OBJECTIVE:To assess the association between Colquhounia Root Tablet (CRT), a Tripterygium hypoglaucum-derived immune-metabolic modulator, and kidney outcomes in type 2 diabetic kidney disease (DKD) patients with heavy proteinuria despite guideline-directed therapy (renin-angiotensin system blockade, SGLT2 inhibitors, finerenone). METHODS:A retrospective propensity score-matched cohort study was conducted using hospital-based electronic health records. Patients with heavy-proteinuric type 2 DKD (UACR > 300 mg/g) were matched 1:1 (159 pairs). The primary endpoint was major adverse kidney events (MAKE: ≥57% sustained eGFR decline, end-stage renal disease, or kidney-related death), analyzed using multivariable-adjusted flexible parametric survival models. RESULTS:The matched cohort had a median UACR of 1818 mg/g; 40.2% were classified as KDIGO very high risk. Over 5 years, CRT use was associated with lower MAKE incidence (52.6% vs 70.9%; adjusted HR 0.60; 95% CI, 0.43-0.85; p = 0.004), slower annual eGFR decline (-5.08 vs - 7.05 mL/min/1.73 m2/year; p < 0.001), and greater UACR reduction at 12 months (-52.8% vs 1.8%; p < 0.001), with comparable safety. CONCLUSION:Adjunctive CRT was associated with improved kidney outcomes in high-risk heavy-proteinuric DKD, supporting future randomized trials targeting immune-metabolic phenotypes with persistent proteinuria and residual risk.

The pathogenesis of diabetic kidney disease (DKD) is complex and closely related to ferroptosis and immune dysregulation, but the relevance is unclear. The present study investigates the potential mechanisms of ferroptosis-related genes (FRGs) in DKD and their relationship with the immune-inflammatory response. It searches for new diagnostic biomarkers to help diagnose and treat DKD. Four Gene Expression Omnibus (GEO) datasets, GSE30528, GSE30529 and GSE30122 as the test set, and GSE96804 for validation, were analyzed. FRGs were obtained from GeneCards, and 47 ferroptosis-related differentially expressed genes (FRDEGs) were identified by intersecting with DKD-related differentially expressed genes. Functional enrichment analyses, including Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, Gene Set Enrichment Analysis and Gene Set Variation Analysis, revealed that these FRDEGs are primarily associated with ferroptosis, hypoxia response and immune inflammation. Subsequently, the weighted gene co-expression network analysis (WGCNA) was employed to expand the ferroptosis-related gene network, and intersection of the 47 FRDEGs with key WGCNA module genes yielded 10 key genes. Based on the 10 key genes, the least absolute shrinkage and selection operator and support vector machine algorithms identified three hub genes [chemokine ligand 5 (CCL5), forkhead box C1 (FOXC1) and lactotransferrin (LTF)] for DKD diagnosis. Receiver operating characteristic curves confirmed their diagnostic value, with FOXC1 and LTF validated in the independent dataset. Immune infiltration analysis via CIBERSORT revealed eight immune cell types with significantly different infiltration levels between the DKD and control group in the integrated GEO datasets. Notably, both LTF and CCL5 showed a significant positive correlation with gamma delta T cells (γδT). Quantitative PCR results confirmed differential expression of the three hub genes in the DKD group, with elevated expression observed in DKD mice following intervention with rosiglitazone and hyperoside.

The pathogenesis of diabetic kidney disease (DKD) is diverse and complex, and there is no particularly effective drug therapy. Qikui granules (QKG) is a traditional Chinese medicine compound preparation. Previous clinical studies have demonstrated that QKG has good anti-DKD effects. The present study aimed to further investigate the efficacy and mechanism of QKG in the treatment of DKD using the multi-omics strategy of metabolomics, lipidomics and 16S rRNA sequencing. Firstly, the pharmacological mechanisms of QKG intervening DKD were explored by metabolomics and lipidomics. Subsequently, antibiotic treatment was used to construct a pseudo-sterile model to validate the potential mechanism of QKG by modulating microbiota. Metabolomics and lipidomics analysis showed that QKG significantly reversed the upregulation of 26 special potential biomarkers in feces, including phosphatidylcholines (PC), phosphatidylethanolamines (PE) and unsaturated fatty acids, with glycerophospholipids and arachidonic acid being the most relevant metabolic pathways. The pseudo-sterile mice model validated the microbiota-dependent therapeutic effect of QKG on DKD from the perspective of both pharmacodynamic and gut microbiota. Besides, the correlation analysis showed that 26 special potential biomarkers characterizing the efficacy of QKG against DKD were negatively correlated with Candidatus_Arthromitus. Collectively, our findings demonstrated that QKG may exert microbiota-dependent efficacy in DKD-associated metabolism disorders and inflammation, mainly through upregulating Candidatus_Arthromitus, and then reducing PC, PE, and unsaturated fatty acids.

BACKGROUND Solute carrier family 30 member 8 (SLC30A8) is a protein encoded for a zinc efflux transporter in insulin secretory granules of pancreatic islets. Genome wide association study has demonstrated that SLC30A8 is a susceptibility gene for type 2 diabetes (T2D). AIM To evaluate whether DNA methylation changes in SLC30A8 is associated with T2D and diabetic kidney disease (DKD). METHODS A total of 820 Chinese subjects, including non-diabetic control subjects, newly diagnosed T2D, T2D with and without DKD and additional 230 healthy subjects before and after physical exercise were enrolled in this study. DNA methylation levels of six CpG sites in the SLC30A8 gene promoter were analyzed with bisulfite pyrosequencing technique. RESULTS DNA methylation levels of SLC30A8 were found to be remarkably high (81.01% +/- 3.9%) in this Chinese cohort. Compared to non-diabetic controls, SLC30A8 promoter methylation levels in T2D and DKD patients were significantly higher (81.7% +/- 3.9% and 81.6% +/- 3.8% vs 79.4% +/- 2.5%, P < 0.001 and P < 0.001). Mendelian randomization analysis implicated a causal relationship between decreased SLC30A8 expression and higher risk of T2D. We further investigated SLC30A8 promoter methylation changes in 230 healthy subjects before and after physical exercise and found that the specificity of SLC30A8 promoter hypermethylation was not influenced by environmental factors such as physical exercise. CONCLUSION The current study provides evidence that SLC30A8 gene promoter region exhibits hypermethylation. The SLC30A8 promoter hypermethylation is associated with both T2D and DKD in a Chinese population but not interfered by physical intervention. The findings suggest that SLC30A8 may have its potential value as a novel target of epigenetic pharmacology.

Diabetic kidney disease (DKD) has a complex and multifactorial pathogenesis, and highly effective pharmacotherapies for this condition remain limited. Qikui granules (QKG), a compound traditional Chinese medicine preparation, has shown favorable therapeutic efficacy against DKD in clinical studies. Our prior work, based on integrated network pharmacology, 16S rRNA gene sequencing, and lipidomic profiling of serum and kidney tissues, demonstrated that QKG ameliorates DKD in db/db mice by enriching Candidatus Arthromitus and modulating lipid homeostasis. As a follow-up validation of our earlier findings, the present study adopts an integrated multi-omics strategy combining fecal metabolomics, lipidomics, and 16S rRNA sequencing, coupled with a pseudo-germ-free mouse model, to further elucidate the therapeutic efficacy of QKG against DKD and the pivotal role of gut microbiota in its renoprotective effects. Firstly, fecal metabolomic and lipidomic profiling was performed to explore the pharmacological mechanisms of QKG against DKD. Subsequently, a pseudo-germ-free mouse model established via antibiotic administration was used to verify the microbiota-mediated mechanism of QKG. Metabolomic and lipidomic analysis revealed that QKG significantly reversed the upregulation of 24 special potential biomarkers (SPBs) in feces, including phosphatidylcholines (PC), phosphatidylethanolamines (PE) and unsaturated fatty acids (UFA); glycerophospholipid metabolism and arachidonic acid metabolism were identified as the most relevant pathways. The pseudo-germ-free mouse model validated the microbiota-dependent therapeutic effect of QKG on DKD from both pharmacodynamic and gut microbiota composition perspectives. Furthermore, correlation analysis showed that the 24 SPBs associated with the anti-DKD efficacy of QKG were negatively correlated with the relative abundance of Candidatus Arthromitus. Collectively, our findings suggest that QKG exerts microbiota-dependent efficacy against DKD-related metabolic disorders and inflammation, mainly by increasing the abundance of Candidatus Arthromitus and thereby reducing the levels of PC, PE, and UFA.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a global liver disorder with a rising incidence. Early-to-middle-stage MASLD remains amenable to clinical intervention; without timely management, it may progress to fibrosis and cirrhosis. Therefore, identifying novel targets through integrated multi-technical approaches is critical for preventing and treating MASLD. We developed an integrative multi-omics and spatial proteomic strategy combined with in vitro validation. CyTOF was applied to high-dimensional immunophenotyping of the MASLD liver microenvironment. Key cell populations were then sorted accordingly for mass spectrometry to uncover their core signaling. Focusing on identified key amino acid metabolic enzymes, we performed IMC for spatial single-cell proteomic profiling to localize metabolic aberrant subsets. Finally, functional phenotypes and mechanisms were validated in cell models. CyTOF revealed significant expansion of myeloid-derived cells. Thus, by targeting myeloid-derived cells, proteomic and phosphoproteomic analyses identified two severely impaired amino acid metabolic pathways: the glycine metabolic pathway regulated by AGXT2 and the proline metabolic pathway regulated by PYCR3. We then focused on AGXT2 and PYCR3 for spatial exploration via IMC and ultimately identified a macrophage subset negative for both AGXT2 and PYCR3. These macrophages were significantly elevated within MASLD and exhibited high spatial colocalization with inflammatory cells and fibrotic cells. Compared with dysregulated M1 subsets (M1-C3), dysregulated M2 subsets (M2-C1) indicated stronger pro-inflammatory and pro-fibrotic potential. In vitro models using human and murine cells confirmed that AGXT2⁻PYCR3⁻ macrophages exhibited enhanced proliferation and migration, secreted higher levels of inflammatory cytokines, chemokines, and classic fibrotic proteins, and exerted strong inductive effects on hepatic fibrotic cells, with downregulation of intercellular GSH. Mechanistically, the NF-κB and MAPK/AP-1 pathways were involved in inflammatory responses, while the p-SMAD3(T8)/TGFβ pathway contributed to the fibrotic phenotype. However, the abnormal biological functions could be effectively rescued by supplementation with corresponding amino acids, accompanied by the reversal of aberrant signaling pathway activation. In summary, our study explores an initial link between amino acid metabolism and early-to-middle-stage MASLD. AGXT2⁻PYCR3⁻ macrophages are significantly enriched in MASLD with pro-inflammatory and pro-fibrotic potential, and amino acid supplementation can rescue their aberrant phenotypes in vitro, warranting further validation of their treatment potential.

Background Myricetin, a bioactive flavonoid from Abelmoschus manihot, has demonstrated therapeutic potential for metabolic dysfunction-associated steatotic liver disease (MASLD) and its progressive form, metabolic dysfunction-associated steatohepatitis (MASH). However, its precise mechanisms, particularly concerning mitochondrial homeostasis, remain inadequately elucidated. Objectives The present study evaluated the therapeutic efficacy of myricetin in alleviating hepatic steatosis, inflammation, fibrosis, and insulin resistance associated with MASLD/MASH, with a specific focus on unraveling the role of mitophagy regulation. Methods MASLD and MASH models were established in mice using a high-fat diet (HFD) or a Gubra-Amylin NASH (GAN) diet, followed by myricetin treatment. Systemic metabolism, liver injury, histology, and insulin sensitivity were assessed. Transcriptomic profiling was performed to analyze metabolic pathways. Molecular docking, surface plasmon resonance (SPR), co-immunoprecipitation, and immunofluorescence were used to study the interaction between myricetin and PINK1 and its impact on PINK1/Parkin-mediated mitophagy. Finally, in vitro loss-of-function experiments using shPINK1 were conducted to validate the mechanism. Results Myricetin significantly ameliorated hepatic steatosis, inflammation, fibrosis, and systemic insulin resistance in MASLD/MASH mice. Transcriptomics revealed enhanced fatty acid β-oxidation and mitochondrial function. Mechanistically, myricetin directly bound to PINK1, inhibiting its mitochondrial import through the TOM complex (TOM40) and subsequent cleavage by the PARL protease, thereby stabilizing PINK1 on the outer mitochondrial membrane. This stabilization activated PINK1/Parkin-dependent mitophagy, restoring mitochondrial integrity. Notably, the myricetin-mediated improvements in mitophagy and mitochondrial function were negated by PINK1 silencing. Conclusion Myricetin mitigates MASLD/MASH progression by acting as a novel PINK1 stabilizer, augmenting PINK1/Parkin-dependent mitophagy to enhance mitochondrial quality. This study highlights myricetin as a potent intervention targeting mitochondria to combat metabolic liver diseases.

BACKGROUND Lianhe Xiaozhi ointment (LXO), an innovative formulation derived from the classic Huanglian Wendan decoction, has been granted a national invention patent. With increasing years of clinical expertise within the Jiangsu Province Hospital of Chinese Medicine in China, LXO has become increasingly recognized as a potent remedy for metabolic disorders, particularly metabolic dysfunction-associated steatotic liver disease (MASLD). To date, specific bioactive components and underlying mechanisms remain unclear. AIM To determine the bioactive components of LXO and clarify its mechanisms of action relevant to management of MASLD. METHODS We used ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry and network pharmacology approaches to systematically determine the key bioactive components of LXO and elucidate biological pathways modulated in the treatment of MASLD. Critical signaling pathways were also elucidated via hepatic transcriptomic analysis, and an exhaustive and rigorous exploration of the therapeutic efficacy and underlying mechanisms of LXO in MASLD was conducted via a combination of in vivo and in vitro high-fat model experiments. RESULTS Network pharmacology analysis revealed six pivotal bioactive components within LXO that collectively serve as the cornerstone for their efficacy against MASLD. LXO exerts multiple therapeutic effects, including weight gain retardation, amelioration of glucose and lipid metabolism disturbances, liver injury mitigation, hepatic inflammation alleviation, and correction of gut microbiota disorders. Multiple platform analyses, including hepatic transcriptomics, quantitative real-time polymerase chain reaction, and western blotting, confirmed that LXO induces peroxisome proliferator-activated receptor alpha (PPARα) transcriptional activation in the liver. In vitro studies confirmed that a PPARα antagonist markedly diminishes the lipid-lowering capacity of LXO. CONCLUSION Collectively, our findings suggest that LXO exerts therapeutic effects on high-fat diet-induced MASLD at least in part via PPARα pathway activation.

OBJECTIVE:In this study, the causation between serum metabolites and the risk of Diabetic Nephropathy (DN) was investigated by means of a Mendelian Randomization (MR) analysis. METHODS:Our data on diabetic nephropathy were obtained from the IEU OpenGWAS Project database, while serum metabolite data originated came from the GWAS summary statistics by Chen et al. The Inverse Variance Weighted (IVW) method was the main analysis approach, with Weighted Median (WME) and MREgger regression serving as supplementary approaches to construing the causalities between serum metabolites and the DN risk. In addition to the MR-Egger regression intercept, Cochran's Q test was utilized for sensitivity analysis, with P values used as the metric to assess the results. RESULTS:In total, 14 SNPs regarding serum metabolites were chosen as Instrumental Variables (IVs). The IVW results indicated that levels of Behenoylcarnitine (C22), Arachidoylcarnitine (C20), and the ratio of 5-methylthioadenosine (MTA) to phosphate exerted a positive causal effect on the DN risk. Conversely, levels of 5-hydroxylysine, Butyrylglycine, 1-stearoyl-glycerophosphocholine (18:0), Isobutyrylglycine, 1-stearoyl-2- oleoyl-GPE (18:0/18:1), N2,N5-diacetylornithine, 2-butenoylglycine, 3-hydroxybutyroylglycine, N-acetylisoputreanine, the ratio of Arginine to Ornithine, and the ratio of Aspartate to Mannose exerted a negative impact of causality on the DN risk. By identifying these serum metabolites, high-risk patients can be recognized in the early stages of diabetic nephropathy, enabling preventive measures or delaying its progression. These findings also provide a solid foundation for further research into the underlying etiology of diabetic nephropathy. CONCLUSION:The translation of serum metabolites into clinical applications for DN aims to utilize changes in serum metabolites as biomarkers for early diagnosis, thereby monitoring the progression of DN and providing a foundation for personalized treatment. For instance, the development of serum metabolite diagnostic kits could be used for early detection and prevention of DN. Changes in metabolites can help identify different stages of DN.

While the functional adaptation of β-cells during type 2 diabetes progression is well-established, the role of non-β islet cells remains largely unexplored. Utilizing single-cell RNA sequencing, we identified a substantial expansion of the macrophage population and a concomitant reduction in the proportion of mesenchymal stem cells (MSCs) within the islets of diabetic mice transitioning from metabolic compensation to decompensation. Under conditions of metabolic stress, macrophages extensively infiltrated the islets and adopted a pronounced pro-inflammatory phenotype. This phenotypic shift impaired β-cell glucose-stimulated insulin secretion and induced β-cell apoptosis. Simultaneously, macrophage-derived inflammatory factors, notably TNF-α, suppressed MSC proliferation and downregulated Wntless (Wls), thereby reducing extracellular Wnt transport. The resultant loss of Wls diminished MSCs' capacity to provide trophic support to β-cells and hindered the transition of macrophages to an anti-inflammatory phenotype. This self-perpetuating cycle establishes a chronic pro-inflammatory environment within the islets, culminating in β-cell functional deterioration and the onset of diabetes. Experimental intervention involving macrophage elimination and MSC administration was shown to disrupt this detrimental cycle, restoring β-cell function and glycemic control. Collectively, our findings reveal that macrophages and MSCs jointly govern β-cell adaptation through intricate paracrine crosstalk. Modulating these macrophage-MSC interactions holds significant therapeutic implications for maintaining β-cell integrity and underscores the considerable potential of MSC-based therapies for type 2 diabetes treatment.

Diabetic kidney disease(DKD),one of the most severe microvascular complications of diabetes,has become the leading cause of chronic kidney disease and end-stage kidney disease in China. Branched-chain amino acid(BCAA),essential amino acid in humans,constitutes an important part of amino acid metabolism. Accumulating evidence indicates that abnormal BCAA metabolism plays a critical role in the initiation and progression of DKD and is closely related to the injury of glomerular podocytes and renal tubular epithelial cells,whereas the relevant pathogenic mechanisms have not been systematically summarized. This review elaborates on BCAA metabolic pathways and the mechanisms by which disturbed BCAA metabolism induces renal damage in DKD,and discusses the therapeutic potential of BCAA and their key metabolic enzymes as intervention targets,so as to provide novel ideas for clinical treatment of DKD.

BACKGROUND:Kidney fibrosis represents a key pathological process driving the progression of chronic kidney disease (CKD) and is closely associated with mitochondrial impairment and altered lipid metabolism. Hyperoside, a major flavonoid glycoside from Abelmoschus manihot, has shown anti-fibrotic activity, yet its mechanistic role in renal fibrosis remains unclear. METHODS:Two murine models, folic acid-induced nephropathy and unilateral ureteral obstruction, were employed to assess the renoprotective actions of hyperoside. Mitochondrial function, lipid metabolic remodeling, and fibrotic progression were examined using histological evaluation, biochemical analyses, and ultrastructural assessment by electron microscopy. Integrated transcriptomic and metabolomic analyses were performed to characterize metabolic pathways modulated by hyperoside. Interaction with ACAT1 was confirmed through cellular thermal shift assays, surface plasmon resonance, and molecular docking. Functional relevance was further established through Acat1 knockdown as well as L-carnitine rescue experiments. RESULTS:Hyperoside markedly reduced renal fibrosis and mitochondrial injury in both mouse models. Multi-omics analyses revealed that hyperoside restored fatty acid oxidation and ketone body metabolism. Mechanistically, hyperoside directly bound to and stabilized mitochondrial acetyl-CoA acetyltransferase 1 (ACAT1), suppressing its ubiquitination and enhancing protein stability and enzymatic activity. This interaction promoted l-carnitine-dependent metabolic flux, which activated the NAD⁺-dependent deacetylase SIRT3. SIRT3 in turn deacetylated superoxide dismutase 2 to reduce reactive oxygen species and facilitated mitophagy-mediated clearance of damaged mitochondria. Acat1 knockdown abolished these metabolic and mitochondrial benefits, whereas l-carnitine partially restored them. CONCLUSIONS:Hyperoside mitigates kidney fibrosis by targeting the ACAT1-l-carnitine-SIRT3 axis to reprogram lipid metabolism, improve mitochondrial functional balance, and strengthen antioxidant capacity. Collectively, these results indicate that hyperoside may serve as a potential mitochondria-focused therapeutic strategy for the treatment of CKD.

Introduction: Diabetic kidney disease (DKD) is a devastating complication of diabetes for which there are few potent treatments.Triptolide (TP), an active compound from Tripterygium wilfordii, has shown potential in early studies, but its therapeutic mechanisms in DKD are not fully understood. This study aims to systematically evaluate TP’s efficacy and mechanisms using meta-analysis, network pharmacology, molecular docking, and Mendelian randomization (MR). Methods: A comprehensive search across Chinese and English databases identified animal randomized controlled trials (RCTs) assessing the effects of TP on DKD. A total of 27 studies were incorporated, and a metaanalysis was conducted via Review Manager. TP's drug and disease targets were identified through network pharmacology and molecular docking, while bioinformatics methods were employed to explore the mechanisms. MR analysis was performed to assess potential causal relationships between TP and DKD-related targets. Results: Meta-analysis showed that TP significantly reduced urinary protein, blood lipids, and glucose levels, while improving renal function, renal weight, and renal index (all p < 0.05). Seven core targets—IFNG, CXCL8, TNF, TGFB1, IL2, IL4, and RELA—were identified via network pharmacology, involving key pathways such as lipid-atherosclerosis, AGE-RAGE, and IL-17 signaling. Molecular docking demonstrated strong binding affinities between TP and these targets, with binding energies below -7.00 kJ/mol. Although MR analysis did not establish direct causal relationships between these core genes and DKD, a significant negative correlation between TNF, IL4, and GFR was observed, suggesting their involvement in DKD progression. Discussion: TP may exert therapeutic effects on DKD through coordinated regulation of immune and inflammatory pathways. The integration of multi-omics approaches supports its multi-target pharmacological mechanisms. Although MR analysis did not confirm direct causal relationships, the identified gene associations further reinforce the potential biological relevance of TP. However, this study was primarily based on public datasets and lacks experimental validation in vivo and in vitro. Conclusion: TP exerts therapeutic effects on DKD through multi-target and multi-pathway mechanisms, primarily involving immunomodulation, anti-inflammation, anti-oxidation, and anti-fibrosis processes.

ETHNOPHARMACOLOGICAL RELEVANCE:Kunkui Baoshen Granule (KBG) is a traditional Chinese Medicine (TCM) formula derived from clinical experience and is composed of four medicinal herbs. It has been used for many years at the Jiangsu Province Hospital of Chinese Medicine. It has obtained both a patent and an in-hospital preparation approval from the Jiangsu Provincial Medical Products Administration. KBG has been consistently used to treat diabetic kidney disease (DKD) and has demonstrated significant therapeutic efficacy. AIM OF STUDY:DKD is one of the most prevalent microvascular complications of diabetes mellitus and a leading cause of end-stage renal disease. Mitochondrial dysfunction is closely associated with the development and progression of DKD. KBG has been demonstrated to reduce proteinuria and impede DKD progression. However, further research is required to elucidate the mechanisms by which KBG treats DKD. This study explored the therapeutic effects of KBG in DKD and the underlying molecular mechanisms. METHODS:Blood-circulating compounds from KBG were identified using serum pharmacochemistry. In the in vivo experiments, db/db mice were treated with different dosages of KBG. Biochemical parameters, renal histopathological changes, apoptotic indices, inflammatory factors, and oxidative stress levels were systematically evaluated. Potential targets and mechanisms of action were investigated using proteomic analysis. Molecular docking and molecular dynamics simulations were used to preliminary validate potential therapeutic targets. Untargeted metabolomics was used to analyze the serum metabolic profiles of mice. The expression levels of key proteins were assessed using Western blot, ELISA, immunohistochemistry and immunofluorescence. In vitro experiments, HK-2 cells treated with high glucose and palmitic acid as a cellular model. Inhibitors or lentivirus-mediated knockdown of key targets were employed to further validate the underlying mechanisms. RESULTS:A total of 11 compounds in KBG were identified as prototype components that could enter the bloodstream. In vivo experiments demonstrated that KBG improved renal function, alleviated pathological damage, and mitigated epithelial-mesenchymal transition, inflammation, and oxidative stress in db/db mice. Through proteomics, mitochondrial function and mitophagy were identified as potential mechanisms underlying the therapeutic effects of KBG. Molecular docking and molecular dynamics simulations confirmed good binding stability between the blood-circulating compounds and the key target SIRT3. KBG inhibited NLRP3 inflammasome activation by regulating mitophagy through the SIRT3/FOXO3a signaling pathway. Untargeted metabolomics analysis revealed that KBG primarily remodeled metabolic homeostasis by regulating tryptophan metabolism, TCA cycle, and pyruvate metabolism. Furthermore, in vitro cell experiments demonstrated that KBG improved mitochondrial function and prevented NLRP3 inflammasome activation. Notably, the combination with 3-methyladenine or SIRT3 knockdown abolished the protective effects of KBG in HK-2 cells. CONCLUSION:KBG regulated mitochondrial homeostasis and mitophagy through the SIRT3/FOXO3a signaling pathway, thereby inhibiting NLRP3 inflammasome activation. KBG modulated various metabolic pathways and mitigated metabolic dysregulation. These findings indicate that KBG is a potential multi-component TCM formulation for the effective treatment of DKD.

BACKGROUND:Xietu Hemu prescription (XHP), a Chinese patent formula, is optimized based on the theory of "phlegm-dampness" and has been clinically validated to effectively combat metabolic dysfunction-associated steatotic liver disease (MASLD). It notably reduces visceral fat and body mass index. However, the molecular mechanisms underlying its regulation of lipid metabolism homeostasis remain unexplored. AIM:To elucidate the mechanisms by which XHP inhibits adipocyte differentiation and maintains lipid metabolism homeostasis. METHODS:The therapeutic efficacy of XHP in metabolic-related disorders was analyzed using HepG2 cells and 3T3-L1 cells, along with transcriptomics to assess gene expression alterations during white adipogenesis. The primary metabolites of XHP were identified through ultra-performance liquid chromatography, and metabolic pathways were examined via serum metabolomics. Network analysis was employed to predict therapeutic targets. The accumulation of lipid droplets and the expression of associated proteins were confirmed using oil red O staining and Western blotting, respectively. Molecular docking was utilized to identify core targets and signaling pathways, which were substantiated through immunofluorescence and siRNA interference. RESULTS:XHP-containing serum (XHPS) significantly inhibited the transformation of normal HepG2 cells into fatty liver cells. Concurrently, the treatment suppressed the differentiation of 3T3-L1 cells, reduced lipid droplet accumulation and total cholesterol/triglyceride levels, and downregulated the expression of PPARγ, C/EBPα, and FABP4. Through transcriptomics and network pharmacological intersectionality analyses, 24 core targets were identified, predominantly enriched in the AMPK signaling pathway. Molecular docking validated the strong binding affinity of XHP metabolites to targets such as leptin (-11.3 kcal/mol) and ADIPOQ (-9.4 kcal/mol). ELISA results indicated that XHPS augmented leptin autocrine secretion, thereby activating the AMPK signaling pathway (P < 0.05). Conversely, LEPR knockdown negated this effect (P < 0.05). CONCLUSION:XHP effectively inhibits adipogenesis and enhances lipid metabolism homeostasis through the LEP/AMPK/PPARγ pathway, presenting a promising multi-target therapeutic strategy for MASLD by mitigating lipotoxicity.

PurposeThis study aims to elucidate the mechanistic role of Per- and Polyfluoroalkyl Substances (PFAS) in the pathogenesis and progression of diabetic kidney disease (DKD).MethodsThis study systematically evaluated the toxicity profiles of PFAS compounds utilizing PubChem, ProTox 3.0, and ChEMBL databases. Potential PFAS-related targets were predicted through SwissTargetPrediction and SuperPred platforms. Gene targets associated with DKD were compiled from the GeneCards and OMIM databases. Intersection analysis of PFAS and DKD-related targets was performed to identify candidate genes. A protein-protein interaction network was constructed using STRING to delineate hub targets. Functional enrichment analyses were subsequently conducted via DAVID to elucidate underlying biological processes and pathways. Validation of hub targets encompassed immunohistochemical staining, single-cell expression profiling, subcellular localization assays, and gene expression analyses using external datasets from the Human Protein Atlas (HPA) and Gene Expression Omnibus (GEO). Furthermore, correlations between immune cell infiltration and gene set enrichment analysis (GSEA) were performed to investigate potential mechanistic links. Finally, molecular docking simulations of PFAS compounds with hub proteins were executed using Discovery Studio and CDOCKER to predict binding interactions.ResultsA total of 424 PFAS-associated targets were identified, alongside 9,999 potential toxic targets related to DKD. KEGG pathway enrichment analysis revealed that PFAS toxicity in DKD is implicated in critical signaling pathways, including nitrogen metabolism, peroxisome proliferator-activated receptor (PPAR) signaling, endocrine resistance, insulin resistance, and AMP-activated protein kinase (AMPK) signaling. Hub targets identified comprised MMP9, BCL2, CYP3A43, ACE, HNF4A, HSP90AA1, AGTR1, MMP2, AGTR2, and HMGCR. GSEA further indicated that these hub targets may contribute to immune-mediated renal injury. Molecular docking simulations substantiated strong binding affinities between PFAS compounds and the identified hub proteins, supporting their potential mechanistic involvement.ConclusionThis study provides a theoretical framework for elucidating the toxic targets and underlying mechanisms through which PFAS contribute to the pathogenesis of DKD.

Formononetin exhibits potent anti-oxidative and anti-inflammatory properties, but its precise therapeutic targets and mechanisms in diabetic kidney disease (DKD) remain insufficiently defined. This study evaluated the nephroprotective potential of formononetin using both in vitro (HK-2 cells) and in vivo (db/db mice) DKD models. By integrating network pharmacology and RNA sequencing, the antifibrotic actions of formononetin were further elucidated. Mechanistic investigations revealed that the compound reduced renal fibrosis by suppressing TGF-[Formula: see text]1, FN, and [Formula: see text]-SMA expression, and also alleviated renal dysfunction markers, including UACR, Scr, BUN, 24hUTP, KIM-1, and NGAL. These effects were mediated through the modulation of two key pathways such that the inhibition of the PI3K/AKT/mTOR cascade reduced inflammatory and fibrotic signaling, while the activation of the p38/MAPK axis enhanced autophagic flux, and thus promoted tubular epithelial cell homeostasis. Collectively, these findings support formononetin as a promising candidate for DKD therapy due to its combined anti-inflammatory and pro-autophagic mechanisms.

Extensive evidence suggests that dyslipidemia is associated with endothelial dysfunction, oxidative stress, and inflammation, all of which can contribute to kidney dysfunction. The atherogenic index of plasma (AIP) is a novel marker of lipid metabolism disorder, but its role in kidney dysfunction in diabetic individuals remains controversial. This study aims to clarify the association of AIP with kidney dysfunction in diabetic individuals. This cross-sectional study analyzed a representative sample of participants aged 20 years and older from the United States (n = 2,386, NHANES 2007–2018) and Korea (n = 698, KNHANES 2012). Weighted multivariate logistic regression analyses and smoothed curve fitting were conducted to investigate the relationship between logarithmically transformed AIP (lgAIP) and multiple kidney dysfunction, including albuminuria and low estimated glomerular filtration rate (eGFR) in diabetic individuals. Additionally, we conducted interaction analyses and subgroup analyses to assess whether this relationship remained consistent across different populations. We utilized receiver operating characteristic (ROC) curves to assess and compare the diagnostic performance of AIP and other lipid indices for kidney dysfunction. In both databases, higher lgAIP was significantly associated with the occurrence of albuminuria in diabetic individuals (NHANES: OR = 7.69, 95