Cyclophosphamide (CTX) is a common anticancer drug that can cause immunosuppression and disrupt the intestinal microbiota when taken long-term or at high doses. Lonicerae japonicae Flos polysaccharides (LJFP), identified as the principal bioactive constituents of L. japonicae Flos, have emerged as promising immunomodulatory agents. Herein, we explored the molecular mechanisms through which LJFP alleviates CTX-induced immunosuppression in mice. As determined via gas chromatography, LJFP contains arabinose, rhamnose, ribose, xylose, mannose, fructose, galactose, and glucose. LJFP mitigated the decline in body weight and immune organ indexes in CTX-induced mice, while promoting cytokine production. Moreover, LJFP stimulated immunity by reshaping the gut microbiota of mice, where norank_f_Muribaculaceae and Alistipes played key roles. LJFP also increased short-chain fatty acid levels in the mouse colon and feces. Network pharmacology analyses identified 31 core targets and relevant signaling pathways implicated in the treatment of immunosuppression. Molecular docking indicated strong binding of the eight monosaccharide components to key targets, epidermal growth factor receptor, heat shock protein 90 alpha family class member 1, and interleukin 2. Notably, 15 of these targets are specific to the gut, further suggesting that the mechanism through which LJFP ameliorates immunosuppression is intricately linked to the gut microbiota. The current findings highlight the potential of LJFP as a functional food or alternative therapeutic against immunosuppression. (c) 2027 The authors. Publishing services by Elsevier B.V. on behalf of KeAi Communications Co., Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Lonicerae japonicae flos (LJ), derived from Lonicera japonica Thunb., is a classic medicinal and edible plant traditionally used to clear heat, detoxify, and disperse wind-heat. It is widely consumed as a health tea or food additive and played a significant role during the COVID-19 pandemic. Its closely related species, Lonicerae flos (LFS), derived from L. macranthoides, L. hypoglauca, L. confusa, and L. fulvotomentosa, share similar functions. Distinguishing between these two is crucial to minimizing medication-related risks; however, studies on their differences in inorganic elements are currently insufficient. This study investigated the content differences in ten inorganic elements in 97 samples using inductively coupled plasma mass spectrometry and inductively coupled plasma atomic emission spectrometry. LJ exhibited significantly higher iron (Fe) content and considerably lower manganese (Mn) content than LFS. Fe, Mn and Na were identified as key discriminatory elements through orthogonal partial least squares-discriminant analysis, and cluster analysis based on the Fe/Mn ratio also achieved clear separation between the two groups. Network pharmacology and pathway enrichment analyses were subsequently conducted to explore the potential biological relevance of Fe- and Mn-associated differences. Additionally, molecular docking provided exploratory computational assessments of theoretical interactions between Fe/Mn and selected target proteins under simplified conditions. It is important to note that these computational results should not be interpreted as direct evidence of biological activity, since no in vitro or in vivo validation was performed. Overall, this study provides elemental profiling data to differentiate between LJ and LFS and highlights the potential value of the Fe/Mn ratio as a complementary chemical indicator for the authentication and quality evaluation of Lonicera medicinal materials.

Cerasus humilis (Bge.) Sok. fruits have high proanthocyanidin (PA) content. We aimed to identify and perform preliminary functional evaluations of four candidate transport-related genes to investigate their roles in PA accumulation. We analyzed the gene expression patterns, determined the subcellular localization of associated proteins, conducted phylogenetic analyses to examine their evolutionary orthologs, evaluated their roles in PA accumulation through overexpression assays, and performed molecular docking simulations to analyze binding between these candidate transporters and the specific PA monomers catechin and epicatechin. Together, ChGST2 , ChMATE1 , and ChMATE2 expression levels were correlated with PA accumulation; ChGST1 and ChGST2 were localized in the cytoplasm and nucleus; ChMATE1 and ChMATE2 were localized in the vacuolar membrane (tonoplast). Phylogenetic analyses confirmed that ChGST1/2 and ChMATE1/2 are true orthologs of well-characterized plant PA transporters. Furthermore, overexpression of ChGST2 , ChMATE1 , and ChMATE2 enhanced PA accumulation, indicating ChGST2 and ChMATE1/2 play roles in the PA accumulation pathway in C. humilis . Our findings implied GSTs and MATE genes might be important for the nutritional value of berries via enhancing the PA accumulation.

Directed fermentation has emerged as a key strategy for enhancing the sensory quality and health benefits of plant-based foods. However, the mechanisms by which specific microbial strains remodel metabolic networks within complex plant matrices to co-regulate flavor formation and bioactive compound accumulation remain unclear. In this study, Lactiplantibacillus plantarum BUCM-1, a strain with lipid-lowering potential isolated from traditional fermented foods, was employed to establish a directed fermentation model for mung bean beverage. Integrated multi-omics analyses revealed that BUCM-1-dominated fermentation reshaped both the volatile and non-volatile metabolic profiles, accompanied by enhanced in vitro lipid-lowering activity, with lipid reduction improving from 26% to 88%. Off-odor compounds were suppressed, while aroma-active compounds were enriched. At the non-volatile level, biotransformation was mainly centered on amino acid metabolism and the tricarboxylic acid cycle. Notably, the aspartate-ornithine metabolic axis was strongly activated, with aspartate and ornithine increasing by approximately 7-and 30-fold, respectively, suggesting enhanced coupling between nitrogen metabolism and central carbon metabolism during fermentation. Overall, this study demonstrates that fermentation with BUCM-1 enables the synergistic improvement of flavor quality and functional activity in a high-protein plant substrate. These findings provide mechanistic insights and a metabolomics-guided strategy for developing functional plant-based fermented beverages with controllable flavor and targeted health benefits.

Lonicera japonica Thunb. possesses significant potential for applications in beverages and functional foods. Nevertheless, most studies focus on the overall quality of flower buds, with limited comparisons across different developmental stages and distinct floral parts. This study systematically investigated changes in volatile flavor compounds and non-volatile bioactive constituents in whole flowers, calyxes, corollas, and reproductive organs before and after flowering. An integrated approach combining metabolomics profiling, entropy weight analysis, correlation network analysis, and molecular docking was employed to evaluate their potential in functional food development. The level of 3-decyn-2-ol increased markedly after flowering, with an approximately 14-fold increase in reproductive organs. Phenolic acids were highly enriched in the calyx, reaching up to 12-fold higher than in other parts. Flavonoids predominated in the corolla at levels 1.5-3-fold higher than in other tissues. Following flowering, the overall levels of phenolic acids and flavonoids decreased, while total sugars, reducing sugars, and polysaccharides increased by approximately 59%, 98%, and 35%, respectively. These results suggest that open flowers may exhibit enhanced potential for functional food applications. Entropy weight analysis indicated that the calyx contributed most to the integrated evaluation of flavor and functional attributes. Correlation network analysis identified chlorogenic acid, neochlorogenic acid, rutin, luteoloside, loganic acid, and secologanoside as key constituents, which showed potential interactions with inflammation- and immunity-related targets in molecular docking. These findings suggest that although medicinal use decreased after flowering, the edible value of L. japonica may increase, providing a basis for its rational utilization in functional food development.

ETHNOPHARMACOLOGICAL RELEVANCE:Bile acids (BAs) have long been recognized in traditional ethnic medicines as a regulator of emotion and mental states; however, the underlying biological mechanisms by which BAs influence neuropsychological functions remain largely unclear. Multiflorin A (MA), an acetylated flavonoid glycoside and the signature bioactive constituent of Pruni Semen, is believed to ameliorate psychological stress via targeting the bile system. AIM OF THE STUDY:This study aimed to investigate alterations in bile acid metabolism and distribution in SPS-induced PTSD-associated anxiety-like behavioral alterations and the therapeutic effects of MA and ursodeoxycholic acid (UDCA). MATERIALS AND METHODS:SPS-stressed mice exhibiting anxiety-like behaviors were treated with MA. Behavioral tests, histopathology, targeted BAs metabolomics, metagenomics, neurotransmitter profiling, proteomics, and immunofluorescence were performed. UDCA was used as a reference compound to explore the involvement of BAs in MA-mediated neuroprotective effects. RESULTS:SPS exposure induced anxiety-like behavioral deficits, accompanied by dysregulation of systemic BAs homeostasis, characterized by peripheral BAs depletion, central accumulation of hydrophobic BAs, partial blood-brain barrier disruption, and synaptic impairment. MA and UDCA treatment significantly improved behavioral performance, alleviated histopathological damage, and partially restored gut microbiota composition and BAs profiles, including increased levels of isomerized bile acids such as UDCA and alloLCA. These changes were accompanied by restoration of tight junction, PSD-95 expression, and neurotransmitter balance. Proteomics showed partial reversal of SPS-induced synaptic and neurotransmitter dysregulation, consistent with reduced neural hyperexcitability. CONCLUSION:MA may ameliorate PTSD-associated anxiety-like behavioral alterations through modulation of the gut microbiota-bile acid-brain interactions, supporting a role for BAs metabolism in neuropsychiatric regulation.

Constipation is the most common gastrointestinal disease in clinical practice. Although anthraquinone have proven effective for constipation treatment, long-term use is associated with adverse reactions, indicating the need for novel non-anthraquinone drugs. Our previous studies identified multiflorin A from the Pruni coat of Cerasus humilis as a potent laxative agent. However, the comprehensive composition profile of semen coat extracts (PSCE) and their collective mechanisms remain elusive. This study systematically gated the extraction optimization, phytochemical characterization, and therapeutic mechanisms of PSCE. soaking extraction exhibited optimal efficiency for PSCE preparation. UPLC-MS/MS analysis identified flavonoid compounds in PSCE, with multiflorin A, multiflorin B, afzelin, juglalin, and kaempferol constituting predominant constituents. Molecular docking simulations systematically predicted potential protein mediating laxative activity of PSCE. In loperamide-induced constipated mice, medium-dose PSCE (5 significantly enhanced intestinal transit, improved fecal morphology, and increased stool moisture Mechanistic investigations revealed that PSCE (5 g/kg) normalized intestinal dysfunction through differential upregulation of AQP4 in colonic tissues and AQP3 in the small intestine, coupled with activation of c-Kit/SCF signaling pathways in both intestinal regions. These findings position PSCE as a dual-action therapeutic targeting small intestinal and colonic motility regulation, providing a novel therapeutic candidate stipation management.

Obesity and overweight have increasingly posed a serious challenge to public health security. This study systematically evaluated the reversal and regulatory effects of a composite flavonoid component mimicking the composition of adzuki bean flavonoids on high-fat diet (HFD)-induced obesity, related lipid metabolism disorders, and impaired liver function, based on lipid metabolomics and an HFD-induced obese mouse model. The results demonstrated that sustained HFD intake led to significant weight gain, increased adiposity index, dyslipidemia, and altered brown adipose tissue (BAT) cell status in mice, while also exerting adverse effects on hepatic lipid deposition and the lipid metabolic profile associated with liver fibrosis. Intervention with an adzuki bean flavonoid mimic (ABFM) effectively prevented further weight gain and ameliorated abnormal expression of serum lipid and liver function-related indicators. Furthermore, we found that ABFM alleviated HFD-induced liver damage and mitigated the whitening tendency of brown adipose tissue. Lipidomics analysis revealed that ABFM intake significantly improved abnormal hepatic lipid metabolic profiles, notably downregulating the expression levels of diacylglycerol (DG) and phosphatidylglycerol (PG), while markedly ameliorating sphingolipid metabolism disorders and ceramide (Cer) levels, which are highly associated with liver fibrosis. These findings further elucidate the mechanisms by which adzuki bean flavonoid components improve diet-induced obesity and associated liver injury, providing a theoretical basis for exploring safe and effective dietary intervention strategies based on plant flavonoids.

Diabetes mellitus, a chronic metabolic disorder, has become a global health concern due to its increasing prevalence and associated complications. Mulberry (Morus alba), widely used in the food and nutraceutical industries, has shown potential in alleviating metabolic disorders through its protein. This study aimed to analyze the composition of the protease hydrolysate of a new resource species, the protein mulberry, and investigate its activity in ameliorating type 2 diabetes mellitus (T2DM). Proteolytic hydrolysis of protein mulberry leaf proteins increased total amino acid content (TAA) by 11.58%, with significant increases in 14 amino acids. Chromatography and mass spectrometry indicated that the main peptides in hydrolyzed proteins with molecular weight was less than 10 kDa and 802 bioactive peptides were identified. Moreover, this study constructed a mouse model of streptozotocin (STZ)/high-fat diet-induced T2DM.Two peptides showed tolerance to digestive enzymes. Protein mulberry leaf proteolytic hydrolysate (MLPH) effectively improved the blood sugar level and glucose tolerance of diabetic mice; increased the activities of serum C-peptide, liver glycogen, and antioxidant enzymes; reduced the indices of blood lipid and oxidative stress; and improved the morphology of liver and islet cells. Transcriptome analysis showed that the genes upregulated in the MLPH group were primarily involved in the synthesis and metabolism of sterols and cholesterol. These findings suggest that MLPH, rich in amino acids and bioactive peptides, holds promise as a functional food ingredient for managing T2DM and developing antidiabetic foods.

BACKGROUND:Chronic obstructive pulmonary disease (COPD) is a common chronic respiratory disease and the third leading cause of death worldwide, with chronic inflammation as its primary pathogenesis. The dried leaves of Forsythia suspensa (Thunb.) have anti-inflammatory pharmacological activity and potential clinical promise for the treatment of COPD; however, its pharmacodynamic activity and mechanism of action remain to be clarified. RESULTS:In total, 82 compounds were identified from F. suspense leaf green tea extract (STW), mainly lignans, flavonoids, phenolic acids and phenylethanol glycosides, were identified in STW. STW alleviates enlargement of the alveolar sac and cavity, thickening of the alveolar wall and infiltration of inflammatory cells in COPD mice. In the model group, the contents of interleukin (IL)-6, tumor necrosis factor (TNF)-α, IL-8, IL-1β and malondialdehyde were decreased, and the levels of superoxide dismutase and catalase were increased in a dose-dependent manner (P < 0.05). Network pharmacological analysis identified 19 active STW components and 81 potential targets for the treatment of COPD. The key components include quercetin, ferulic acid, phillygenin, rutin and phillyri, whereas the core targets included TNF, protein kinase B alpha, epidermal growth factor receptor and metalloproteinase-9. Mainly through phosphatidylinositol 3-kinase/protein kinase B, calcium ions, nuclear factor-kappa B and other signaling pathways. CONCLUSION:STW affects multiple pathways and targets in the treatment of COPD, and mainly plays a role in alleviating pulmonary inflammation. © 2025 Society of Chemical Industry.

BACKGROUND:Type 2 diabetes mellitus (T2DM) poses a significant threat to global health, emphasizing the need for novel and effective treatments with minimal side effects. Anoectochilus roxburghii, commonly used in teas and food preparations, has shown promise as a functional food with antidiabetic activity. The present study aimed to investigate the bioactive composition and antidiabetic potential of A. roxburghii aqueous extract (AAE) on T2DM. RESULTS:The extraction method of AAE was optimized using response surface methodology, achieving a total flavonoid content of 3.24%. Forty-one components were identified in AAE using ultra performance liquid chromatography Q-Exactive Orbitrap (Thermo Fisher Scientific) mass spectrometry. Network pharmacology analysis predicted potential targets including NFKB1 and TLR4, suggesting the involvement of nuccear factor-kappa B and other related pathways. A T2DM model was induced by high-fat feeding and streptozotocin injection, showing increased glucose intolerance and insulin resistance, which AAE mitigated. AAE also modulated reactive oxygen species and cytokine levels, improved lipid profiles, glycogen and glucose transporter 2 levels. Hematoxylin and eosin and Oil Red O staining demonstrated that AAE alleviated pathological changes and lipid accumulation in the pancreas and liver of T2DM mice. Immunohistochemical observations further suggested that AAE may influence the expression of TLR4, NF-κB p65, MyD88 and IκBα. Among various physiological indicators, AAE improvement on 10 indices, including fasting blood glucose, fasting insulin and C-peptide, was comparable or superior to that of the positive drug metformin. CONCLUSION:These findings support the functional food potential of AAE in the dietary management of T2DM and provide preliminary evidence for its molecular targets and regulatory pathways. © 2025 Society of Chemical Industry.

Background Chinese dwarf cherry (Cerasus humilis) pomace is rich in proanthocyanidins, which exhibit antidiabetic activity.Objective This study investigates the protective effect of proanthocyanidins extracted from C. humilis pomace by regulating hepatic glucose-lipid metabolism in streptozotocin (STZ)-induced type 2 diabetic mellitus (T2DM) mice fed a high-fat diet (HFD).Methods The antidiabetic activities of Cerasus humilis pomace proanthocyanidins (CPPs) were evaluated through fasting blood glucose (FBG), oral glucose tolerance test (OGTT), and biochemical analysis, including AST, ALT, C-peptide, and hepatic glycogen in mouse serum, TC, TG, HDL-C, and LDL-C levels in the liver, as well as MDA and SOD levels. Thereafter, transcriptomic analysis, coupled with western blot and RT-qPCR, was employed to validate the molecular mechanism by which CPPs improved T2DM pathology in mice.Results CPPs significantly attenuated body weight gain, hyperglycemia, hepatic lipid accumulation, and liver injury in T2DM mice. Transcriptomics implicated the PI3K/Akt/FoxO1 and AMPK/ACC signaling pathways in mediating CPPs' anti-diabetic effects. Subsequently, western blot, immunohistochemistry, and RT-qPCR analyses confirmed that CPPs regulate these pathways and demonstrated significant suppression of key gluconeogenic enzymes, PEPCK and G6Pase.Conclusion CPPs can effectively ameliorate T2DM by suppressing hepatic glucolipid metabolism abnormalities, primarily through regulation of the PI3K/Akt/FoxO1 and AMPK/ACC signaling pathways.

Chinese dwarf cherry (Cerasus humilis) is a fruit unique to China, which is considered to have osteoprotective effects. However, no systematic experimental characterization was available. In this study, the osteoprotective activity and mechanism of Chinese dwarf cherry polyphenol extract (OPE) was studied. In vitro, OPE stimulated the alkaline phosphatase activity in the early differentiation stage, increased the osteocalcin level in the middle differentiation stage, and induced the formation of more bone-mineralized nodules in the late osteogenic stage. In vivo, OPE improved cancellous bone structure and maximum load of the femur in ovariectomized (OVX) rats. The balance between bone formation and resorption was regulated. Oxidative stress levels in the peripheral blood, liver and femur were reduced. OPE alleviated the disturbance in energy metabolism, muscle development, and muscle regulation-related signaling pathways caused by OVX and activated the calcium/adenosine monophosphate-activated protein kinase signaling pathway. Therefore, OPE is a potential dietary supplement for the prevention and treatment of osteoporosis.

This study investigated the combined therapeutic effects of Forsythoside A (FA) and Lonicera japonica polysaccharide (LP) on Alzheimer's disease (AD) through dual modulation of neuroinflammation and gut microbiota. HT22 cells were treated with A(325-35 to establish an AD model for examining the synergistic concentration ratio of FA and LP. An APP/PS1 double-transgenic AD mouse model was developed. The anti-AD activities of FA + LP were evaluated through behavioral testing, measuring amyloid-beta (A(3) and phosphorylated Tau (p-Tau) protein levels in the hippocampus and cortex, and assessing serum inflammatory factors. Transcriptomic analysis, coupled with quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and western blotting, was used to validate the molecular mechanism by which FA + LP improved AD pathology in mice. Finally, 16S rRNA sequencing technology was used to explore the anti-AD mechanisms of FA + LP from a gut microecology perspective. We found the optimal FA:LP synergistic ratio was 1:5. FA + LP treatment significantly improved cognitive function in behavioral tests while reducing A(3 and p-Tau accumulation in the hippocampus and cortex. Serum analysis showed decreased levels of pro-inflammatory cytokines (TNF-alpha, IL-1(3, IL-6). Mechanistic studies revealed FA + LP downregulated the IL-17-NF kappa B pathway, evidenced by reduced mRNA expression of Il-17a, Act1, I kappa ba, Nf kappa b p65, and Cox2, along with decreased IL-17A and NF kappa B p65 protein levels. Gut microbiota analysis demonstrated FA + LP increased beneficial bacteria (Lachnospiraceae and Muribaculum) while normalizing the Firmicutes/Bacteroidetes ratio. The combination therapy outperformed single treatments across all evaluated parameters. Our results demonstrate FA + LP (1:5) alleviates AD pathology through: suppressing neuroinflammation via IL-17-NF kappa B signaling inhibition, and restoring gut microbiota balance.

LA, one of important phenolic acids of health food Salvia miltiorrhiza Bge. and Crocus sativus L. (Saffron) has defined proangiogenic effect. However, the mechanisms of its proangiogenic effect are unknow. The present study aimed to further evaluate the proangiogenic effect of LA and to explore the potential mechanisms. The results showed that LA restored the length of PTK787-damaged intersegmental vessels (ISVs) and promoted the germination of subintestinal vessels (SIVs) in zebrafish. LA promoted the proliferation and migration of HUVECs under VRI-inhibited and normal conditions. The results of network pharmacology, transcriptomics, and molecular docking showed that ALB, AKT1, EGFR, SRC, and MMP9 were key targets for LA to promote angiogenesis. KEGG enrichment analysis and qPCR analysis showed that LA promoted angiogenesis by regulating the VEGF, PI3K-Akt and MAPK pathways. These results indicated that LA has proangiogenic effects both in vitro and in vivo by regulating the VEGF, PI3K-Akt and MAPK pathways.

Pruni Semen, the dried ripe seed of Prunus humilis, P. japonica, or P. pedunculata as recorded in the Chinese Pharmacopoeia, has been widely used in pharmaceutical and food industries. The adulteration of the marketed product with morphologically similar plants of the same genus has led to variable product quality and clinical effectiveness. This study systematically investigated the phylogenetic relationships, morphological traits, and chemical profiles of 37 Pruni Semen samples from planting bases, markets, and fields. DNA barcoding could successfully distinguish the genuine and counterfeit Pruni Semen, and the results indicated that there was almost no authentic Pruni Semen available in the market. The samples were divided into “big seed” (P. pedunculata and P. salicina seeds) and “small seed” (P. humilis, P. japonica, P. tomentosa, and P. avium seeds) categories based on morphology results. The notable discrepancy in the chemical characteristics of “big seed” and “small seed” was that “small seeds” were rich in flavonoids and low in amygdalin, whereas “big seeds” were the opposite. Furthermore, principal component analysis and clustered heatmap analysis verified the distinguishing features of “big seed” and “small seed” based on morphological and chemical characteristics. This study suggested that a combination of DNA barcoding and morphological and chemical characteristics can aid in the identification and quality evaluation of authentic and adulterated Pruni Semen. These findings may help standardize Pruni Semen available in the market and protect the rights and interests of customers.

In this study, the ultrasonic-assisted extraction process of proanthocyanidins from Cerasus humilis pomace was optimized using single-factor combined with orthogonal experiments. The in vitro antioxidant activity and hypoglycemic activity of the purified proanthocyanidins from Cerasus humilis pomace were determined using the DPPH radical scavenging test, ABTS+ radical scavenging test, FRAP method and α-glucosidase activity inhibition test. The extraction rate of proanthocyanidins was influenced in the following order:Sonication temperature>material-liquid ratio>sonication time>ethanol concentration. The best extraction process was that using 70% ethanol, sonication temperature of 60 ℃, sonication time of 40 min, and material-liquid ratio of 1:30 g/mL. Under these conditions, the yield of proanthocyanidins was 15.37 mg/g. The IC50 values of DPPH and ABTS+ radicals scavenged by purified proanthocyanidins were 0.10 mg/mL and 7 µg/mL, respectively, and the IC50 value for inhibiting α-glucosidase activity was 0.056 mg/mL. The extraction process was stable and feasible, the yield of proanthocyanidins was high, and the purified proanthocyanidins had strong in vitro antioxidant activity and hypoglycemic activity.

BACKGROUND:Multiflorin A (MA) is a potential active ingredient of traditional herbal laxative, Pruni semen, with unusual purgative activity and an unclear mechanism, and inhibiting intestinal glucose absorption is a promising mechanism of novel laxatives. However, this mechanism still lacks support and a description of basic research.PURPOSE:This study aimed to determine the main contribution of MA to the purgative activity of Pruni semen and elucidate the effect intensity, characteristics, site, and mechanism of MA in mice, and determine the novel mechanism of traditional herbal laxatives from the perspective of intestinal glucose absorption.METHODS:We induced diarrhoea in mice by administering Pruni semen and MA, and the defecation behaviour, glucose tolerance, and intestinal metabolism were analysed. The effects of MA and its metabolite on peristalsis of the intestinal smooth muscle were evaluated using an intestinal motility assay in vitro. Intestinal tight junction proteins, aquaporins, and glucose transporters expression were analysed using immunofluorescence; gut microbiota and faecal metabolites were analysed using 16S rRNA and liquid chromatography-mass spectrometry.RESULTS:MA administration (20 mg/kg) induced watery diarrhoea in over half of the experimental mice. The activity of MA in lowering peak postprandial glucose levels was synchronous with purgative action, with the acetyl group being the active moiety. MA was metabolised primarily in the small intestine, where it decreased sodium-glucose cotransporter-1, occludin, and claudin1 expression, then inhibited glucose absorption, resulting in a hyperosmotic environment. MA also increased the aquaporin3 expression to promote water secretion. Unabsorbed glucose reshapes the gut microbiota and their metabolism in the large intestine and the increasing gas and organic acid promoted defecation. After recovery, the intestinal permeability and glucose absorption function returned, and the abundance of probiotics such as Bifidobacterium increased.CONCLUSION:The purgative mechanism of MA involves inhibiting glucose absorption, altering permeability and water channels to promote water secretion in the small intestine, and regulating gut microbiota metabolism in the large intestine. This study is the first systematic experimental study on the purgative effect of MA. Our findings provide new insight into the study of novel purgative mechanisms.