Epimedium (EM), also known as barrenwort, is a traditional medicinal plant rich in isopentenyl flavonols, which have beneficial biological activities and can improve human and animal health, but its mechanism is still unclear. In this study, ultra-high-performance liquid chromatography/quadrupole-time-of-flight-mass spectrometry (UHPLC-Q-TOF/MS) and ultra-high-performance liquid chromatography triple-quadrupole mass spectrometry (UHPLC-QqQ-MS/MS) were used to analyse the main components of EM, and isopentenyl flavonols such as Epimedin A, B, and C as well as Icariin were the major components of EM. Meanwhile, broilers were selected as model animals to illuminate the mechanism of Epimedium isopentenyl flavonols (EMIE) on gut health. The results showed that supplementation with 200 mg/kg EM improved the immune response, increased cecum short-chain fatty acids (SCFAs) and lactate concentrations, and improved nutrient digestibility in broilers. In addition, 16S rRNA sequencing showed that EMIE altered the composition of cecal microbiome, increasing the relative abundance of beneficial bacteria (Candidatus Soleaferrea and Lachbospiraceae NC2004 group and Butyricioccus) and reducing that of harmful bacteria (UBA1819, Negativibacillus, and Eisenbergiella). Metabolomic analysis identified 48 differential metabolites, of which Erosnin and Tyrosyl-Tryptophan were identified as core biomarkers. Erosnin and tyrosyl-tryptophan are potential biomarkers to evaluate the effects of EMIE. This shows that EMIE may regulate the cecum microbiota through Butyricicoccus, with changes in the relative abundance of the genera Eisenbergiella and Un. Peptostreptococcaceae affecting the serum metabolite levels of the host. EMIE is an excellent health product, and dietary isopentenyl flavonols, as bioactive components, can improve health by altering the microbiota structure and the plasma metabolite profiles. This study provides the scientific basis for the future application of EM in diets.

Background: Caffeic acid (CA) and its derivative, chlorogenic acid (CGA), have shown promise in preventing and alleviating fatty liver disease. CA, compared to CGA, has much lower production costs and higher bioavailability, making it a potentially superior feed additive. However, the efficacy, mechanistic differences, and comparative impacts of CA and CGA on fatty liver disease in laying hens remain unclear. This study aimed to evaluate and compare the effects of CA and CGA on production performance, egg quality, and fatty liver disease in laying hens. Results: A total of 1,440 61-week-old Hyline Brown laying hens were randomly divided into 8 groups and fed diets supplemented with basal diet, 25, 50, 100 and 200 mg/kg of CA, and 100, 200 and 400 mg/kg of CGA (CON, CA25, CA50, CA100, CA200, CGA100, CGA200 and CGA400, respectively) for 12 weeks. Both CA and CGA improved production performance and egg quality, while reducing markers of hepatic damage and lipid accumulation. CA and CGA significantly decreased TG, TC, and LDL-C levels and increased T-SOD activity. Transcriptomic and proteomic analyses revealed that CA and CGA reduced hepatic lipid accumulation through downregulation of lipid biosynthesis-related genes (ACLY, ACACA, FASN, and SCD1) and enhanced lipid transport and oxidation genes (FABPs, CD36, CPT1A, ACOX1, and SCP2). Of note, low-dose CA25 exhibited equivalent efficacy to the higher dose CGA100 group in alleviating fatty liver conditions. Mechanistically, CA and CGA alleviated lipid accumulation via activation of the ADPN-AMPK-PPARα signaling pathway. Conclusions: This study demonstrates that dietary CA and CGA effectively improve laying performance, egg quality, and hepatic lipid metabolism in laying hens, with CA potentially being more economical and efficient. Transcriptomic and proteomic evidence highlight shared mechanisms between CA25 and CGA100. These findings provide a foundation for CA and CGA as therapeutic agents for fatty liver disease and related metabolic diseases in hens, and also offer insights into the targeted modification of CGA (including the isomer of CGA) into CA, thereby providing novel strategies for the efficient utilization of CGA. Graphical

Gut microbial β-glucuronidase (GUS) plays a pivotal role at the microbiota—host interface by hydrolyzing glucuronide conjugates, thereby influencing xenobiotic metabolism, enterohepatic circulation, and systemic homeostasis. Dysregulated GUS activity has been increasingly linked to adverse health outcomes, including drug-induced toxicity, inflammation, and cancer. However, current literature often overlooks the enzyme’s dual role in maintaining physiological balance and promoting disease progression, as well as the multidimensional ways in which natural products interact with GUS. This work reviews recent advances in GUS research, emphasizing its structural diversity, functional complexity, and regulatory impact on host health. It also highlights the potential of natural products as precision modulators of GUS activity, capable of direct enzyme inhibition or indirect modulation through reshaping the gut microbiota. These mechanisms collectively influence drug efficacy, toxicity, and the systemic availability of endogenous metabolites. By integrating structural, pharmacological, and microbiological perspectives, this work provides a theoretical foundation for the development of microbiota-targeted therapies centered on GUS. Such approaches may support the rational design of natural product-derived inhibitors and promote their application in disease models, ultimately advancing personalized therapeutic strategies.

Dietary flavonoids are regarded as potent inhibitors of gut bacterial β-glucuronidase (β-GUS), although the precise structure-activity relationship between dietary flavonoids and β-GUS remains elusive. In this study, 24 dietary flavonoids of 6 common different structural types were selected as model molecules to investigate this relationship through enzymatic inhibition kinetics and molecular docking simulations. The findings revealed that delphinidin (1) and baohuoside I (16) exhibited robust inhibitory activities, while the formation of ionic bond and hydrogen bonds was found to play crucial roles in these interactions. Moreover, the core structure, position, and quantity of hydroxyl groups; type and number of glycosyls, and the presence of isopentenyl group all played pivotal roles in influencing inhibitory activities against β-GUS derived from Escherichia coli. These results offer valuable insights into the relationship between flavonoid structure and enzyme inhibition, which could prove invaluable for the development of novel drugs and flavonoid dietary supplements.

This study aimed to explore the effects of Galega orientalis Lam. Flavonoids Extract (GOLFE) on broiler's meat and the gut microbiota while predicting the plant's mechanism of action through network pharmacology analysis. GOLFE was first analyzed using UHPLC-Q Exactive HRMS to identify the bioactive compounds responsible for its potential effects. Subsequently, a total of three hundred and sixty, one-day-old Cobb broiler chicks, divided into four groups, including a negative control CON fed a basal diet, two treatment groups with 200 mg and 400 mg GOLFE, and a positive control CT with 100 mg Chlortetracycline, were used for a 42-day trial. The results revealed that GOLFE increased the breast meat weight, redness (a*), catalase level, and superoxide dismutase activity, while reducing the malondialdehyde content (P < 0.05). The extract also altered the gut microbiota content by reducing the level of Clostridia UCG-014 and increasing the presence of butyrate-producing bacteria, notably Butyricicoccus and Flavonifractor, in the gut microbiota (P < 0.05). Furthermore, through a network pharmacology analysis, the identified bioactive components, namely, Quercetin 3-O-rutinoside-(1-2)-O-rhamnoside, Clitorin, Rutin, Isoquercitrin, Kaempferol-3-O-rutinoside, Narcissin, and Quercitrin, helped predict that GOLFE could enhance the meat antioxidant capacity by modulating broiler's gut microbiota, possibly through the MAPK signaling pathway. These findings potentially position GOLFE as a sustainable additive for broiler feed.