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.

NASH poses a significant threat to human health and is recognized as the leading contributor to HCC. In this study, we leveraged publicly accessible datasets to identify novel differentially expressed genes that may serve as potential targets in NASH or potentially NASH-induced HCC. The publicly available datasets were obtained from the GEO. Differential gene expression analysis and enrichment analysis was performed. Subsequently, WGCNA and PPI network were constructed. Lastly, machine learning was employed to identify key feature genes. Utilizing the integrated GEO database, we identified 446 genes exhibiting differential expression. Enrichment analysis indicated that these genes are predominantly associated with glucose and lipid metabolism and inflammatory processes. Through WGCNA, three modules were identified that demonstrated a significant correlation with NASH. Furthermore, core genes among the differentially expressed genes were extracted via protein and protein interaction analysis. Ultimately, machine learning techniques were employed, leading to the identification of three genes: FosB, Fos, and SOCS3. Notably, FosB exhibited consistent expression across various datasets, demonstrated strong predictive capabilities for NASH, and was associated with improved prognostic outcomes in hepatocellular carcinoma by data from TCGA. Additionally, in vitro immunohistochemistry experiments revealed significant reduction of FosB expression in NASH. Bioinformatics analyses conducted on various datasets, along with in vitro immunohistochemistry experiments, revealed significant downregulation of FosB in NASH. It indicates that FosB plays a critical role in the pathogenesis of NASH, and its expression is associated with the prognosis of patients with HCC. Further experimental studies are required to investigate the potential targeting of FosB in NASH and NASH-induced HCC.

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.

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

Background:Given the absence of specific pharmacological treatments for sarcopenia, identifying effective lifestyle and dietary interventions is imperative. This study aims to explore the association between the composite dietary antioxidant index (CDAI) and sarcopenia, offering new insights into nutritional strategies for sarcopenia-prone populations. Methods:This cross-sectional study analysed secondary data from the National Health and Nutrition Examination Survey cycles spanning 2001-2006 and 2011-2018. Weighted multivariate logistic regression and restricted cubic spline (RCS) analyses were employed to evaluate the non-linear association between CDAI and sarcopenia and to perform stratified analyses. Results:In this study, encompassing 19 683 American adults, representative of the national population of 132 140 502 residents, 7.97% were diagnosed with sarcopenia. Across all adjusted models, a higher CDAI was inversely associated with the risk of sarcopenia (OR 0.94, 95% CI 0.92, 0.96; P<0.0001). The highest quartile of CDAI scores to those in the lowest revealed significantly reduced odds of sarcopenia (OR 0.46, 95% CI 0.38, 0.56; P<0.0001). RCS analysis demonstrated a non-linear relationship between CDAI and sarcopenia. Additionally, stratified analyses indicated that the inverse association between CDAI and sarcopenia was more pronounced among participants with higher educational levels and those diagnosed with tumours. Conclusions:There was a negative relationship between CDAI scores and the prevalence of sarcopenia, suggesting that higher CDAI scores may help in managing and preventing the occurrence of sarcopenia.

OBJECTIVES:Immune inflammation plays a crucial role in the pathogenesis of diabetic kidney disease (DKD), but an exact assessment of indicators remains undefined. In this study we address the link between systemic immune-inflammation index (SII) and mortality risk in DKD, and we explore the effect of sex disparities. METHODS:Data from patients with DKD from the National Health and Nutritional Examination Surveys (NHANES, 1999 to 2018) were studied and their causes of death were identified from NHANES-related files. A weighted Cox model was used to evaluate hazard ratios for all-cause, cardiovascular, and cardiocerebrovascular mortality, and these associations were visualized by smoothing curves. RESULTS:The average SII was 634.20 (103/μL). There were 1,283 deaths recorded during 273,422 person-months (396 were cardiovascular related and 461 were cardiocerebrovascular related). Higher SIIs in the fifth quintile were significantly associated with increased mortality (p<0.01). SII trends showed an increased risk of all-cause mortality of >697.0 (103/μL), cardiovascular risk of >717.8 (103/μL), and cardiocerebrovascular risk of >650.0 (103/μL). Mortality increased when SII reached 500 to 660 (103/μL) in men and 700 to 760 (103/μL) in women. CONCLUSIONS:There was a significant association between higher SII and increased risk of all-cause, cardiovascular, and cardiocerebrovascular mortality in DKD patients. In addition, although men had lower SII, their mortality was higher than that of women.

Metabolic dysfunction-associated steatotic liver disease (MASLD) encompasses various conditions, ranging from simple steatosis to metabolic dysfunction-associated steatohepatitis (MASH) and cirrhosis. MASLD is a significant risk factor for hepatocellular carcinoma (HCC) and is rapidly becoming the primary cause of liver transplantation. Dysregulated sphingolipid metabolism has been linked to the development of MASH-HCC. However, detailed insight into the sphingolipid profiles and cell type-specific changes in key genes involved in sphingolipid metabolism remains limited and forms the primary focus of this study. This study used the well-characterized diet-induced MASH-HCC mouse model (DIAMOND). Total RNA sequencing data, NanoString nCounter® Gene profiling, and single-nucleus RNA sequencing (snRNA-seq) GEO data (GSE225381) were used in characterizing gene regulation in MASH-HCC progression. Sphingolipids in the serum and liver were profiled using targeted lipidomics. RNA data analysis showed dysregulation of key genes involved in sphingolipid metabolism, including ceramide synthase 6 (Cers6), serine palmitoyltransferase long chain base subunit 2 (Sptlc2), sphingosine kinase 2 (SphK2), and sphingosine-1-phosphate receptor 1–3 (S1pr1-3) which paralleled significant changes in sphingolipid composition and levels in both serum and liver. Furthermore, TCGA-LIHC patient data were analyzed and potential prognostic genes for MASH-HCC were identified using univariate and multivariate Cox analysis. The multivariate Cox analysis underscored the prognostic significance of several genes related to sphingolipid metabolism, including CERS6, SPTLC2, and S1PR1. Our findings provided valuable insights into the role of sphingolipids in the progression of MASH to HCC. Specific serum and liver sphingolipid profiles may serve as valuable biomarkers for diagnosis and prognosis in MASH-HCC.

BACKGROUND:The duodenum plays a significant role in metabolic regulation, and thickened mucous membranes are associated with insulin resistance. Duodenal mucosal resurfacing (DMR), a new-style endoscopic procedure using hydrothermal energy to ablate this thickened layer, shows promise for enhancing glucose and lipid metabolism in type 2 diabetes (T2D) patients. However, the mechanisms driving these improvements remain largely unexplored. AIM:To investigate the mechanisms by which DMR improves metabolic disorders using a rat model. METHODS:Rats with T2D underwent a revised DMR procedure via a gastric incision using a specialized catheter to abrade the duodenal mucosa. The duodenum was evaluated using histology, immunofluorescence, and western blotting. Serum assays measured glucose, lipid profiles, lipopolysaccharide, and intestinal hormones, while the gut microbiota and metabolomics profiles were analyzed through 16S rRNA gene sequencing and ultra performance liquid chromatography-mass spectrum/mass spectrum, severally. RESULTS:DMR significantly improved glucose and lipid metabolic disorders in T2D rats. It increased the serum levels of cholecystokinin, gastric inhibitory peptide, and glucagon-like peptide 1, and reduced the length and depth of duodenal villi and crypts. DMR also enhanced the intestinal barrier integrity and reduced lipopolysaccharide translocation. Additionally, DMR modified the gut microbiome and metabolome, particularly affecting the Blautia genus. Correlation analysis revealed significant links between the gut microbiota, metabolites, and T2D phenotypes. CONCLUSION:This study illustrates that DMR addresses metabolic dysfunctions in T2D through multifaceted mechanisms, highlighting the potential role of the Blautia genus on T2D pathogenesis and DMR's therapeutic impact.

BACKGROUND:Diabetic kidney disease (DKD) stands as the key contributor to chronic kidney disease worldwide. Clinical studies have shown that Kunkui Baoshen decoction (KKBS) effectively reduces proteinuria and enhances renal function in DKD patients. However, its precise molecular targets and therapeutic mechanisms remain to be thoroughly clarified. AIM:To evaluate the nephroprotective efficacy of KKBS in DKD and explore the underlying mechanisms of action. METHODS:Liquid chromatography-tandem mass spectrometry was utilized to analyze the chemical constituents of KKBS. Metabonomic and transcriptomic analyses were conducted to identify key targets and pathways associated with the therapeutic effects of KKBS on DKD. The nephroprotective effects of KKBS were assessed both in high glucose-induced human kidney-2 cells and in db/db mice. A variety of assays were performed, including Cell Counting Kit-8, Western blot, quantitative reverse transcription-polymerase chain reaction, immunofluorescence, co-immunoprecipitation, periodic acid-Schiff staining, Masson staining, hematoxylin and eosin staining, immunohistochemistry, and mitochondrial morphology analysis. RESULTS:The glutathione metabolic pathway emerged as the most prominent metabolic pathway in the metabonomic analysis of KKBS. Transcriptomic and bioinformatic analyses revealed that nuclear receptor coactivator 4 (NCOA4) was instrumental in regulating ferroptosis within renal tubules of mice with DKD. Both in vitro and in vivo experiments showed that KKBS ameliorated renal dysfunction, mitigated renal tissue damage, and repressed the expression of autophagy-dependent ferroptosis markers and inflammatory fibrosis. Mechanistically, KKBS enhanced the interaction between the homologous to E6-AP C-terminus and RCC1-like domain-containing E3 ubiquitin protein ligase (HERC2) and NCOA4, leading to K48-related ubiquitination and subsequent degradation of NCOA4. This process inhibited autophagy-dependent ferroptosis, reduced the release of pro-fibrotic inflammatory factors, and ultimately exerted an anti-fibrotic effect in DKD. CONCLUSION:KKBS confers nephroprotection in DKD by modulating HERC2/NCOA4-mediated autophagy-dependent ferroptosis, thereby alleviating renal fibrosis.

In addition to histological evaluation for nonalcoholic fatty liver disease (NAFLD), a comprehensive analysis of the metabolic landscape is urgently needed to categorize patients into distinct subgroups for precise treatment. In this study, a total of 806 NAFLD and 267 normal liver samples were comprehensively analyzed. Alterations in 114 metabolic pathways were investigated and two distinct metabolic clusters were identified. Single-cell RNA sequencing (scRNA-seq) analysis was utilized to decipher the metabolic activities within the microenvironment of NAFLD-derived liver cirrhosis. A refined fibrosis prediction model was developed using a Gaussian Mixture Model (GMM), demonstrating superior performance in fibrosis discrimination across multiple independent cohorts. Additionally, using The Cancer Genome Atlas (TCGA), CACNB1 protein was identified as a promising therapeutic target for hepatocellular carcinoma (HCC) patients with elevated metabolic dysfunction scores (MBDS). Machine learning algorithms were applied to MBDS-related genes to select an optimal prognostic model for HCC. All the models were trained in an HCC cohort obtained from the Gene Expression Omnibus (GEO), and the best model was validated in two independent HCC datasets: the TCGA-HCC cohort and LIRI-JP cohort. Overall, we provide insights of metabolic molecular subtyping and its potential clinical applicability in risk stratification for NAFLD and HCC individuals.

Renal fibrosis (RF) is the end stage of several chronic kidney diseases. Its series of changes include excessive accumulation of extracellular matrix, epithelial-mesenchymal transition (EMT) of renal tubular cells, fibroblast activation, immune cell infiltration, and renal cell apoptosis. RF can eventually lead to renal dysfunction or even renal failure. A large body of evidence suggests that natural products in traditional Chinese medicine (TCM) have great potential for treating RF. In this article, we first describe the recent advances in RF treatment by several natural products and clarify their mechanisms of action. They can ameliorate the RF disease phenotype, which includes apoptosis, endoplasmic reticulum stress, and EMT, by affecting relevant signaling pathways and molecular targets, thereby delaying or reversing fibrosis. We also present the roles of nanodrug delivery systems, which have been explored to address the drawback of low oral bioavailability of natural products. This may provide new ideas for using natural products for RF treatment. Finally, we provide new insights into the clinical prospects of herbal natural products.

Background and aims Primary sclerosing cholangitis (PSC) is a chronic liver disease characterized by progressive biliary inflammation and bile duct injury. Berberine (BBR) is a bioactive isoquinoline alkaloid found in various herbs and has multiple beneficial effects on metabolic and inflammatory diseases, including liver diseases. This study aimed to examine the therapeutic effect of BBR on cholestatic liver injury in a PSC mouse model (Mdr2 −/− mice) and elucidate the underlying mechanisms. Methods Mdr2 −/− mice (12–14 weeks old, both sexes) received either BBR (50 mg/kg) or control solution daily for eight weeks via oral gavage. Histological and serum biochemical analyses were used to assess fibrotic liver injury severity. Total RNAseq and pathway analyses were used to identify the potential signaling pathways modulated by BBR in the liver. The expression levels of key genes involved in regulating hepatic fibrosis, bile duct proliferation, inflammation, and bile acid metabolism were validated by qRT-PCR or Western blot analysis. The bile acid composition and levels in the serum, liver, small intestine, and feces and tissue distribution of BBR were measured by LC–MS/MS. Intestinal inflammation and injury were assessed by gene expression profiling and histological analysis. The impact on the gut microbiome was assessed using 16S rRNA gene sequencing. Results BBR treatment significantly ameliorated cholestatic liver injury, evidenced by decreased serum levels of AST, ALT, and ALP, and reduced bile duct proliferation and hepatic fibrosis, as shown by H&E, Picro-Sirius Red, and CK19 IHC staining. RNAseq and qRT-PCR analyses indicated a substantial inhibition of fibrotic and inflammatory gene expression. BBR also mitigated ER stress by downregulating Chop, Atf4 and Xbp-1 expression. In addition, BBR modulated bile acid metabolism by altering key gene expressions in the liver and small intestine, resulting in restored bile acid homeostasis characterized by reduced total bile acids in serum, liver, and small intestine and increased fecal excretion. Furthermore, BBR significantly improved intestinal barrier function and reduced bacterial translocation by modulating the gut microbiota. Conclusion BBR effectively attenuates cholestatic liver injury, suggesting its potential as a therapeutic agent for PSC and other cholestatic liver diseases.

Background Nonalcoholic fatty liver disease (NAFLD) is a leading public health problem worldwide. Approximately one fourth of patients with nonalcoholic fatty liver (NAFL) progress to nonalcoholic steatohepatitis (NASH), an advanced stage of NAFLD. Hence, there is an urgent need to make a better understanding of NAFLD heterogeneity and facilitate personalized management of high-risk NAFLD patients who may benefit from more intensive surveillance and preventive intervene. Methods In this study, a series of bioinformatic methods were performed to identify NAFLD progression-specific pathways and genes, and three machine learning approaches were combined to construct a risk-stratification gene signature to quantify risk assessment. In addition, bulk RNA-seq, single-cell RNA-seq (scRNA-seq) transcriptome profiling data and whole-exome sequencing (WES) data were comprehensively analyzed to reveal the genomic alterations and altered pathways between distinct molecular subtypes. Results Two distinct subtypes of NAFL were identified with the NAFLD progression-specific genes, and one subtype has a high similarity of the inflammatory pattern and fibrotic potential with NASH. The established risk-stratification gene signature could discriminate advanced samples from overall NAFLD. COL1A2, one key gene closely related to NAFLD progression, is specifically expressed in fibroblasts involved in hepatocellular carcinoma (HCC), and significantly correlated with EMT and angiogenesis in pan-cancer. Moreover, the β-catenin/COL1A2 axis might play a critical role in fibrosis severity and inflammatory response during NAFLD-HCC progression. Conclusion In summary, our study provided evidence for the necessity of molecular classification and established a risk-stratification gene signature to quantify risk assessment of NAFLD, aiming to identify different risk subsets and to guide personalized treatment.

Introduction & Objective: Metabolic fatty liver disease (MAFLD) is closely related to dysregulation of systemic metabolism and insulin resistance (IR). Sarcopenia is often observed in individuals with MAFLD. Sphingolipids, known for their pivotal role in muscle cell regeneration and repair, have been suggested as key regulators in this context. This study aims to elucidate the potential connection between sphingolipid dysregulation in muscle and MAFLD. Methods: Ten-week-old male mice with a C57BL/6J and 129S1/SvlmJ mixed background were subjected to either a WDSW (Western diet, Harlan, TD88137, and a high fructose-glucose solution, 23.1g/L D-fructose plus 18.9g/L D-glucose) or standard chow diet for 24 weeks, with free access to food and water. IPGTT and ITT tested were conducted at the end of the study. The sphingolipid profiles in the gastrocnemius muscle were determined using LC-MS/MS. The expression levels of sphingosine-1 phosphate receptors (S1PRs) were measured by qRT-PCR. The GEO data set (GSE73036) was downloaded and analyzed. Result: Mice on the WDSW (MASH mice) exhibited significantly higher blood glucose levels and increased IR after 24 weeks of WDSW feeding. LC-MS/MS analysis indicated elevated levels of C18-Sphingomyelin (SM), C26-SM and C18:1 Dihydroceramide in the muscle of the WDSW group, while S1P levels were markedly reduced. The expression levels of S1PR1-3, the major types of S1PR in muscle, were also significantly decreased in MASH mice. Analysis of GSE73036 highlighted a strong correlation between IR and upregulation of S1P lyase (Spgl), an enzyme crucial for S1P degradation. Conclusion: The findings of this study indicate a clear association between disrupted sphingolipid metabolism and IR in MASH mouse model. The observed increase in ceramide levels coupled with a decrease of S1P may contribute to sarcopenia associated with MAFLD. Disclosure Q. Yan: None. L. Su: None. Y. Tai: None. D. Zhao: None. X. Wang: None. E. Gurley: None. P. Hylemon: None. X. Zhou: None. H. Zhou: None. Funding National Natural Science Foundation Of China (No.82004286)

ObjectivePrevious observational studies have indicated associations between various inflammatory cytokines and diabetic nephropathy (DN) caused by type 2 diabetes mellitus (T2DM). However, the causality remains unclear. We aimed to further evaluate the causal association between 91 inflammatory cytokines and DN using bidirectional two-sample Mendelian randomization (MR) analysis.MethodSummary statistics for DN were obtained from a publicly available genome-wide association study (GWAS) analysis. Data pertaining to inflammatory cytokines were derived from a GWAS protein quantitative trait locus (pQTL) study. The primary analytical approach employed the inverse variance weighted (IVW) method, complemented by MR-Egger regression, weighted mode (WM), and weighted median (WME) methods to evaluate the causal association between inflammatory cytokines and DN. Sensitivity analyses were conducted to validate the robustness of the findings.ResultAmong individuals of European ancestry, the IVW method results revealed a positive causal association between the gene expression of tumor necrosis factor ligand superfamily member 14 (TNFSF14), and TNF-related activation-induced cytokine (TRANCE) with DN. Conversely, a negative causal association was observed between the gene expression of interleukin-1-alpha (IL-1 alpha), and transforming growth factor-alpha (TGF-alpha) with DN. Among individuals of East Asian ancestry, the IVW method results indicated a negative causal association between the gene expression of glial cell line-derived neurotrophic factor (GDNF) and DN. Notably, these findings persisted without evidence of horizontal pleiotropy or heterogeneity, ensuring their robustness and reliability.ConclusionThe MR analysis underscores a causal association between inflammatory cytokines and DN, providing an important reference and evidence for the study of DN.

Background: Diabetic kidney disease (DKD) is one of the most serious microvascular complications of diabetes, with the incidence rate increasing yearly, which is the leading cause of chronic kidney disease (CKD) and end-stage kidney disease. Abelmoschus Manihot capsule, as a proprietary Chinese patent medicine, is widely used for treating CKD in China. Currently, the combination of Abelmoschus Manihot (AM) capsule and renin-angiotensin-aldosterone system inhibitor (RASI) has gained popularity as a treatment option for DKD, with more and more randomized control trials (RCTs) in progress. However, the high-quality clinical evidence supporting its application in DKD is still insufficient.Aim of the study: To comprehensively and systematically evaluate the efficacy and safety of AM capsule combined with RASI in the treatment of DKD.Materials and methods: English and Chinese databases such as Pubmed, Cochrane Library, Embase, CNKI, SinoMed, WF, and VIP were searched to collect the RCTs of AM capsule in treatment of DKD. Then Two investigators independently reviewed and extracted data from the RCTs which met the inclusion criteria. The quality of the data was assessed using the Cochrane risk of bias assessment tool, and meta-analysis was performed using RevMan 5.4 software.Results: 32 RCTs with a total of 2,881 DKD patients (1,442 in the treatment group and 1,439 in the control group) were included. The study results showed that AM capsule combined with RASI could be more effective in decreasing 24h-UTP [MD = −442.05, 95% CI (−609.72, −274.38), p < 0.00001], UAER [MD = −30.53, 95% CI (−39.10, −21.96), p < 0.00001], UACR [MD = −157.93, 95% CI (−288.60, −27.25), p < 0.00001], Scr [MD = −6.80, 95% CI (−9.85, −3.74), p < 0.0001], and BUN [MD = −0.59, 95% CI (−1.07, −0.12), p = 0.01], compared to using RASI alone. According to the subgroup analyses, the combination of AM and ARB seems to be more effective in reducing UAER than the combination of ACEI, and the addition of AM may achieve a more significant clinical effect on decreasing Scr for DKD patients with 24h-UTP>2 g or Scr>110–133 μmol/L and >133 μmol/L. Furthermore, no additional adverse reactions were observed in the combination group [OR = 1.06; 95%CI: (0.66, 1.69), p = 0.82].Conclusion: Combining AM with RASI may be a superior strategy for DKD treatment compared to RASI monotherapy. However, due to significant heterogeneity, the results should be interpreted with great caution, and more high-quality RCTs with multi-centers, different stages of DKD, large sample sizes, and long follow-up periods are still needed to improve the evidence quality of AM for DKD in the future.Systematic Review Registration:https://www.crd.york.ac.uk/PROSPERO/#recordDetails; Identifier CRD42022351422