Xiebai San alleviates acute lung injury by inhibiting the phosphorylation of the ERK/Stat3 pathway and regulating multiple metabolisms.

BACKGROUND: Acute lung injury (ALI) often leads to serious respiratory diseases with high incidence rates and mortality. For centuries, Xiebai San (XBS) has been a classical traditional Chinese medicine (TCM) about respiratory illness such as pneumonia in children. However, the related mechanism of XBS against ALI remains indistinct. PURPOSE: To reveal specific targets of XBS in lipopolysaccharide (LPS)-induced ALI mice using integrated pharmacology. STUDY DESIGN: The integrated method was to expound mechanism and targets of XBS inhibited ALI. METHODS: The primary components in XBS were identified by ultra high performance liquid chromatography-quadrupole time of flight-mass spectrometry (UHPLC-QTOF-MS). The potential drug targets were established using network pharmacology. The anti-ALI effect of XBS was evaluated in mice. Additionally, therapeutic targets were screened by integrating metabolome and transcriptome and verified in lung tissue. RESULTS: In total, 163 chemical components were identified in XBS, and a network of "3 drugs-18 components-86 targets" for XBS against ALI was constructed. In ALI mice, XBS alleviated lung inflammation by decreasing permeation and expression of neutrophils, tumor necrosis factor α (TNF-α), interleukin-6 (IL-6), and interleukin-1ß (IL-1ß) in bronchoalveolar lavage fluid (BALF), serum, and lung tissue. Next, the transcriptome of lung tissue was analyzed and enriched, indicating the importance of mitogen-activated protein kinase (MAPK), Janus kinase-signal transducer and activator of transcription (JAK-STAT), and others, which was consistent with network pharmacology prediction. Also, western blotting and immunohistochemistry results showed that XBS was against ALI mainly by inhibiting extracellular signal regulated kinase (ERK) and signal transducer and activator of transcription 3 (Stat3) phosphorylation. In addition, the metabolome of lung tissue revealed that XBS mainly regulated pathways involved in arachidonic acid, glycerophospholipid, and tryptophan metabolisms. The expression levels of leukotriene, phosphatidylcholine, kynurenine, and others were also verified. CONCLUSION: XBS alleviated inflammation of ALI by inhibiting the phosphorylation of the ERK/Stat3 pathway and regulating arachidonic acid, glycerophospholipid, and tryptophan metabolisms. This study will guide clinical precision medicine and promote modernization of XBS.

Ginkgolides with anti-PAF activity from Ginkgo biloba L.

Four undescribed ginkgolides, including two rare sesquiterpene ginkgolides (compounds 1 and 2) and two diterpenoid ginkgolides (compounds 3 and 4), were isolated from Ginkgo biloba L. The structures of these four ginkgolides were identified based on extensive spectroscopic analysis, DP4+ probability analysis and X-ray diffraction. Compounds 1 and 2 exhibited excellent antiplatelet aggregation activities with IC50 values of 1.20 ± 0.25 and 4.11 ± 0.34 µM, respectively.

Pharmacodynamics and Mechanism of Astragali Radix and Anemarrhenae Rhizoma in Treating Chronic Heart Failure by Inhibiting Complement Activation.

Astragali radix (AR) and anemarrhenae rhizoma (AAR) are used clinically in Chinese medicine for the treatment of chronic heart failure (CHF), but the exact therapeutic mechanism is unclear. In this study, a total of 60 male C57BL/6 mice were divided into 5 groups, namely sham, model, AR, AAR, and AR-AAR. In the sham group, the chest was opened without ligation. In the other groups, the chest was opened and the transverse aorta was ligated to construct the transverse aortic constriction model. After 8 weeks of feeding, mice were given medicines by gavage for 4 weeks. Left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) were detected by echocardiography. Heart weight index (HWI) and wheat germ agglutinin staining were used to evaluate cardiac hypertrophy. Hematoxylin-eosin staining was used to observe the pathological morphology of myocardial tissue. Masson staining was used to evaluate myocardial fibrosis. The content of serum brain natriuretic peptide (BNP) was detected by enzyme-linked immunosorbent assay kit. The content of serum immunoglobulin G (IgG) was detected by immunoturbidimetry. The mechanism of AR-AAR in the treatment of CHF was explored by proteomics. Western blot was used to detect the protein expressions of complement component 1s (C1s), complement component 9 (C9), and terminal complement complex 5b-9 (C5b-9). The results show that AR-AAR inhibits the expression of complement proteins C1s, C9, and C5b-9 by inhibiting the production of IgG antibodies from B cell activation, which further inhibits the complement activation, attenuates myocardial fibrosis, reduces HWI and cardiomyocyte cross-sectional area, improves cardiomyocyte injury, reduces serum BNP release, elevates LVEF and LVFS, improves cardiac function, and exerts myocardial protection.

In vitro digestion and fecal fermentation of selenocompounds: impact on gut microbiota, antioxidant activity, and short-chain fatty acids.

Selenium bioavailability is critically influenced by gut microbiota, yet the interaction dynamics with selenocompounds remain unexplored. Our study found that L-Selenomethionine (SeMet) and Se-(Methyl)seleno-L-cysteine (MeSeCys) maintained stability during in vitro gastrointestinal digestion. In contrast, Selenite and L-Selenocystine (SeCys2) were degraded by approximately 13% and 35%. Intriguingly, gut microflora transformed MeSeCys, SeCys2, and Selenite into SeMet. Moreover, when SeCys2 and Selenite incubated with gut microbiota, they produced red selenium nanoparticles with diameters ranging between 100 and 400 nm and boosted glutathione peroxidase activity. These changes were positively associated with an increased relative abundance of unclassified_g__Blautia (Family Lachnospiraceae), Erysipelotrichaceae_UCG-003 (Family Erysipelatoclostridiaceae), and uncultured_bacterium_g__Subdoligranulum (Family Ruminococcaceae). Our findings implied that differential microbial sensitivities to selenocompounds, potentially attributable to their distinct mechanisms governing selenium uptake, storage, utilization, and excretion.

New Glycerolipids from the Traditional Chinese Medicine of Syngnathus acus (Hai-Long).

Two new glycerolipids, syngaculipids A and B (1 and 2), one first naturally occurring metabolite (8), together with five known compounds (3-7) were isolated from the AcOEt fraction of Syngnathus acus L. (Hai-Long). Their structures were elucidated by comprehensive spectral analyses involving UV, IR, MS, 1D and 2D NMR spectra and ECD calculations. All the isolated compounds were evaluated for their cytotoxicity against A549 and HCT-116 cell lines. Compound 8 exhibited moderate cytotoxicity with IC50 values of 34.5 and 38.9 µM on the A549 and HCT-116 cell lines, respectively.

[Improvement of high-quality evaluation criteria of Chinese patent medicines based on whole process control].

Chinese patent medicines(CPMs) are unique therapeutic drugs in China. Establishing and improving the evaluation criteria is an important measure to promote the high-quality development of CPMs. Based on the "evaluation criteria of high-grade CPMs with quality as the core index" established by our group in 2018, the "high-quality evaluation criteria for CPMs based on whole process control" was proposed in the present study in 2022. The scope of application and basic principles of the new criteria were clarified. A quality evaluation scoring table was established in the new criteria, including five parts: raw material selection, production process, quality control, efficacy evaluation, and brand building. The technical evaluation indexes involved have increased from 20% in the original criteria to 70% in the new criteria, and efficacy evaluation has been added in the new criteria. The subjective evaluation indicators account for a large proportion in the original criteria, which is prone to bias. The improved criteria overcome this shortcoming. It is expected that the new criteria as a basis can play a better role in the selection of high-quality products of CPMs, guide enterprises and institutions to participate actively in the evaluation and research of high-quality CPMs, and promote the high-quality development of CPMs.

[Advantages and disadvantages of production quality of Fufang Danshen Prescription based on high-quality Chinese patent medicine evaluation criteria].

The big brand of Chinese patent medicine, Fufang Danshen Prescription(FDP), effective in promoting blood circulation, resolving blood stasis, regulating qi, and relieving pain, is wide in clinical application and diverse in dosage forms and products, but its quality and price are uneven, which causes problems for doctors and patients. To clarify the key links and weakness of quality control leading to the quality difference of FDP products, the present study carried out a comprehensive evaluation of the whole production cycle of FDP based on the "high-quality Chinese patent medicine evaluation criteria" and analyzed the advantages and disadvantages of production and quality of different FDP products according to scores. The results showed that the scores of various products in the "raw materials selection" varied greatly. The highest score(S1) and the lowest score(S2) differed by more than 3 times, indicating that different manufacturers had inconsistent requirements for the selection of raw materials, leading to fundamental differences in the quality of raw materials. The scores in the "production process" varied slightly, with an average score of 66.8%. The manufacturer S8 obtained the highest score(84.0%), which indicated the emergence of intelligent manufacturing production. The scores(with the average score of 44.0%) in the "quality control" were lower than those of the previous two items, which was attributed to the fact that most FDP products only met the "qualified" benchmark required by the 2020 edition of Chinese Pharmacopoeia, and their consistency and high quality were both uncontrollable. The scores in the "post-marketing research" were the lowest(with an average score of 28.5%), and most manufacturers were scored 0, which reflected little attention paid. Only a few brand manufacturers were scored acceptably and they were willing to carry out relevant research on post-marketing evaluation. The evaluation results demonstrated the key links and weakness leading to the production and quality differences of FDP from different manufacturers. It is expected to improve the quality of FDP, promote the formation of the "high quality and good price" mechanism, and provide information for the centralized procurement of governments.

Discovery of unreported ginkgolides of anti-PAF activity using characteristic ion and neutral loss recognition strategy in Ginkgo biloba L.

Ginkgolides are the most important bioactive components of Ginkgo biloba L, of which ginkgolide B has been successfully developed and marketed as a drug. The reported ginkgolides are very rare and exhibit a complex matrix due to the chemodiversity of Ginkgo biloba L. Herein, the global profile of characteristic ion and neutral loss recognition strategy were used for to discover eight undescribed ginkgolides, very rare cyclohexane ginkgolides R-V, ginkgolides D-F, and eight known ginkgolides. These ginkgolides were target isolated and identified using high-resolution mass spectrometry, nuclear magnetic resonance spectroscopy, and X-ray crystallography. The undescribed and known ginkgolides exhibited antiplatelet aggregation activities. In particular, compounds U and D had IC50 values of 2.20 ± 0.15 and 6.50 ± 0.87 µM, respectively. This study has enriched the known structural diversity of ginkgolides and extended the application of mass spectrometry to the global profiling of natural products present in Ginkgo biloba L. Moreover, it could help chemists rapidly discover unreported compounds from a complex matrix.

Comprehensive quality consistency evaluation strategy and analysis of compound danshen tablet.

The compositions of traditional Chinese medicines are extremely complex,as a result, exploring consistent quality is demanded and challenging. Quality consistency of products obtained from the same manufacturer has received little attention. The strategy of quality consistency evaluation (QCE) has been proposed as a novel method for quality control of Traditional Chinese Medicine Patent Prescription (TCMPP). This study aimed to establish a comprehensive QCE strategy for Compound Danshen Tablet (CDT). High Performance Liquid Chromatography-Diode Array Detector and Gas Chromatography-Mass Spectrometry were separately applied to determinate the content of seven and two index components, which representing the quality actuality of different raw medicines. The dissolution test was designed to obtain the dissolution ratios of CDT samples. QCE can provide the intra-batch content consistency difference (PA), inter-batch content consistency difference (PB), and dissolution ratio consistency difference (PR) values. The consistency of CDT samples from 15 different manufacturers (75 batches) was evaluated by principal component analysis (PCA), which showed that the total content (nine index components) of the 75 batches of samples obtained from 15 manufacturers ranged from 22.11 to 38.45 mg·tablet-1. The dissolution ratios ranged from 74.8% to 116.4%. The PA values of 15 manufacturers ranged from 2.4% to 12.2%, and the PB (11.1-45.1%) values were higher than the PA values. The PR values reflecting the various dissolution ratios in vitro ranged from 8.1% to 57.5%. The three consistency factors were ranked by PCA, and products of the 15 manufacturers were classified into three categories. The PA, PB, and PR values provided a comprehensive and effective approach for monitoring the quality consistency of CDT and can serve as an example of QCE for other TCMPP.

ß-Caryophyllene suppresses ferroptosis induced by cerebral ischemia reperfusion via activation of the NRF2/HO-1 signaling pathway in MCAO/R rats.

BACKGROUND: Ischemic stroke is a complex brain disease regulated by several cell death processes, including apoptosis, autophagy, and ferroptosis. ß-Caryophyllene (BCP), a natural bicyclic sesquiterpene abundantly found in essential oils, has been demonstrated to have potential pharmacological benefits in many diseases, including ischemic stroke. PURPOSE: This research was to determine the existence of ferroptosis in the pathogenesis of acute ischemic stroke and investigate whether BCP can inhibit ferroptosis to improve cerebral ischemia injury by activating the NRF2/HO-1 signaling pathway in rats. METHODS: First, we verified ferroptosis by assessing proferroptotic changes after middle cerebral artery occlusion reperfusion (MCAO/R), along with protein and lipid peroxidation levels. Then male rats were divided randomly into Sham, MCAO/R, ML385 (NRF2-specific inhibitor) and BCP groups. The effects of BCP on cerebral injury were detected by the modified neurological severity score, TTC staining, and hematoxylin-eosin staining. We conducted western blotting analyzes of proteins, including those involved in ferroptosis and related signaling pathways. To demonstrate the neuroprotective effect of BCP in vitro, primary astrocytes were pretreated with different concentrations of BCP (10, 20, and 40 µM) for 24 h before oxygen-glucose deprivation/re-oxygenation (ODG/R). RESULTS: We concluded that ferroptosis was engaged in the process of I/R-induced neurological damage, implying that this novel type of cell death might provide new therapeutic options for the clinical treatment of ischemic stroke. In vivo study proved that BCP improved neurological scores, infarct volume, and pathological features after MCAO/R. We demonstrated that BCP evidently enhanced NRF2 nuclear translocation, activated the NRF2/HO-1 pathway, which protected against ferroptosis. In vitro investigation revealed the same results. BCP decreased OGD/R-induced ROS generation and iron accumulation. Furthermore, the neuroprotective effects of BCP were reversed by the NRF2 inhibitor ML385. CONCLUSION: Our results indicated the critical role of ferroptosis in cerebral I/R injury. For the first time, we showed that the significant neuroprotective effects of BCP in attenuating ischemic stroke injury are correlated with ferroptosis regulation, and its mechanism is associated with activation of the NRF2/HO-1 axis.

Using recycled coffee grounds for the synthesis of ZIF-8@BC to remove Congo red in water.

Around 6.6 million tons of spent coffee is produced per year, resulting in resources loss and potential environmental risks. Hence, a green technique is required to reuse the spent coffee grains. In this study, coffee grounds were burnt at 900 °C to generate the biochar (BC) for the synthesis of the porous adsorbent (ZIF-8 @BC) by growing ZIF-8 on the surface of BC. We applied the well-prepared ZIF-8 @BC to remove Congo red (CR) in water. The maximum adsorption capacity of ZIF-8 @BC on Congo red in water was up to 1080.4 mg/g, which was significantly higher than that of many different types of BCs reported in previous studies. The reasons for its highly efficient adsorption of CR probably was attributed to metal ions and coordinatively unsaturated sites in the material. Also, BC enabled the less aggregation of ZIF-8 to provide sufficient specific surface area for CR adsorption. From the analysis of the pseudo-second-order kinetic model and Langmuir model, the adsorption of ZIF-8 @BC on CR was a homogeneously chemical adsorption process regulated by electrostatic interaction, π-π stacking and metal coordination.

Qualitative analysis and differentiation of ginkgo cultivars based on UHPLC-QTOF-MS/MS with the characteristic ion and neutral loss strategy combined with chemometric methods.

The identification of chemical constituents can assist the discovery of active ingredients and can differentiate herbs with multiple cultivars. In this study, a diagnostic ion and neutral loss filtering strategy was developed for the qualitative analysis of ginkgo leaf. The strategy is based on an ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry. A large number of (110) of GL compounds were identified, including 8 potentially novel compounds and 42 previously unreported GL constituents. Moreover, 64 available compounds in 48 GL cultivars were analyzed via a combined multicomponent quantitative analysis and statistical analysis. The distribution of the 64 compounds among different cultivars was clarified in a principal component analysis and hot map visualization. Via a variable-importance-for-prediction score analysis, ten main differential compounds were found among the different cultivars. Collectively, these results indicated the usefulness of our approach in chemical profiling and discrimination of herbs with multiple botanical origins. This strategy can also help chemists rapidly identify novel compounds from a complex matrix.

Network pharmacology and molecular docking reveal the effective substances and active mechanisms of Dalbergia Odoriferain protecting against ischemic stroke.

Dalbergia Odorifera (DO) has been widely used for the treatment of cardiovascular and cerebrovascular diseasesinclinical. However, the effective substances and possible mechanisms of DO are still unclear. In this study, network pharmacology and molecular docking were used toelucidate the effective substances and active mechanisms of DO in treating ischemic stroke (IS). 544 DO-related targets from 29 bioactive components and 344 IS-related targets were collected, among them, 71 overlapping common targets were got. Enrichment analysis showed that 12 components were the possible bioactive components in DO, which regulating 9 important signaling pathways in 3 biological processes including 'oxidative stress' (KEGG:04151, KEGG:04068, KEGG:04915), 'inflammatory response'(KEGG:04668, KEGG:04064) and 'vascular endothelial function regulation'(KEGG:04066, KEGG:04370). Among these, 5 bioactive components with degree≥20 among the 12 potential bioactive components were selected to be docked with the top5 core targets using AutodockVina software. According to the results of molecular docking, the binding sites of core target protein AKT1 and MOL002974, MOL002975, and MOL002914 were 9, 8, and 6, respectively, and they contained 2, 1, and 0 threonine residues, respectively. And some binding sites were consistent, which may be the reason for the similarities and differences between the docking results of the 3 core bioactive components. The results of in vitro experiments showed that OGD/R could inhibit cell survival and AKT phosphorylation which were reversed by the 3 core bioactive components. Among them, MOL002974 (butein) had a slightly better effect. Therefore, the protective effect of MOL002974 (butein) against cerebral ischemia was further evaluated in a rat model of middle cerebral artery occlusion (MCAO) by detecting neurological score, cerebral infarction volume and lactate dehydrogenase (LDH) level. The results indicated that MOL002974 (butein) could significantly improve the neurological score of rats, decrease cerebral infarction volume, and inhibit the level of LDH in the cerebral tissue and serum in a dose-dependent manner. In conclusion, network pharmacology and molecular docking predicate the possible effective substances and mechanisms of DO in treating IS. And the results are verified by the in vitro and in vivo experiments. This research reveals the possible effective substances from DO and its active mechanisms for treating IS and provides a new direction for the secondary development of DO for treating IS.

Incipient diploidization of the medicinal plant Perilla within 10,000 years.

Perilla is a young allotetraploid Lamiaceae species widely used in East Asia as herb and oil plant. Here, we report the high-quality, chromosome-scale genomes of the tetraploid (Perilla frutescens) and the AA diploid progenitor (Perilla citriodora). Comparative analyses suggest post Neolithic allotetraploidization within 10,000 years, and nucleotide mutation in tetraploid is 10% more than in diploid, both of which are dominated by G:C → A:T transitions. Incipient diploidization is characterized by balanced swaps of homeologous segments, and subsequent homeologous exchanges are enriched towards telomeres, with excess of replacements of AA genes by fractionated BB homeologs. Population analyses suggest that the crispa lines are close to the nascent tetraploid, and involvement of acyl-CoA: lysophosphatidylcholine acyltransferase gene for high α-linolenic acid content of seed oil is revealed by GWAS. These resources and findings provide insights into incipient diploidization and basis for breeding improvement of this medicinal plant.

Application of Multigroup Technology in Non-Small-Cell Lung Cancer with Qi Stagnation and Blood Stasis Syndrome.

OBJECTIVE: To explore the basic characteristics of intestinal flora, metabolomics, and proteomics of non-small cell lung cancer (NSCLC) in patients with Qi stagnation and blood stasis syndrome. METHODS: Twelve NSCLC patients with Qi stagnation and blood stasis syndrome were selected for the QZXY group and 15 healthy volunteers were selected for the control group. Fecal samples from the two groups were collected to evaluate intestinal microecology using the 16s rDNA technique. Serum samples were collected to compare the differences in metabolomics and proteomics between the two groups using liquid chromatography-mass spectrometry (LC-MS). Another 34 NSCLC patients with other syndromes were selected for the nQZXY group and their serum samples were collected. Metabolomics differences between the QZXY and nQZXY groups were compared using LC-MS, and four metabolites with the most obvious differences were selected for receiver operation characteristic curve representation. Finally, multigroup results were analyzed using the WGCNA software. RESULTS: There were two significantly different types of bacteria (Aerococcaceae and Abiotrophia), 11 different proteins (six upregulated and five downregulated), and 38 different metabolites (nine upregulated, 29 downregulated) between the QZXY and control groups. There was a correlation between differential bacteria, proteins, and metabolites. The conjoint analysis found that the different substances were related to MAPK, PI3K/Akt, Ras signaling pathway, cancer pathways, and cytokine-cytokine receptor interaction. There were four significant differences in metabolites (Pseudouridine, phenlacetyl-C0A, L-glutamic, and phospho-anandamide) between the QZXY and nQZXY groups. CONCLUSIONS: NSCLC with Qi stagnation and blood stasis syndrome had specific intestinal flora and protein and metabolites, which were closely related to the occurrence and development of tumors.

A simple strategy to monitor the temporal and spatial distribution of alkaloids in sacred lotus leaves.

Owing to the high degree of diversity of metabolite pools and complexity of spatial and temporal distributions within biological tissues, currently available methods for metabolite characterization face large challenges. In this study, the temporal and spatial distributions of the alkaloid components of the medicinal plant lotus (Nelumbo nucifera) were investigated over various growth phases. The results showed that alkaloid biosynthesis in lotus leaf is regulated by development and that there is maximum accumulation of alkaloids when the lotus leaf was completely expanded. Furthermore, alkaloid content tended to be stable in mature lotus leaves. However, there was significant variation in the alkaloid content of lotus leaves with different genotypes, suggesting that genetic background is an important factor that affects the temporal and spatial distributions of alkaloids in sacred lotus leaves. The dynamic contents of alkaloids during the growth and development of lotus leaves provide insight into basic biological differences when sampling.

The complete chloroplast genome sequence of Clematis Montana Buch.-Ham. (Ranunculaceae) and its phylogenetic analysis.

Clematis montana is a medicinal plant commonly used in southwest of China. The complete chloroplast (cp) genome sequence of C. montana was sequenced using the Illumina Hiseq 4000 platform. The cp genome of C. montana was 159,523 bp in length with 37.98% overall GC content. This circular molecule had a typical quadripartite structure containing a large single-copy (LSC) region of 79,385 bp, a small single-copy (SSC) region of 18,092 bp, and two inverted repeat (IR) regions of 31,023 bp. The cp genome contained 135 genes, including 91 protein-coding genes, 36 tRNA genes, and 8 rRNA genes. Phylogenetic analysis based on whole cp genome sequences showed that C. montana was closest to C. alternata.

The bioavailability and excretion of an antitussive compound IAsp-N-Glc in rats by validated UPLC-MS/MS methods.

IAsp-N-Glc is a potential antitussive agent that is first reported to be isolated from Ginkgo Semen, but the bioavailability and excretion of IAsp-N-Glc are unknown. Therefore, we carried out our study to obtain the bioavailability and excretion profiles of IAsp-N-Glc in rats. Rapid, specific, and reliable quantification methods for the measurement of IAsp-N-Glc in rat plasma and fecal samples by using ultra-high-performance liquid chromatography coupled with triple quadrupole mass spectrometry were developed and validated. A C18 column was used for the separation of IAsp-N-Glc and internal standards, and water (containing 0.1% formic acid) and acetonitrile were chosen as the mobile phase for the separation in the flow-gradient mode. In the ranges of 37.5-7500 ng/mL and 120-30000 ng/mL, the calibration curves of IAsp-N-Glc exhibited satisfactory linearity for plasma and fecal samples with each linear correlation coefficient higher than 0.99, respectively. The methods were reproducible and reliable. The analytes were stable, and no apparent matrix effects were observed. The bioanalytical methods were successfully used to study the pharmacokinetics and excretion of IAsp-N-Glc in rats. Oral administration of IAsp-N-Glc exhibited a low absolute oral bioavailability (1.83±0.09%), and 59.63±6.29% of IAsp-N-Glc was excreted in feces. This report is the first to describe the bioavailability and excretion of IAsp-N-Glc in rats and will lay the foundation for the in-depth study and drug development of IAsp-N-Glc.

Identification and quantification of target metabolites combined with transcriptome of two rheum species focused on anthraquinone and flavonoids biosynthesis.

Rheum emodi is a perennial herb and an important medicinal plant, with anthraquinones and flavonoids as its main bioactive compounds. However, there is little knowledge about the biosynthetic pathway of anthraquinones in rhubarbs. In this study, we qualitatively and quantitatively assessed 62 pharmacological metabolites in rhubarb using dynamic multiple reaction monitoring (dMRM) of triple-quadrupole mass spectrometry (QqQ-MS), including 21 anthraquinones, 17 flavonoids, 6 stilbenes, 12 gallate esters, 3 tannins, and 3 others. Besides, the metabolomics results showed significant differences among all the 60 metabolites, except for gallic acid and piceatannol-O-ß-glucoside. The combined transcriptome data of R. palmatum L. (RPL) and R. officinale Baill. (ROB) showed that 21,691 unigenes were annotated in the metabolic pathways. Taken together, 17 differentially expressed genes (DEGs) were associated with the anthraquinone biosynthetic pathway. Additionally, a significant correlation between anthraquinone peak intensity and DEG expression level existed, validating that DEGs contribute to the anthraquinone biosynthetic pathway. RT-qPCR results showed that the cluster-14354.38156 gene may catalyze the O-methylation of emodin to produce physcion. This study provides a useful resource for further studies on secondary metabolism in rhubarb and the combination analysis of transcriptome and metabolome, which can help with the discovery of enzyme genes involved in metabolite biosynthesis.

Quantification of 10 bioactive components of Yazhangsan in rat plasma by LC-MS/MS and its application.

Yazhangsan (YZS) is a common prescription for the treatment of cough and asthma caused by wind-cold. The purpose of this study was to investigate the pharmacokinetic profiles of 10 bioactive components in YZS. A simple, sensitive and reliable high-performance liquid chromatography coupled with a triple-quadruple mass spectrometry method (LC-MS/MS) was developed and fully validated in this study for the measurement of these 10 bioactive compounds in rat plasma. One-step protein precipitation method using methanol was applied to the treatment of rat plasma samples. Chromatographic separation was conducted on a C18 column by gradient elution, and water (containing 0.1% formic acid) and acetonitrile were chosen as the mobile phase. The analytes were quantified by using a mass spectrometer in multiple reaction monitoring scanning mode, and electrospray ionization was performed in positive and negative ion modes. The established method met the requirements for the quantification of these 10 bioactive compounds in biological samples, and it was successfully applied to the pharmacokinetic study of 10 components in rats after the intragastrical administration of YZS. This study will lay a foundation for the investigation of the mechanism of action of YZS and provide useful data for the rational use of YZS in clinical.