Ginseng fermentation solution affects the gut microbiota in zebrafish with alcoholic liver disease via PI3K/Akt pathway.

BACKGROUND: Ginsenosides have received increased amounts of attention due to their ability to modulate the intestinal flora, which may subsequently alleviate alcoholic liver disease (ALD). The effects of ginseng fermentation solution (GFS) on the gut microbiota and metabolism in ALD patients have not been explored. PURPOSE: This research aimed to explore the regulatory effect of GFS on ALD both in vitro and in vivo. METHOD: This study assessed the anti-ALD efficacy of GFS using an LO2 cell model and a zebrafish model. Untargeted metabolomics was used for differentially abundant metabolite analysis, and high-throughput 16S rRNA sequencing was used to examine the effect of GFS on ALD. RESULTS: The LO2 cell line experiments demonstrated that GFS effectively mitigated alcohol-induced oxidative stress and reduced apoptosis by upregulating PI3K and Bcl-2 expression and decreasing the levels of malondialdehyde, total cholesterol, and triglycerides. In zebrafish, GFS improved morphological and physiological parameters and diminished oxidative stress-induced ALD. Meanwhile, the results from Western blotting indicated that GFS enhanced the expression of PI3K, Akt, and Bcl-2 proteins while reducing Bax protein expression, thereby ameliorating the ALD model in zebrafish. Metabolomics data revealed significant changes in a total of 46 potential biomarkers. Among them, metabolites such as prostaglandin F2 alpha belong to arachidonic acid metabolism. In addition, GFS also partly reversed the imbalance of gut microbiota composition caused by alcohol. At the genus level, alcohol consumption elevated the presence of Flectobacillus, Curvibacter, among others, and diminished Elizabethkingia within the intestinal microbes of zebrafish. Conversely, GFS reversed these effects, notably enhancing the abundance of Proteobacteria and Archaea. Correlation analyses further indicated a significant negative correlation between prostaglandin F2 alpha, 11,14,15-THETA, Taurocholic acid and Curvibacter. CONCLUSION: This study highlights a novel mechanism by which GFS modulates anti-ALD activity through the PI3K/Akt signalling pathway by influencing the intestinal flora-metabolite axis. These results indicate the potential of GFS as a functional food for ALD treatment via modulation of the gut flora.

Identifying the active compounds and mechanism of action of TongFu XieXia Decoction for treating intestinal obstruction using network pharmacology combined with ultra-high performance liquid chromatography-quadrupole-orbitrap mass spectrometry.

RATIONALE: TongFu XieXia Decoction (TFXXD), a formulation rooted in traditional Chinese medicine and optimized through clinical practice, serves as an advanced version of the classic Da Cheng Qi decoction used for treating intestinal obstruction (IO), demonstrating significant therapeutic efficacy. However, due to the intricate nature of herbal compositions, the principal constituents and potential mechanisms of TFXXD have yet to be clarified. Accordingly, this study seeks to identify the active compounds and molecular targets of TFXXD, as well as to elucidate its anti-IO mechanisms. METHODS: Qualitative identification of the principal constituents of TFXXD was accomplished using ultra-high preformance liquid chromatography-quadrupole-orbitrap mass spectrometry (UPLC-Q-Orbitrap-MS/MS) analysis. PharmMapper facilitated the prediction of potential molecular targets, whereas protein-protein interaction analysis was conducted using STRING 11.0. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed using the Metascape database. A "compounds-target-pathway" network was meticulously constructed within Cytoscape 3.8.2. Finally, molecular docking studies were performed to investigate the interactions between the core target and the crucial compound. RESULTS: UPLC-Q-Orbitrap-MS/MS analysis identified 65 components with high precision and sensitivity. Furthermore, 64 potential targets were identified as integral to TFXXD bioactivity in IO treatment. Gene Ontology enrichment analysis revealed 995 distinct biological functions, while the Kyoto Encyclopedia of Genes and Genomes enrichment analysis identified 143 intricate signaling pathways. CONCLUSION: Molecular docking studies substantiated the substantial affinity between the TFXXD bioactive constituents and their corresponding targets in the context of IO. TFXXD exerts its therapeutic efficacy in IO through a multifaceted interplay between multiple compounds, targets, and pathways. The integration of network pharmacology with UPLC-Q-Orbitrap-MS/MS has emerged as a promising strategy to unravel the intricate web of molecular interactions underlying herbal medicine. However, it is imperative to emphasize the necessity for further in vivo and in vitro experiments.

Effects of ginseng on short-chain fatty acids and intestinal microbiota in rats with spleen-qi deficiency based on gas chromatography-mass spectrometry and 16s rRNA technology.

RATIONALE: Spleen-qi deficiency syndrome, a common weakness syndrome in traditional Chinese medicine, results from insufficient spleen-qi levels. For centuries, ginseng has been relied upon as a traditional Chinese medicine to treat spleen-qi deficiency syndrome. Until now, the mechanism feature of ginseng in treating temper deficiency through intestinal bacteria and short-chain fatty acid (SCFA) metabolites has not been fully elucidated. METHODS: This study established a rat model of spleen-qi deficiency via multi-factor compound modeling that involved fatigue injury and a controlled diet. The content of SCFAs between different treatment groups was determined by gas chromatography-mass spectrometry. And the 16s rRNA sequencing technology was applied to reveal the effects of ginseng on the intestinal microecological environment of the rats. RESULTS: It was found that the ginseng treatment group exhibited the most remarkable regulatory effect on propionic acid, surpassing all other administration groups. Ginseng increased the relative abundance of beneficial bacteria and decreased that of harmful bacteria at the genus level in rats with spleen-qi deficiency syndrome. And propionic acid is significantly positively correlated with Lactobacillus level and significantly negatively correlated with uncultured_bacterium_f_Muribaculaceae (p < 0.05). n-Butyric acid is negatively correlated with the Faecalibaculum level (p < 0.01). n-Valeric acid is significantly negatively correlated with the Romboutsia level (p < 0.01). CONCLUSION: The mechanism of ginseng treatment for spleen-qi deficiency is elucidated from the perspective of gut microbiota and its metabolite SCFAs. It provides a new way for further development and utilization of ginseng and a theoretical basis.

Immunomodulatory effect of Liangyi paste on the gut microbiota of mice.

Liangyi paste (LY) is a traditional Chinese medicine made from a mixture of Ginseng and Rehmanniae radix praeparata. The present study aimed to investigate the effects of LY on gut microbiota diversity in immunocompromised mice. The chemical composition of LY extract was analyzed using UPLC-Q-Orbitrap-MS/MS, and the differences in the structure and diversity of the intestinal microbiota of LY extract were examined using 16S rRNA. In this study, identified and analyzed 66 compounds from the LY. These compounds included 11 iridoids, 6 oligosaccharides, 21 protopanaxtriols, 23 protopanaxadiols, 2 OLE, 1 Ionone and 2 phenylethanoside, using advanced UPLC-Q-Orbitrap-MS/MS technology. Through the use of 16S rRNA analysis, the study found that LY significantly increased the relative abundance of the Firmicutes phylum in immunocompromised mice, while decreasing the abundance of the Proteobacteria and Actinobacteria phyla. At the genus level, LY significantly increased the relative abundance of beneficial bacteria such as Clostridium_sensu_stricto_l, Lactobacillus, and Limosilactobacillus in immunocompromised mice. Conversely, the paste extract decreased the relative abundance of harmful bacteria such as Enterococcus and Escherichia Shigella in immunocompromised mice. These findings highlight the potential of LY to serve as a natural dietary supplement for enhancing gut microbiota diversity and promoting gut health. The identification of numerous compounds within the paste extract demonstrates its complexity and potential as a source for further research and development. Additionally, the LY extract exerted a significant influence on both nucleotide and amino acid metabolism. To sum up, the findings suggest that the LY extract has the potential to modulate the structure and diversity of gut microbiota, as well as promote metabolic balance in immunocompromised mice.

Urine and serum metabolomics study of wild ginseng in rats with spleen-qi deficiency using rapid resolution liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry.

Patients with a spleen-qi deficiency often exhibit dysfunction in the metabolic system. Metabolites are considered the most direct reflection of individual physiological and pathological conditions and represent attractive candidates to provide deep insights into disease phenotypes. This study examines the potential therapeutic mechanism of wild ginseng on spleen-qi deficiency through the analysis of serum and urine metabolomics using rapid-resolution liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry. The reasons for the superiority of wild ginseng treatment over cultivated ginseng were also analyzed in depth. After wild ginseng intervention, anandamide, urobilinogen, aldosterone, and testosterone glucuronide were significantly reduced in serum. Meanwhile, argininosuccinic acid, L-cysteine, and seven other metabolites were significantly elevated in serum. Nine metabolites, including L-acetylcarnitine, and citrulline were elevated in the urine, and trimethylamine N-oxide, adrenic acid, and 10 other metabolites were reduced. Arginine biosynthesis, pantothenate and CoA biosynthesis, thiamin metabolism, taurine, and tryptophan metabolism pathways were mainly improved. Further analysis was conducted on the relationship between Lactobacillus and Akkermansia bacteria with metabolites, and it was found that they are mainly related to amino acid metabolites. This study provides strong theoretical support and direction for further explanation of the immune mechanism of wild ginseng and lays the foundation for future studies.

Metabolomics and network pharmacological analysis to explore the mechanism of ginseng treatment for spleen-qi deficiency.

Ginseng is the main Chinese herbal medicine for tonifying Qi and invigorating the spleen. It has been used to treat spleen-qi deficiency with good protective effects for thousands of years, however, its biological mechanism has not been fully elucidated. This study aims to explore the mechanism of ginseng in the treatment of spleen-qi deficiency by using a comprehensive method combining metabolomics and network pharmacological analysis. Gas chromatography-mass spectroscopy was applied for investigating the changes in urine metabolites in spleen-qi deficiency rats and after treatment with ginseng. Metabolomics and network pharmacology analysis were applied to screen potential biomarkers and therapeutic targets of ginseng in the treatment of spleen-qi deficiency, respectively. Molecular docking was employed to further evaluate the docking mode of potential biomarkers and therapeutic target proteins. The results of metabolomics showed that the therapeutic effects of ginseng are mainly related to its regulation of three metabolic pathways. The molecular structure of potential biomarkers and common proteins was further analyzed by molecular docking to verify its effectiveness. Ginseng has good pharmacological effects by controlling key targets of related metabolic pathways, signal pathways, and potential biomarkers.

Treatment of Obese Zebrafish with Saringosterol Acetate through AMP Activated Protein Kinase Pathway.

OBJECT: Edible Brown Seaweed Sargassum fusiforme (Harvey) Setchell, 1931 abbreviated as Sargassum fusiforme was used for folk medical therapy in East Asia countries over five hundred years. Saringosterol acetate (SA) was isolated from S. fusiforme in our previous study and indicated various effects. However, anti-obesity activity of SA and its mechanism still unknown. METHOD: The inhibitory effect of SA, isolated from S. fusiforme, on adipogenesis in 3T3-L1 adipocytes was investigated in vitro and in zebrafish model. Cell toxicity, differentiation, signaling pathway, and lipid accumulation of SA treated 3T3-L1 adipocytes were determined. The body weight and triglyceride content of diet-induced obese (DIO) adult male zebrafish were measured from 12 to 17 weeks after fertilization. RESULT: SA attenuated the differentiation of cells and reduced lipid accumulation, and triglyceride content in the 3T3-L1 adipocytes. During the differentiation of adipocytes, SA suppressed fat accumulation and decreased the expression of signal factors responsible for adipogenesis. In SA-treated adipocytes, while fatty acid synthetase was downregulated, AMP-activated protein kinase (AMPK) was upregulated. Furthermore, SA suppressed body weight and triglyceride content in DIO zebrafish. CONCLUSION: SA is a potential therapeutic agent in the management of metabolic disorders, such as obesity.

Intervention effects of ginseng on spleen-qi deficiency in rats revealed by GC-MS-based metabonomic approach.

Panax ginseng C. A. Mey. (Ginseng) is a famous Chinese medicine with tonifying middle and replenishing qi effects and has been applied for the treatment of spleen-qi deficiency for many years. However, its potential therapeutic mechanisms have not been thoroughly studied. In this study, the metabolomic technique was applied to explore the therapeutic effect of ginseng on the spleen-qi deficiency. A rat model of spleen-qi deficiency was generated via the fatigue swimming method. After 3 weeks of treatment with ginseng, the entire metabolic changes in rat serum were profiled by gas chromatography-mass spectroscopy (GC-MS). The metabolic profiles in serum taurine and hypotaurine metabolism significantly differed among groups, in which a total of 17 metabolites were identified. Ginseng reversed the metabolic changes in the difference involving some metabolic pathways. Among them, beta-alanine metabolism, taurine and hypotaurine metabolism, and the pentose phosphate pathway are the key metabolic pathways. The therapeutic effects of ginseng on spleen-qi deficiency rats could be achieved by regulating multiple metabolic pathways, metabolites can be used as potential biomarkers for the diagnosis and monitoring of spleen-qi deficiency.

Effects of Scutellaria baicalensis Georgi. on intestinal flora in rats with spleen deficiency and damp-heat.

The effects of Scutellaria baicalensis Georgi. (S. baicalensis Georgi.) on the diversity of intestinal flora in rats with spleen deficiency and damp-heat was explored in the present study. 51 compounds in S. baicalensis Georgi. extract, including 37 flavonoids, 9 dihydroflavone, and 5 flavanols, were identified by ultra-high performance liquid chromatography-Q-Orbitrap-mass spectrometry(UPLC-Q-Orbitrap-MS/MS). Ethanol extract from Scutellariae Radix and fresh feces from rats with spleen deficiency and damp-heat were incubated in vitro for 48 h. At the phylum level, the ethanol extract noticeably increased the relative abundance of Firmicutes in the feces and effectively reduced those of Proteobacteria and Actinobacteria. At the genus level, the extract increased the relative abundance of the Lactobacillus and Bifidobacterium and reduced those of pathogenic bacteria, including Clostridium, Escherichia, Enterococcus, and Streptococcus. The results suggest that S. baicalensis Georgi. can regulate the structure and diversity of intestinal flora in rats with spleen deficiency and damp-heat and balance the body's metabolism.

In Vitro Transformation of Protopanaxadiol Saponins in Human Intestinal Flora and Its Effect on Intestinal Flora.

Protopanaxadiol (PPD)-type ginsenosides are the main ginsenosides in ginseng (Panax ginseng C. A. Meyer) with potential therapeutic effects on diseases related to intestinal flora imbalance. This study aimed to investigate the in vitro metabolism of protopanaxadiol ginsenosides in human intestinal flora and their effect on the flora. Rapid resolution liquid chromatography coupled with quadruple-time-of-flight mass spectrometry (RRLC-Q-TOF MS) was utilised for the transformation of ginsenoside constituents for sample identification. Using 16S rDNA gene sequencing technique, the effect of PPD-type ginsenosides on gut microflora was analysed based on the indices of microflora diversity and gut microflora. The sample was transformed for 6 h, and the metabolites were ginsenoside Rb1, Rc, Rb2, Rb3, CO, Gyp-IX, Gyp-XVII, CMc-1, F2, Rg3, CK, Rh2, and PPD. The metabolites were CK, Rh2, and PPD when the samples were transformed for 60 h. The intestinal microflora were subjected to high-throughput sequencing using the Illumina MiSeq 2500 sequencing platform. In comparison with the faecal sample from the blank group, the protopanaxadiol saponin group significantly increased the relative abundance of Firmicutes and significantly decreased Bacteroidetes and Proteobacteria at the phylum level, whereas it significantly increased the relative abundance of Prevotella_9, Faecalibacterium, and Dialister and significantly decreased Escherichia-Shigella, Dorea, and Lachnoclostridium at the genus level. This study provides a basis for the determination of the pharmacodynamic material basis and pharmacodynamic targets of PPD-type ginsenosides based on the intestinal flora.

In Vitro Effects of Ginseng and the Seed of Zizyphus jujuba var. spinosa on Gut Microbiota of Rats with Spleen Deficiency.

Ginseng and the seed of Zizyphus jujuba var. spinosa, which are traditional Chinese medicinal materials, were often used in ancient Chinese recipes as a pair of medicines. They can replenish the primordial qi and tonify the spleen. This study investigated the effects of ginseng and the seed of Zizyphus jujuba var. spinosa (GS) extract on gut microbiota diversity in rats with spleen deficiency syndrome (SDS). A total of 52 compounds (including 16 flavonoids, 35 saponins, and 1 alkaloid) were identified and analyzed from the GS extract by UPLC-Q-Orbitrap-MS/MS. The GS extract significantly increased the relative abundance of Firmicutes and Bacteroidetes in rats with SDS but decreased that of Proteobacteria and Actinobacteria. At the genus level, the GS extract significantly increased the relative abundance of Lactobacillus and Bifidobacterium in rats with SDS but decreased that of Streptococcus, Escherichia-Shigella, Veillonella, and Enterococcus. In addition, the GS extract influenced glucose and amino acid metabolism. In summary, the results showed that the GS extract changed the structure and diversity of gut microbiota in rats with SDS and balanced the metabolic process.