Exploration of material basis: Chemical composition profile, metabolic profile and pharmacokinetic characteristics in Xingbei Zhike granule.

HEADINGS ETHNOPHARMACOLOGICAL RELEVANCE: Xingbei Zhike granule (XBZK), a widely prescribed Chinese patent medicine, is known for its efficacy in clearing lung qi, relieving cough and reducing phlegm, as well as fever, dry and bitter taste, and irritability. Despite its clinical popularity, comprehensive investigations into its chemical composition, in vivo metabolism, and pharmacokinetic characteristics are limited. AIM OF THE STUDY: This study investigates the chemical composition, in vivo metabolism, and in vivo dynamics of XBZK to clarify its material basis and pharmacokinetic characteristics. MATERIALS AND METHODS: Ultra-high performance liquid chromatography with Orbitrap tandem mass spectrometry (UPLC-Orbitrap-MS) was used to determine the chemical composition and in vivo metabolic profile of XBZK. Additionally, UPLC with triple quadrupole mass spectrometry (UPLC-TQ-MS/MS) was performed to quantify its main components and evaluate its in vivo dynamics in rat plasma. RESULTS: In total, 57 components were identified in XBZK. Furthermore, 40 prototype components and 31 metabolites were detected in various biological matrices of rats, including plasma, tissues, bile, feces, and urine. After administration, the area under the curve (AUC) for ephedrine (Eph), pseudoephedrine (Peph), neotuberostemonine (Neo), amygdalin (Amy), and enoxolone (Eno) exhibited a strong linear relationship with the administered dose (r > 0.9) in all rats. And gender-related differences in the absorption of peiminine (Pmn), peimisine (Pms), and chrysin-7-O-glucuronide (Cog) were notable among rats, with male rats showing a dose-dependent pattern of absorption, while female rats exhibited minimal absorption. CONCLUSIONS: XBZK contains 57 components, primarily composed of flavonoids, alkaloids, and coumarins. The eight main components were rapidly absorbed and eliminated, with some, such as Eph, Peph, Neo, Amy and Eno, following a linear pharmacokinetic pattern. Furthermore, Pmn, Pms and Cog were well absorbed in male rats, showing a dose-dependent behavior.

Discovery of the material basis of Jiuwei Xifeng granules using pharmaco-chemistry and pharmacokinetics.

ETHNOPHARMACOLOGICAL RELEVANCE: Jiuwei Xifeng granules (JWXF) is primarily used for the treatment of Tourette syndrome (TS) with kidney-Yin deficiency and internal stirring of liver wind. However, few studies have focused on this issue. AIM OF THE STUDY: This study aimed to clarify chemical composition of JWXF using in vitro and in vivo pharmaco-chemistry and to provide a basis for the clinical use of JWXF using a strategy of pharmacokinetics. MATERIALS AND METHODS: In this study, the chemical constituents and in vivo metabolism of JWXF were evaluated using high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS/MS), and the time-dependent processes of the three main components in rats were detected using ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-QQQ-MS/MS). RESULTS: A total of 75 constituents were identified, including 22 alkaloids, 21 terpenes, 15 organic acids and their derivatives, and 17 other compounds. After administration, 12 compounds were identified in rat plasma, including 11 prototypes and one metabolite. Pharmacokinetic analysis showed that the effects of gentiopicroside, gastrodin, and sweroside in rats were dose-dependent when the dose of JWXF was 1-4 g/kg. They were rapidly absorbed and did not accumulate in the plasma after 7-day continuous intragastric administration. CONCLUSIONS: JWXF consists of 75 components, including alkaloids, terpenes, and organic acids. The three main compounds, gastrodin, gentiopicroside, and sweroside, undergo rapid absorption, elimination, and dose-dependent pharmacokinetics.

The tissue distribution of Jinzhen oral liquid in healthy and influenza mice.

Jinzhen oral liquid (JZOL) is widely used in China. However, its tissue distribution, a vital part of the efficacy substances research, has not been reported yet. This study characterized its chemical components and its prototypes and metabolites in mice, and investigated its tissue distribution in pathological and healthy mice. Several constituents were characterized, including 55 constituents in JZOL, 11 absorbed prototypes and six metabolites in plasma and tissues. The metabolic pathways were demethylation, dehydration and acetylation. A sensitive, accurate and stable quantitative method was established and applied to the tissue distribution. After administration of JZOL, these seven components were rapidly distributed to various tissues, mainly staying in the small intestine, and less distributed to lung, liver and kidney. Compared with healthy mice, the absorption of baicalin, wogonoside, rhein, glycyrrhizic acid and liquiritin apioside was reduced in influenza mice, but their elimination was slow. However, influenza infection had no obvious effect on the overall distribution of the most important components (baicalin, glycyrrhizic acid and wogonoside) in the plasma or small intestine, but obviously affected the distribution of baicalin in liver. In summary, seven components are rapidly distributed to various tissues, and influenza infection has certain influence on the tissue distribution of JZOL.

Mechanism of Jinzhen Oral Liquid against influenza-induced lung injury based on metabonomics and gut microbiome.

ETHNOPHARMACOLOGICAL RELEVANCE: Jinzhen Oral Liquid (JZOL) is a traditional Chinese patent medicine and widely used clinically, which consists of eight herbs including Bovis Calculus Atifactus, Fritillariae Ussuriensis Bulbus (Fritillaria ussuriensis Maxim.), Caprae Hircus Cornu, Rhei Radix et Rhizoma (Rheum palmatum L.), Scutellariae Radix (Scutellaria baicalensis Georgi), Glycyrrhizae Radix et Rhizoma (Glycyrrhiza uralensis Fisch. ex DC.), Chloriti Lapis, and Gypsum Fibrosum (Their ratio is 9.45 : 47.25: 94.5 : 31.5: 15.75 : 31.5: 15.75 : 23.62). A large number of clinical studies have proved that JZOL has a good antiviral effect and can treat lung injury, pneumonia, and bronchitis caused by a variety of viral infections. AIM OF THE STUDY: Influenza infection frequently exhibit dysregulation of gut microbiota and host metabolomes, but the mechanism of JZOL is still unclear and needs to be further explored. Here, after influenza virus infection induced lung injury, the regulation roles of JZOL in metabolic and gut microbiota balances are investigated to comprehensively elucidate its therapeutic mechanism. MATERIALS AND METHODS: A mouse model of lung injury was replicated via intranasal instillation of influenza A (H1N1). The efficacy of JZOL was evaluated by pathological sections, lung index, the levels of TNF-α and IFN-γ, and viral load in lung tissue. Its modulation of endogenous metabolites and gut microbiota was assessed using plasma metabolomic technique and 16S rRNA high-throughput sequencing technique. RESULTS: JZOL not only significantly relieved lung inflammation and edema in influenza mice, but also alleviated the disturbance of endogenous metabolites and the imbalance of gut microbiota mainly by regulating glycerophospholipid and fatty acid metabolism and Lactobacillus. The anti-influenza effects of JZOL were gut microbiota dependent, as demonstrated by antibiotic treatment. The altered metabolites were significantly correlated with Lactobacillus and pharmacodynamic indicators, further confirming the reliability of these results. CONCLUSIONS: JZOL attenuates H1N1 influenza infection induced lung injury by regulating lipid metabolism via the modulation of Lactobacillus. The results support the clinical application of JZOL, and are useful to further understand the mechanism of TCM in the treatment of influenza.

Quality evaluation of ginkgo biloba leaves based on non-targeted metabolomics and representative ingredient quantification.

Ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) combined with multivariate statistical analysis was applied to the study of plant metabolomics to reveal the factors affecting the content of ginkgo leaf compounds. As a follow-up analysis, the terpene lactones and ginkgolic acids were quantified simultaneously using ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLC-QqQ-MS/MS), and subsequently total flavonol glycosides were quantified by high-performance liquid chromatography (HPLC). The results revealed that a total of 52 compounds were potentially identified by establishing a database, and 10 compounds were verified by reference standards; terpene lactones, ginkgolic acids, and flavonoids were the differential compounds; and ginkgolide A was identified as an important indicator compound for tree age. In addition, quantitative analysis showed that the contents of total flavonol glycosides and terpene lactones were highest during April and August in young ginkgo leaves, and differed based on origin. In summary, numerous compounds were rapidly detected by liquid chromatography coupled with MS, the ginkgo leaf samples were compared, and the differential metabolites were screened out. The content changing rules of the target compounds in ginkgo leaves from different regions with different tree ages and harvesting periods were clarified.

[Changes in physicochemical property and microbial community of Pseudostellaria heterophylla soil at diffe-rent fallow ages]. / 太子参不同休耕年限土壤理化特征和微生物群落变化.

Pseudostellaria heterophylla is authentic traditional Chinese herbal medicine in Fujian Province. P. hete-rophylla suffers from serious consecutive monoculture problems. Fallow can alleviate such problems, but the mecha-nism is still unclear. In this study, high-throughput sequencing was performed to analyze the changes in soil microbial community structure and diversity in the P. heterophylla soil at different fallow ages as well as their relationships with soil physicochemical properties and phenolic acids. The results showed that fungal community diversity decreased but bacterial community diversity increased in fallow soils compared with the control soil of P. heterophy-lla. For bacterial communities, the relative abundance of Acidobacteria increased, while that of Proteobacteria and Actinobacteria decreased in fallow soils. For fungal communities, the relative abundance of dominant phyla had no significant difference between fallow and control soils. Soil acidity and organic matter content showed a trend of weakening and decreasing, respectively, with the increases of fallow years. In addition, with the increases of fallow years, the content of phenolic acids in soil, including benzoic acid and salicylic acid, showed significant decrease, while some other phenolic acids such as p-coumaric acid were accumulated obviously. Taken together, fallow could efficiently ameliorate the structure of soil microbial community and soil properties of P. heterophylla, and thus alleviate the effects of continuous cropping.

Exogenous Arginine Treatment Maintains the Appearance and Nutraceutical Properties of Hard- and Soft-Seed Pomegranates in Cold Storage.

Arginine is a natural preservative; however, its effects on the storage of different cultivars of pomegranates have not been investigated extensively. Therefore, the fruit quality of soft-seed Tunisia and hard-seed Yudazi pomegranates was investigated after treatment with arginine at four concentrations during cold storage for 80 days. Pomegranates treated with 1.0 mM arginine exhibited a relatively lower loss of vitamin C, soluble solid, total phenol, and anthocyanin contents in arils, together with a better fruit appearance. Combined with principal component analysis (PCA), the storage life of fruits treated with 1.0 mM arginine showed a higher correlation with antioxidant enzyme activity (e.g., superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT)) during the first 40 days of cold storage, whereas after 40 days of cold storage, storage life was more dependent on the integrity of the cell membrane affected by malondialdehyde (MDA) content, electrolyte leakage (EL), and hydrogen peroxide (H2O2) accumulation. Arginine treatment contributed significantly to the appearance and inner quality of the hard-seed pomegranate cv. Yudazi fruit during cold storage compared to those of soft-seed Tunisia. Taken together, arginine application combined with cold storage enhanced the nutraceutical properties and marketability of pomegranate fruits.

Metabolic characteristics and pharmacokinetic differences of Qiwei Tongbi oral liquid in rheumatoid arthritis rats.

Qiwei Tongbi oral liquid (QWTB), a classical traditional Chinese medicine formula, is widely used to treat arthritis-related diseases in clinical practice. Currently, in vivo metabolic characteristics and pharmacokinetic studies are lacking. This study analyzed the prototype components of QWTB absorbed in the blood and their metabolic transformation process after intragastric administration and compared the differences in pharmacokinetic properties between healthy and rheumatoid arthritis model rats. In sum, 17 prototype components and 21 related metabolites were identified in the plasma and urine of the treated rats. Metabolites were derived from sinomenine and magnoflorine. Through systematic methodology verification, an accurate and stable detection method for sinomenine and magnoflorine in plasma samples was established and applied to pharmacokinetic research of QWTB. At the three dose levels, the AUC0-∞ (area under the curve) of the two components showed a good positive correlation with the dose (R2 > 0.9). Compared with healthy rats, the Tmax , t1/2z , and AUC of sinomenine were markedly increased, and Cmax was decreased in rheumatoid arthritis model rats, indicating that the rate of absorption and elimination rate decreased, but the body exposure increased. However, there were no significant differences in the pharmacokinetic parameters of magnoflorine under healthy and pathological conditions. In summary, the main active ingredients of QWTB are sinomenine and magnoflorine, which exhibit linear kinetic characteristics within a set dose range, and the rheumatoid arthritis pathological state is more conducive to the absorption and efficacy of sinomenine. The results of this study demonstrate the rationality of the clinical application of the QWTB.

Metabolomics analysis of the therapeutic effects of Qiwei Tongbi oral liquid on rheumatoid arthritis in rats.

Qiwei Tongbi oral liquid (QWTB), a classical traditional Chinese medicine (TCM) formula, has a good therapeutic effect on rheumatoid arthritis (RA) and is widely used in China. To comprehensively elucidate the therapeutic mechanism of QWTB in the treatment of RA, the effects of QWTB on biomarkers and metabolic pathways in a rat model of kidney deficiency arthritis were investigated in this study. The effects of QWTB on pharmacodynamic indicators, including paw swelling, arthritis score; interleukin-1ß, interleukin-6, interleukin-17 F, tumor necrosis factor-α, tartrate-resistant acid phosphatase 5b, bone alkaline phosphatase, bone-specific alkaline phosphatase, bone glaprotein, urea, and creatinine levels; and histopathology, suggested that QWTB significantly improved renal function, inhibited the inflammatory response, and reduced bone loss. In total, 39 differential metabolites were screened by comparing the endogenous components between blank and model rat plasma, among which 16 metabolites were altered by QWTB. The metabolism pathway analysis revealed that α-linolenic acid metabolism, phenylalanine metabolism, sphingolipid metabolism, histidine metabolism and glycerophospholipid metabolism were greatly disturbed. Thus, the biomarkers investigated included (1) α-linolenic acid, (2) hippuric acid, (3) phosphatidylethanolamine (15:0/22:2(13Z,16Z)), (4) phenylpyruvic acid, (5) sphinganine, and (6) urocanic acid. QWTB affected three abnormal biomarkers: (3), (4), and (6). Phenylphruvic acid, sphinganine and urocanic acid were significantly associated with pharmacodynamic indicators, as shown by Pearson correlation analysis. These results indicated that RA-related biomarkers had certain reliability and biological significance. In summary, QWTB regulated the metabolic disorders in rats with RA. Its therapeutic mechanism may involve the regulation of phenylalanine metabolism, histidine metabolism, and glycerophospholipid metabolism. The results of this study are useful for understanding the therapeutic mechanisms of TCM.

N-Acyl-Homoserine Lactone Quorum Sensing Switch from Acidogenesis to Solventogenesis during the Fermentation Process in Serratia marcescens MG1.

N-acyl-homoserine lactone quorum sensing (AHL-QS) has been shown to regulate many physiological behaviors in Serratia marcescens MG1. In the current study, the effects of AHL-QS on the biosynthesis of acid and neutral products by S. marcescens MG1 and its isogenic ∆swrI with or without supplementing exogenous N-hexanoyl-L-homoserine lactone (C6-HSL) were systematically investigated. The results showed that swrI disruption resulted in rapid pH drops from 7.0 to 4.8, which could be restored to wild type by supplementing C6-HSL. Furthermore, fermentation product analysis indicated that ∆swrI could lead to obvious accumulation for acidogenesis products such as lactic acid and succinic acid, especially excess acetic acid (2.27 g/l) produced at the early stage of fermentation, whereas solventogenesis products by ∆swrI appeared to noticeably decrease by an approximate 30% for acetoin during 32-48 h and by an approximate 20% for 2,3-butanediol during 24-40 h, when compared to those by wild type. Interestingly, the excess acetic acid produced could be removed in an AHL-QS-independent manner. Subsequently, quantitative real-time PCR was used to determine the mRNA expression levels of genes responsible for acidogenesis and solventogenesis and showed consistent results with those of product synthesis. Finally, by close examination of promoter regions of the analyzed genes, four putative luxI box-like motifs were found upstream of genes encoding acetyl-CoA synthase, lactate dehydrogenase, α-acetolactate decarboxylase, and Lys-like regulator. The information from this study provides a novel insight into the roles played by AHL-QS in switching from acidogenesis to solventogenesis in S. marcescens MG1.