Metformin-mediated modulation of gut microbiota-derived trimethylamine N-oxide (TMAO) in myocardial infarction: Insights from in vivo, metabolomics, and in silico studies.

Metformin drives the antibiotic resistome in activated sludge by reshaping microbial communities and promoting horizontal gene transfer.

A new sensitive, accurate and precise high-performance liquid chromatographic technique was developedbyemploying the Quality by Design (QbD) approach to quantify metformin (MET), dapagliflozin (DAPA), andsitagliptin (SITA). The mathematical models were developed using two independent factors, includingtheconcentration of the mobile phase and the flow rate. CCD was utilised to systematically explore the response surfacemethodology and to investigate how individual components influence the response variables. In the analytical process, a mobile phase consisting of acetonitrile and 0.1% formic acid in a 1:1 ratio was employed. It was providedat a flow rate of 1 mL/min while the column was kept at 30°C. With a correlation value (R2) of 0.999, the methodshowed excellent linearity within the concentration ranges of 40–60 µg/mL for MET, 9–12 µg/mL for SITA, and0.8–1.2 µg/mL for DAPA. The MET, SITA, and DAPA were found to have respective limits of detection (LOD) and quantification (LOQ) of 2.28 and 6.90 µg/mL, 0.55 and 0.045 µg/mL, and 0.015 and 1.66 µg/mL. The %drugrecovery was in the range of 98-100% and the %RSD of less than 2% for the HPLC method. The UVmethodwasalso linear with R² 0.999; LOD and LOQ were 12.12 and 36.74 µ/ml for MET, 0.8 and 2.5 µ/ml for SITA, 0.9and2.7 µ/ml for DAPA. "Using a Quality by Design (QbD) methodology, the results verified that the RP-HPLCmethodwas effectively optimised for the simultaneous quantification of MET, SITA, and DAPA. The UVand RP-HPLCtechniques both showed dependability in terms of accuracy, precision, linearity, and cost-effectiveness after beingvalidated in compliance with ICH Q2 (R1) guidelines.

ABSTRACTObjectiveDiabetic retinopathy (DR) is characterised by chronic neuroinflammation where the NLRP3 inflammasome plays a pivotal role. This study investigated the therapeutic potential and underlying mechanism of combining systemic metformin (MET) with intravitreal MCC950, a specific NLRP3 inhibitor, in a rodent model of DR.MethodsA type 2 diabetic rat model was induced by high‐fat diet and streptozotocin (STZ) injection. Diabetic rats were divided into DR, MET, MCC950 and MET+MCC950 treatment groups. Body weight and blood glucose were monitored. Retinal structural changes were assessed by HE and PAS staining. Apoptosis was detected by TUNEL assay, and oxidative stress was evaluated by ROS fluorescence. The expression and interaction of key proteins within the NEK7/NLRP3 pathway were analysed by Western blot and immunofluorescence.ResultsThe DR group exhibited significant retinal thinning, increased acellular capillaries, elevated apoptosis and oxidative stress. While monotherapies showed partial improvement, the MET+MCC950 combination yielded the most robust protective effects, nearly restoring retinal morphology and significantly reducing apoptosis and ROS levels. Mechanistically, combination therapy most effectively suppressed the activation of the NEK7/NLRP3 inflammasome pathway, as evidenced by decreased protein levels of NEK7, NLRP3, ASC, cleaved‐Caspase‐1 and IL‐1β. Immunofluorescence confirmed enhanced NEK7/NLRP3 interaction in DR, which was markedly inhibited by the combination treatment. A significant positive correlation was found between ROS levels and NEK7 expression.ConclusionThe study demonstrates that the combination of systemic metformin and intravitreal MCC950 confers superior protection against DR by synergistically inhibiting the NEK7/NLRP3 inflammasome pathway, resulting in reduced oxidative stress, apoptosis and inflammatory response. This novel combinational strategy presents a promising therapeutic approach for DR.

The roles of non-antibiotic pharmaceuticals in shaping the dissemination behaviours of antibiotic resistance genes (ARGs) in wastewater treatment systems remain poorly understood, and the influences of their transformation products have been overlooked. Here, we unveil more profound impacts of the metformin (MET) biotransformation product than the parent pollutant on the microbial community structure and ARG propagation of wastewater anaerobic sludge. The exposure to MET and its metabolic products guanylurea (GUA) at environmentally relevant concentrations both raised the methane production and resulted in up to 52.5% higher sludge ARGs abundance relative to the unexposed control. Especially, the GUA group showed up to 188-fold upregulation in several ARGs including bcrA, PmrF, acrB and mexF, enabled 3218-fold enrichment of plasmids from several bacteria. The underlying mechanisms were elucidated by integrated metagenomics, molecular dynamics simulations, and metabolic profiling analyses. MET and GUA were found to trigger coordinated cellular responses including disrupted glycerophospholipid metabolism, increased membrane permeability and broad metabolic reprogramming, which collectively boosted the ARGs dissemination. Overall, this work establishes a mechanistic link between micropollutant-induced microbial stress and ARGs propagation in anaerobic sludge, and advocates for re-evaluating the environmental risks of non-antibiotic pharmaceuticals and integrating resistance control into wastewater management framework.

Exploring molecular interactions of drugs in different biologically active solvents: A comprehensive review for safe and efficient drug delivery systems.

NIR/US-responsive injectable hyaluronic acid-based nickel sulfide hydrogel as a smart platform for synergistic antitumor therapy.

BackgroundThe leaves of Dimocarpus longan Lour. are used unilaterally as Chinese herbal medicines to treat diabetes in Chongzuo and Hezhou, Guangxi, but the mechanism of its treatment of diabetes is not yet clear, and further research is needed.MethodsThis study examined the effects of leaf ethanol extract of D.s longan Lour. on metabolic pathways and gut microbiota in rats with type 2 diabetes mellitus (T2DM). The rats were randomly divided into four groups: HG + HFD (T2DM model, fed with high‐sugar and high‐fat diet), control (regular diet), MET (positive metformin), and LYY (leaf ethanol extract of D. longan Lour). Metabolite profiles and gut microbiota composition were analyzed using liquid chromatography_mass spectrometry and 16S rDNA sequencing.ResultsMetabolomics analysis revealed 61 distinct metabolites in the LYY group, such as Leu‐Pro and taurolithocholic acid 3‐sulfate, which influence valine, leucine, and isoleucine metabolism, unsaturated fatty acid biosynthesis, fatty acid metabolism, bile secretion, and pyruvate and propanoate metabolism. Additionally, 16S rDNA sequencing showed that LYY significantly altered the abundance of gut microbiota such as Ligilactobacillus and Desulfobacterota (vs. HG + HFD group, p < 0.05).ConclusionLYY improved T2DM in rats may be associated with modulating metabolite levels and indirectly regulating glucose metabolism balance through changes in gut microbiota abundance. The efficacy of LYY in treating T2DM in rats may be related to the regulation of six metabolic pathways; it increased the abundance of Ligilactobacillus and Christensenellaceae _ R-7 _ group and decreased the abundance of Desulfobacterota, Colidextribacter, and Oscillibacter, thereby promoting impaired glucose tolerance and indirectly regulating the balance of glucose metabolism.

Introduction: Diabetes mellitus is a group of metabolic disorder in which there is high blood sugar over a prolonged period of time. This study aimed to compare gastrointestinal function and hepatic changes in diabetic rats treated with aqueous extract of Costus afer leaves, evaluate the effect of aqueous extract of C. afer leaves on liver enzymes in Wistar rats. Methods: Thirty-five male rats were grouped in 5 groups of 7 rats. To induce diabetes, 4mg/ml of streptozotocin, was injected at 50mg/kg body weight to animals in groups II, IV and V. Oven-dried C. afer leaves were crushed into powdered, and packaged separately and used to produce aqueous extract for the study. The extract was administered at a dosage of 800mg/kg body weight daily to rats in groups III and V. After 28 days, the animals were sacrificed and parameters were analyzed. A stock concentration of 20mg/ml of metformin was administered at a dose of 200mg/kg of body weight to the animal using oral gavage to group IV animals only. Extracted serum was used for Alkaline phosphatase (ALP), Alanine aminotransferase (ALT) and Aspartate aminotransferase (AST) analysis. Results: Treatment with C. afer alone (5.62±0.21), and in combination with metformin significantly reduced blood glucose compared to the untreated diabetic group (9.82±0.76). Final body weight was significantly reduced in diabetes mellitus group (DM) and DM+ Metformin (MTF) groups but improved with C. afer and DM+C. afer. AST, ALT, and ALP were significantly elevated in DM rats, indicating liver dysfunction. However, treatment with C. afer, especially in combination with metformin, significantly restored these enzymes toward normal levels. Conclusion: Findings support the antidiabetic, gastroprotective, and hepatoprotective efficacy of C. afer aqueous extract.

ABSTRACT A novel and reliable high‐performance liquid chromatography coupled with photodiode array (HPLC‐PDA) detection method was developed and validated for the simultaneous determination of metformin hydrochloride, its related impurity cyanoguanidine, and five co‐formulated antidiabetic agents—linagliptin, vildagliptin, sitagliptin, empagliflozin, and dapagliflozin—in pharmaceutical dosage forms. Chromatographic separation was achieved on a PerkinElmer Analytical Cyano column (150 × 4.6 mm, 5 µm) using a gradient elution of 15 mM orthophosphoric acid (containing 5 mM sodium heptane sulfonate) and acetonitrile at a flow rate of 1.0 mL/min. The detection wavelengths were set individually for each analyte (205–228 nm) using a PDA detector. The method was validated according to the International Council for Harmonization Q2(R2) guideline. Linearity was demonstrated within the ranges of 100–600 µg/mL for metformin, 0.2–1.0 µg/mL for cyanoguanidine, 0.25–1.25 µg/mL for linagliptin, 5–30 µg/mL for vildagliptin, 5–30 µg/mL for sitagliptin, and 0.5–3.0 µg/mL for both empagliflozin and dapagliflozin, with correlation coefficients (R 2 ) greater than 0.99. Recovery values were between 98% and 102%, and the percentage relative standard deviation values were below 2%, confirming the accuracy, precision, and overall analytical performance. No interference was observed from excipients or blank matrices, and robustness tests confirmed the method's stability under slight variations in analytical conditions. The proposed method was successfully applied to the analysis of commercial formulations, demonstrating its suitability for routine quality control and stability testing of metformin‐containing products and their fixed‐dose combinations with dipeptidyl peptidase‐4 and sodium‐glucose cotransporter‐2 inhibitors.

Abstract Castration-resistant prostate cancer (CRPC) is an aggressive disease with a high propensity for metastasis and limited therapeutic options. This study investigates the potential of a combined regimen using Ganoderma lucidum polysaccharide (GLP), a bioactive food component derived from the edible Linzhi mushroom, and metformin (MET) in the treatment of CRPC, with a particular focus on metastasis and intestinal barrier function. We found that GLP and MET synergistically inhibited the migration of CRPC cells (PC3, DU145) in vitro. The combination treatment profoundly suppressed epithelial–mesenchymal transition (EMT), as evidenced by increased E-cadherin and decreased N-cadherin, Snail, MMP2, and MMP9 levels. In a mouse model of metastasis using tail vein injection of PC3-luc cells, the GLP/MET combination significantly reduced the incidence and burden of multiorgan and bone metastases, as quantified by bioluminescence imaging, and extended tumor-free survival. Interestingly, from a food safety and health perspective, while MET monotherapy failed to protect the gut, GLP alone and co-administration effectively preserved intestinal barrier integrity. This was demonstrated by the restoration of goblet cells, increased MUC2 expression, and upregulation of tight junction proteins (ZO-1 and occludin), alongside a reduction in macrophage-driven inflammation in colon tissues. Our findings indicate that GLP not only enhances the antimetastatic efficacy of MET but also ameliorates concomitant intestinal damage. This study highlights the promise of GLP as a safe dietary adjuvant to conventional therapeutics, offering a strategy to improve CRPC management while enhancing gastrointestinal tolerance.

Diabetic nephropathy is a serious and common complication among patients with both type 1 and type 2 diabetes, and it significantly reduces the patient’s quality of life. This study aimed to assess the reno preventive effects of valproate sodium (VPS) and metformin (MET) on alloxan-induced diabetic nephropathy and to elucidate their mechanisms of action, ‘type 1 diabetic mice’ (25–30 g) were established using a single dose of alloxan (‘120 mgkg-1’). and the diabetic mice were treated with three doses of VPS (10, 20, and 40 mg/kg) and MET (200 mg/kg) for a period of 28 days. Specific tests were performed to evaluate inflammatory gene expression (TNF-α, IL-6, and NF-κB) and histopathological changes and apoptotic factors (Bax/Bcl2, Caspase3). Our results have shown, VPS and MET led to significant decreases in blood glucose levels, thereby reflecting the improvement of impaired kidney function and decreasing elevated renal mRNA levels of inflammatory genes (TNF-α, IL-6, and NF-κB) in diabetic mice. A significant increase in the expression of be ‘Sirt1 and Bcl-2’ and decrease in (TNF-α, IL-6, and NF-κB) was observed in the kidneys of diabetic mice receiving MET/VPS Moreover, MET/VPS successfully prevented diabetes induced ‘histopathological deleterious changes’ in the kidneys of mice so it can concluded that MET and VPS alone or in combination can prevent alloxan-induced diabetic nephropathy through attenuating inflammatory markers and probably with suppression of apoptosis.

ABSTRACT: The objective of this study was to formulate, optimize, and evaluate bilayered floating tablets containing dapagliflozin and metformin hydrochloride for sustained drug release and improved glycemic control compared with a marketed innovator product. Combination therapy of dapagliflozin, a sodium-glucose co-transporter-2 inhibitor, and metformin, a biguanide, provides effective management of type 2 diabetes mellitus but often requires multiple daily doses. To enhance therapeutic efficiency and patient compliance, a bilayered dosage form combining immediate and sustained release was developed. The formulation consisted of an immediate-release (IR) layer of dapagliflozin for rapid onset of action and a sustained-release (SR) layer of metformin HCl to maintain prolonged plasma concentration. Various batches (B7–B21) were prepared using different gas-generating agents and release-retarding polymers such as Hypromellose (HPMC K100M) and Carbopol 934P. Optimization was carried out to assess the effects of excipient concentration on key parameters including hardness, friability, floating lag time, total floating duration, and in vitro drug release. Among all formulations, Batch B17 showed superior performance with hardness of 5.4 kg/cm², friability of 0.19%, floating lag time of 60 seconds, and total floating duration of 12 hours. The in vitro release profile demonstrated 99.83% dapagliflozin release within 30 minutes and sustained metformin HCl release up to 12 hours (99.31%), closely comparable to the marketed product (XIDUGO XR). The combination of HPMC K100M and Carbopol 934P provided excellent matrix stability and prolonged gastric retention. In conclusion, the optimized bilayered floating tablet (B17) effectively combines immediate and sustained release profiles, reducing dosing frequency and improving patient adherence in diabetes management. Further in vivo, preclinical, and clinical studies are required to establish bioequivalence and confirm therapeutic performance.

The Aim of this present study was to develop an oral sustained release matrix tablet of metformin HCl and evaluate the influence of Xanthum gum on the drug release profile. The use of FTIR for drug – excipient compatibility is appropriate. The wet granulation method is suitable for matrix tablet preparation, incorporating hydrophilic HPMC K100M, PVP K30, and Xanthan gum as controlling agents for the release rate. Lactose served as the filler. The physicochemical tests (weight variation, hardness, friability, content uniformity, and in vitro dissolution) were standard and appropriately listed. Both pre- and post-compression parameters are good that both stages were evaluated in this study. The data on drug release were analyzed using multiple models to determine the kinetics and mechanism of drug release. A clear comparison between HPMC alone and HPMC – Xanthum gum combination was made. Quantify the difference in drug release, HPMC alone released 90% of drug within 6 hours, while the combination extended release to 12 hours. Release kinetics following Zero order kinetics with Higuchi model diffusion-controlled release mechanism. Formulation F8 was identified as the optimal formulation. The dissolution profile of formulation F8 can be characterized by Release kinetics following Zero order kinetics with Higuchi model diffusion-controlled release mechanism. Keywords: Matrix tablets, Metformin hydrochloride, Xantham gum, HPMCK100, PVPK30

Abstract Abstract: Introduction Metformin is used in adults with diabetes and by pregnant women. However, metformin crosses the placenta and may restrict fetal growth. In primary fetal hepatocytes, we demonstrated that metformin decreases O2 consumption and glucose production, consistent with its effects in adults, yet induces fetal-specific metabolic stress. Objective Determine whether metformin, in combination with Western diet (WD), impairs oxidative metabolism, injuring the fetal liver. Methods Pregnant Rhesus macaques received vehicle (VEH) or metformin (MET) and a healthy control diet (CD) or WD (n = 10/group) from 0.2 to 0.85 gestation. After cesarean delivery, cord blood samples, fetal weights, and isolated fetal hepatocytes were obtained. Results Hepatocytes from MET vs. VEH fetuses had higher O2 consumption rates with greater maximal (P = 0.015) and spare respiratory capacity (P = 0.05). There were tendencies for increased MFN2 protein (P = 0.09) and PGC1A gene expression (P = 0.06) in liver tissue, indicating increased mitochondrial fusion and biogenesis. MET-exposure did not affect WD-induced liver triglyceride accumulation nor cord blood glucose or O2 levels. However, histology showed increased collagen with MET and WD exposure. Furthermore, WD+MET fetuses had increased blood lactate ­concentrations. Fetal weight was similar across groups; however, CD+MET versus CD+VEH female fetuses weighed less (interaction, P = 0.02). Conclusions Metformin is associated with fetal metabolic stress, resulting in mitochondrial remodeling and increased oxygen consumption capacity. Further, increased lactate may underlie ­collagen deposition and hepatocellular injury. Ongoing studies are underway to understand if liver metabolic effects are linked with sex-dependent growth effects. (Supported by NIH-R01-DK128187)

Based on improved thin-film dispersion method with an optimized preparation process, elevated encapsulation efficiency and excellent stability of the single-chamber liposome, lip@MET/CUR, were achieved for co-delivery of metformin (MET) and curcumin (CUR). To increase the volume of the hydration chamber and the specific surface area during lipid membrane formation, Tween-80 and glass microspheres were introduced in the preparation process. On this basis, the optimal process parameters for high encapsulation efficiency were screened and determined by combining the analytic hierarchy process (AHP), entropy weight method (EWM), and Box-Behnken response surface optimization. Eventually, the optimal encapsulation efficiencies for MET and CUR were determined to be 46.4% ± 1.3% and 94.1% ± 1.5%, respectively. The lip@MET/CUR exhibited an average particle size of 150 ± 2.5 nm with uniform particle size and good storage stability. In vitro drug release experiments revealed a significant sustained-release characteristic of lip@MET/CUR. Specifically, the cumulative release rate of MET decreased from 96.8% to 57.4% within the initial 2 h. Results from MTT assays and experiments conducted in tumor-bearing mice further demonstrated that lip@MET/CUR was more effective in inhibiting the growth of HepG2 cells and tumors compared to free CUR or lip@CUR. In summary, our findings suggest that the optimized lip@MET/CUR formulation holds great potential as a candidate for investigating the synergistic effects of CUR and MET in tumor treatment.

BackgroundPolycystic ovary syndrome (PCOS) affects both obese and normal-weight women, with limited treatment options available for the nonobese population. Metformin (MET), an insulin sensitizer, is used to ameliorate insulin resistance and associated reproductive endocrine metabolic dysfunctions in PCOS patients. The efficacy of chiglitazar, a peroxisome proliferator-activated receptor (PPAR) pan-agonist used for type 2 diabetes treatment, is undefined in PCOS patients. In this randomized controlled trial, the effects of metformin versus chiglitazar on insulin resistance and reproductive endocrine metabolism are assessed in normal-weight women with PCOS.MethodsFifty-five normal-weight women with PCOS aged 18 to 45 years were included. Patients were randomly assigned to receive either chiglitazar (32 mg once daily) or MET (500 mg twice daily). Anthropometric measurements, menstrual cycle changes, sex hormone characteristics, and an oral glucose-insulin release test (OGIRT) were performed after three months of continuous use.ResultsFollowing 12 weeks of treatment with chiglitazar, there were notable improvements in insulin and blood glucose levels at the 120-minute mark of the OGIRT, and the peak insulin levels were significantly earlier than those at baseline, indicating a more pronounced effect than that in the MET group. Moreover, both the chiglitazar and MET treatments led to significant improvements in menstrual cyclicity, and luteinizing hormone (LH) and testosterone (Testo) levels, with no significant differences detected between the two groups. Prolactin (PRL) levels were significantly elevated in the Chiglitazar group compared with the MET group. There was also no significant difference in the efficacy of the two treatments for PCOS in subgroups with different baseline IR0 values.ConclusionIn normal-weight PCOS patients, chiglitazar is similar to MET with regard to improving menstrual frequency and total testosterone (TT) and LH levels. Compared with MET, chiglitazar significantly improves fasting insulin levels, insulin and blood glucose levels at 120 min of the OGIRT and advances the insulin peak.Trial registrationThis single-center, open-label, 1:1 randomized controlled trial is registered with ClinicalTrials (NCT06125587, ClinicalTrials.gov) on 2023-11-05.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12902-025-02082-0.

To study the mechanism of arsenic trioxide (ATO) combined with metformin (MET) in promoting apoptosis of leukemia cells. CCK-8 method was used to detect the viability of leukemia cell line KG1a, K562, and THP1 cells treated by ATO monotherapy, MET monotherapy, and ATO combined with MET. Flow cytometry was used to detect cell cycle and apoptosis. RT-qPCR was used to detect the mRNA expression of PI3K/Akt and LKB1/AMPK pathway-related genes. Western blot was used to detect the expression of PI3K/Akt and LKB1/AMPK pathway-related proteins and autophagy-related protein LC3B and P62. Compared with the ATO monotherapy group, ATO combined with MET significantly inhibited the growth of KG1a, K562 and THP1 cells, and the difference in KG1a cells was more statistically significant. The combination of the two drugs induced KG1a cell cycle arrest, promoted apoptosis, increased the expression of autophagy-related protein LC3B and P62, up-regulated the mRNA expression levels of PI3K/Akt pathway and LKB1/AMPK pathway-related genes, as well as the expression of LKB1/AMPK pathway-related proteins, and down-regulated the expression of PI3K/Akt pathway-related proteins. ATO combined with MET promotes apoptosis by up-regulating LKB1/AMPK and down-regulating PI3K/Akt signaling pathway to regulate the autophagy of leukemia cells.

BackgroundAlzheimer's Disease (AD) and type 2 diabetes mellitus (T2D) share an exacerbated neuroinflammatory response coordinated by microglial cells and a decreased autophagic flux. Treatment with the normoglycemic metformin (MET) has been associated with reduced microglial activation and restored autophagy in T2D models. Our objective, within the framework of drug repositioning, is to evaluate the therapeutic potential of MET on experimental AD using mouse models and in vitro experiments.MethodWe treated 9‐month‐old male PDAPP‐J20 mice carrying AD mutations with MET (350 mg/kg i.p. three times a week for three weeks) or vehicle and we assessed cognitive profile using spatial memory tests such as the Barnes maze. From brain sections containing hippocampus, we analyzed the impact of MET on amyloid pathology, neurogenesis status by evaluation of the new‐born neuron marker DCX and microglial status. In vitro, the murine BV2 microglial cell line was exposed to 0.05 µM of Aβ 1‐42 for 24 hours, followed by 1 hour of treatment with 0.2 mM MET, and autophagic flux was evaluated under these conditions.ResultIn MET‐treated mice we found an improvement in spatial memory and hippocampal neurogenesis and a decrease in amyloid pathology compared to vehicle‐treated group. AD mice exhibited a decrease in the homeostatic microglial marker TMEM119, which was reversed by MET treatment. Additionally, a restoration of microglial autophagic flux was observed, evidenced by a reduction in the autophagosome marker p62 in transgenic mice treated with MET. In vitro, exposure to amyloid peptides resulted in a blockade of autophagic flux, assessed by p62 accumulation using Bafilomycin, which was reversed by MET.ConclusionOur results indicate that metformin may have a therapeutic effect in AD models restoring cognitive performance. This effect appears to be due to a microglial gain of function by rehabilitating the autophagic flux blocked by amyloid peptides. Metformin may also contribute to the reduction of amyloid pathology and improvement of hippocampal neurogenesis.

Metformin attenuates placental oxidative stress through Nrf2/Keap1 signaling in preeclampsia rats.