglycerols) and the metabolism of xenobiotic esters (such as clopidogrel and oseltamivir), thus participates in physiological and pathological processes. In this study, a series of natural pentacyclic triterpenoids were collected and their inhibitory effects against CES1 and CES2 were assayed using D-luciferin methyl ester (DME) and N-(2-butyl-1,3-dioxo-2,3-dihydro-1H-benzo[de] isoquinolin- 6-yl)- 2-chloroacetamide (NCEN) as specific optical substrate for CES1, and CES2, respectively. To this end, betulinic acid (BA) was found with strong inhibitory effect on CES1 (IC50, 15 nM) and relative high selectivity over CES2 (>2400-fold). Primary structure-activity relationships (SAR) analysis and docking simulations revealed that the carboxyl group at the C-28 site of BA is very essential for CES1 inhibition. The inhibition kinetic analyses demonstrated that BA was a potent competitive inhibitor against CES1-mediated DME hydrolysis. Further investigation on the inhibitory effect of BA in living cells (HepG2) based assays demonstrated that BA displayed potent inhibitory effects on intracellular CES1 activities, with the low IC50 value of 1.30 µM. These results demonstrated that BA is potent and highly selective CES1 inhibitor, which might be used as the promising tool for exploring the biological functions of CES1 in complex biological systems.

Predictive metabolic signatures for the occurrence and development of diabetic nephropathy and the intervention of Ginkgo biloba leaves extract based on gas or liquid chromatography with mass spectrometry.

Diabetic nephropathy (DN) is one of the leading causes of death in diabetes mellitus (DM). Early warning and therapy has significant clinical value for DN. This research sought to find biomarkers to predict the occurrence and development of DN and the intervention of Ginkgo biloba leaves extract (GBE) by quantifying fatty acids, amino acids, and nucleosides and nucleobases in rat plasma. Samples were respectively collected at the weekend of 5-10 weeks after diabetic rats induced by streptozotocin were defined. Plasma fasting blood-glucose, kidney index, blood urea nitrogen, creatinine, urine albumin excretion and ultrastructural morphology of kidney were measured or observed. Fatty acids, amino acids and nucleosides and nucleobases in rat plasma were analyzed by gas chromatography or liquid phase chromatography and mass spectrometry, respectively. From the biochemical index and morphological change of kidney, the rats from the 5th to 7th week were in the stage of DM while from the begin of 8th week the rats were suggested in the early stage of DN. The results of quantitative metabolomics showed that 16 differential metabolites were related to the progression of DN, and oleic acid, glutamate and guanosine might be the potential biomarkers of kidney injury. 14 differential metabolites were related to GBE against the progression of DN, while oleic acid and glutamate might be the potential biomarkers of GBE against kidney injury. Those findings potentially promote the understanding of the pathogenic progression of DN and reveal the therapeutic mechanism of GBE against DN.

Process optimization of spray-dried fanhuncaoin powder for pulmonary drug delivery and its pharmacokinetic evaluation in rats.

The optimization of process parameters of spray-dried powder containing fanhuncaoin, a newly discovered anti-inflammatorily active phenolic acid isolated from Chinese herb, was conducted using response surface methodology (RSM). The experimental results were fitted into partial cubic polynomial model to describe and predict the response quality in terms of the final angle of repose, aerodynamic diameter, respirable fraction (RF), and yield. The recommended optimum spray-drying parameters for the development of fanhuncaoin powder with optimum quality were 110 °C inlet temperature, 0.50 m3/min aspiration speed, and 7.95 ml/min feed flow rate. The obtained optimum process parameters were employed for the production of spray-dried fanhuncaoin powder and to check the validity of the partial cubic model. Small and insignificant deviations were found between the predicted values and the experimental ones, showing the efficiency of the model in predicting the quality attributes of fanhuncaoin powder. The optimized powder was further examined for its pharmacokinetic properties in rats. A UPLC/MS assay was used to determine plasma fanhuncaoin concentration. Statistical analysis demonstrated that there was no significant difference in the t1/2 and dose-normalized Cmax and AUC as well as other pharmacokinetic parameters between the groups dosed differently following intratracheal administration (p > .05), indicating that fanhuncaoin followed linear kinetics. The pharmacokinetic parameters of fanhuncaoin after intratracheal administration differed significantly from the ones observed after intravenous administration (p < .05). The lower values of Cmax and AUC(0-∞) obtained following intratracheal administration may lead to effective drug concentrations at the target site with minimal systemic bioavailability and side effects.

Investigation of the preventive effect of Sijunzi decoction on mitomycin C-induced immunotoxicity in rats by 1H NMR and MS-based untargeted metabolomic analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Sijunzi decoction (SJZD) is a well known traditional Chinese prescription used for the treatment of gastrointestinal disorders and immunity enhancement. It has been found to indeed improve life quality of chemotherapy patients and extensive used in clinical conbined with chemotherapeutics for the treatment of cancer. AIM OF THE STUDY: The aim of this study was to investigate the preventive effect of the immunotoxicity of SJZD on mitomycin C (MMC) and the metabolic mechanism of action. MATERIALS AND METHODS: NMR and MS-based metabolomics approaches were combined for monitoring MMC-induced immunotoxicity and the protective effect of SJZD. Body weight change and mortality, histopathological observations and relative viscera weight determinations of spleen and thymus, sternum micronucleus assay and hematological analysis were used to confirm the immunotoxicity and attenuation effects. An OPLS-DA approach was used to screen potential biomarkers of immunotoxicity and the MetaboAnalyst and KEGG PATHWAY Database were used to investigate the metabolic pathways. RESULTS: 8 biomarkers in plasma samples, 19 in urine samples and 10 in spleen samples were identified as being primarily involved in amino acid metabolism, carbohydrate metabolism and lipid metabolism. The most critical pathway was alanine, aspartate and glutamate metabolism. CONCLUSIONS: The variations in biomarkers revealed the preventive effect of the immunotoxicity of SJZD on MMC and significant for speculating the possible metabolic mechanism.

Plant Uptake and Metabolism of Nitrofuran Antibiotics in Spring Onion Grown in Nitrofuran-Contaminated Soil.

Environmental pollution caused by the discharge of mutagenic and carcinogenic nitrofurans to the aquatic and soil environment is an emerging public health concern because of the potential in producing drug-resistant microbes and being uptaken by food crops. Using liquid chromatography-tandem mass spectrometry analysis and with spring onion (Allium wakegi Araki) as the plant model, we investigated in this study the plant uptake and accumulation of nitrofuran from a contaminated environment. Our study revealed for the first time high uptake and accumulation rates of nitrofuran in the edible parts of the food crop. Furthermore, results indicated highly efficient plant metabolism of the absorbed nitrofuran within the plant, leading to the formation of genotoxic hydrazine-containing metabolites. The results from this study may disclose a previously unidentified human exposure pathway through contaminated food crops.

Comparative pharmacokinetic study of the main components of cortex fraxini after oral administration in normal and hyperuricemic rats.

Cortex Fraxini is an important traditional Chinese herbal medicine used for the treatment of gout and hyperuricemia. An efficient and rapid ultra-performance liquid chromatography mass spectrometry method was developed and validated for simultaneous quantitation of six coumarins (aesculin, fraxin, aesculetin, fraxetin, sopoletin and 7-hydroxycoumarin) in normal and hyperuricemic rats plasma after oral administration of Cortex Fraxini. The method could successfully be applied for pharmacokinetics studies. The pharmacokinetic behavior of six coumarins in normal and hyperuricemia rats plasma was determined. Results showed that, for some of analytes, the pharmacokinetic parameters (AUC0-t , AUC0-∞ , Cmax , Tmax and CL) were significantly different between normal and hyperuricemic rats. The different pharmacokinetic parameters might result from renal impairment or a change of metabolic enzymes in the pathological state. The pharmacokinetic study in pathological state could provide more useful information to guide the clinical use of traditional Chinese herbal medicine.

Rapid characterization of the chemical constituents of Cortex Fraxini by homogenate extraction followed by UHPLC coupled with Fourier transform ion cyclotron resonance mass spectrometry and GC-MS.

Cortex Fraxini is an important traditional Chinese medicine. In this work, a rapid and reliable homogenate extraction method was applied for the fast extraction for Cortex Fraxini, and the method was optimized by response surface methodology. Ultra high performance liquid chromatography combined with Fourier transform ion cyclotron resonance mass spectrometry and gas chromatography with mass spectrometry were established for the separation and characterization of the constituents of Cortex Fraxini. Liquid chromatography separation was conducted on a C18 column (150 mm × 2.1 mm, 1.8 µm), and gas chromatography separation was performed on a capillary with a 5% phenyl-methylpolysiloxane stationary phase (30 m × 0.25 mm × 0.25 mm) by injection of silylated samples. According to the results, 33 chemical compounds were characterized by liquid chromatography with mass spectrometry, and 11 chemical compounds were characterized by gas chromatography with mass spectrometry, and coumarins were the major components characterized by both gas chromatography with mass spectrometry and liquid chromatography with mass spectrometry. The proposed homogenate extraction was an efficient and rapid method, and coumarins, phenylethanoid glycosides, iridoid glycosides, phenylpropanoids, and lignans were the main constituents of Cortex Fraxini. This work laid the foundation for further study of Cortex Fraxini and will be helpful for the rapid extraction and characterization of ingredients in other traditional Chinese medicines.

Profiling analysis of amino acids from hyperlipidaemic rats treated with Gynostemma pentaphyllum and atorvastatin.

UNLABELLED: Context Gynostemma pentaphyllum (Thunb.) Makino has been used in traditional medicine for the treatment of hyperlipidaemic with a long history. OBJECTIVE: The objective of this study was to evaluate the influence of Gynostemma pentaphyllum (GP) and atorvastatin on amino acids from the plasma and liver tissue of hyperlipidaemic rats. Materials and methods The rats were fed a high-fat diet continuously for 11 weeks for the construction of hyperlipidaemic model. The hyperlipidaemic rats were treated with Gynostemma pentaphyllum (120 mg/kg) and atorvastatin (1.8 mg/kg) for 4 weeks, and the rats were intragastric administration one time every day. Chromatographic separation was performed on a Shim-pack XR-ODSIII C18 analytical column (75 mm × 2.0 mm i.d., 1.6 µm, Shmadazu Corp., Tokyo, Japan). The biomarkers of amino acids were identified by principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA). Results After feeding with a high-fat diet, the TC and LDL-C values of the hyperlipidaemic mode rats increased dramatically (p < 0.01). The established method allowed a target analysis of 12 kinds of amino acids. PCA studies showed that the plasma amino acids had not returned to normal after GP treatment, but those had recovered slightly after atorvastatin treatment. GP has almost no impact on the metabolism of amino acids, while atorvastatin can modify the metabolism of amino acids via self-regulatory mechanisms. Discussion and conclusion UPLC/DAD combined with SCX-SPE can be successfully used for profiling analysis of amino acids. By the comparison of biomarkers following treatment with GP and atorvastatin, the influence of the two drugs on biomarkers is revealed.

(1)H NMR and MS based metabolomics study of the therapeutic effect of Cortex Fraxini on hyperuricemic rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Cortex Fraxini (CF) is an important traditional Chinese herbal medicine used for the treatment of gout and hyperuricemia. AIM OF THE STUDY: The aim of this study was to evaluate the anti-hyperuricemic effect of CF on hyperuricemic rats and to investigate its mechanism of action. MATERIALS AND METHODS: Metabolomics based on NMR and MS was used to study the therapeutic effect of CF on hyperuricemic rats. Plasma determination of uric acid (UA) showed that CF treatment markedly improved the UA level. Subsequently, metabolomics analysis was conducted using samples of plasma, kidney and urine, and orthogonal partial least squares-discriminant analysis (OPLS-DA) combined with principal component analysis (PCA) were used to detect potential biomarkers. RESULTS: A total of 26 biomarkers were identified as being primarily involved in amino acid metabolism, lipid metabolism, purine metabolism, amino acid metabolism and carbohydrate metabolism, and hyperuricemia can disturb the balance of many of these metabolic pathways in vivo. CONCLUSIONS: The variations in biomarkers revealed the therapeutic mechanism of CF, and a number of these biomarkers are not only significant for early diagnosis but also for predicting hyperuricemia.