INTRODUCTION:Metabolic dysfunction-associated steatohepatitis (MASH) represents a severe subtype of metabolic dysfunction-associated fatty liver disease (MAFLD), characterized by hepatic steatosis, chronic inflammation, with or without fibrosis. Without appropriate intervention, MASH can progress to hepatocellular carcinoma (HCC). Lysophosphatidylcholine acyltransferase 3 (LPCAT3), an endoplasmic reticulum membrane protein regulating phospholipid composition, is critical for maintaining endoplasmic reticulum and mitochondrial homeostasis. However, its role in MASH-to-HCC progression remains unclear. OBJECTIVES:To investigate the mechanism by which LPCAT3 influences the progression from MASH to HCC. METHODS:LPCAT3 expression levels were examined in tumor and adjacent non-tumor tissues from patients with MASH-associated HCC. A MASH-HCC mouse model was established using a high-fat, high-cholesterol diet, and lipidomics and proteomics analyses were performed to elucidate the role of LPCAT3 in MASH-HCC progression. In addition, a liver-specific LPCAT3 knockout mouse model was generated, and LPCAT3 was overexpressed in mouse livers using an adeno-associated virus (AAV) system to evaluate its role in the development of MASH-HCC. RESULTS:Our study revealed that hepatic LPCAT3 expression was markedly reduced in mice fed a high-fat, high-cholesterol diet. Liver-specific LPCAT3 deficiency accelerates MASH progression, as evidenced by increased hepatic inflammation and fibrosis, and further promoted the transition from MASH to HCC over time. Mechanistically, LPCAT3 deficiency upregulated protein disulfide isomerase (Pdi)-endoplasmic reticulum oxidoreductase 1 alpha (Ero1α) expression, leading to mitochondrial accumulation of H2O2 and Ca2+ and impairing mitochondrial oxidative phosphorylation function. LPCAT3 deficiency also reduced PC (18:2/18:2) levels, whereas supplementing PC (18:2/18:2) in LPCAT3-knockdown cells reversed the upregulation of Pdi-Ero1α and alleviated mitochondrial H2O2 and Ca2+ accumulation. Furthermore, LPCAT3 overexpression ameliorated mitochondrial dysfunction and inhibited the progression from MASH to HCC in mice. CONCLUSION:LPCAT3 deficiency triggers Pdi-Ero1α-mediated mitochondrial dysfunction, identifying LPCAT3 as a promising therapeutic target for MASH-associated HCC.

AIMS:Metabolic dysfunction-associated steatohepatitis (MASH) is a primary driver of hepatocellular carcinoma (HCC), yet effective therapeutic interventions remain limited. While luteolin is known for its anti-inflammatory properties, its efficacy and underlying mechanism in the MASH-HCC transition are not fully understood. This study investigated the protective effects of luteolin against MASH-HCC and the role of the AMPK/ACC signaling pathway in this process. MATERIALS AND METHODS:In vivo, a MASH-HCC mouse model was established using diethylnitrosamine (DEN) combined with a high-fat, high-cholesterol (HFHC) diet. Mice were treated with vehicle or luteolin (50 or 100 mg/kg) for 26 weeks. Progression was monitored via serum alpha-fetoprotein (AFP), histological analysis, and Western blotting. In vitro, HepG2 and Huh-7 cells were challenged with cholesterol and treated with luteolin. The AMPK inhibitor BAY-3827 was employed to verify whether the metabolic benefits of luteolin were pathway-dependent. KEY FINDINGS:Luteolin treatment significantly reduced tumor burden, lowered serum AFP levels, and attenuated hepatic lipid accumulation and fibrosis in MASH-HCC mice. In vitro results mirrored these findings, showing that luteolin reduced cholesterol-induced lipid loading. Mechanistically, luteolin increased the phosphorylation of AMPK and its downstream target, ACC. Furthermore, pharmacological inhibition of AMPK with BAY-3827 abolished the lipid-lowering effects of luteolin in hepatic cells, confirming that its therapeutic benefits are mediated through AMPK activation. SIGNIFICANCE:Luteolin suppresses the progression of MASH to HCC by activating the AMPK/ACC signaling pathway and subsequently inhibiting de novo lipogenesis. These findings highlight luteolin as a promising potential therapeutic candidate for the prevention and treatment of MASH-related liver cancer.

OBJECTIVE:To develop a protein risk score (ProRS) for predicting liver-related events (LREs) in patients with diabetes and compare its predictive performance with the Fibrosis-4 Index (FIB-4) and an established polygenic risk score. RESEARCH DESIGN AND METHODS:This prospective cohort study included 13 516 individuals with prediabetes and type 2 diabetes (T2D) from the UK Biobank. Cox proportional hazards models and LASSO regression were applied to identify proteins associated with incident LREs and construct the ProRS. Predictive performance was assessed using Harrell's C-index, time-dependent area under the receiver operating characteristic curve, net reclassification improvement and integrated discrimination improvement. RESULTS:Over a median follow-up of 13.5 years, 171 (1.3%) incident LREs occurred. We identified 877 proteins associated with LRE risk, primarily enriched in inflammatory signalling, extracellular matrix remodelling and complement/coagulation cascades. In the training set, we developed a 24-protein ProRS (C-index, 0.842; 95% CI 0.797-0.884) that stratified individuals into low-, medium- and high-risk groups, with 10-year cumulative incidences of LREs of 0.2%, 1.2% and 14.2%, respectively. Compared with the low-risk group, the hazard ratio for LREs was 57.1 (95% CI 31.9-102) in the high-risk group. In the internal validation set, the ProRS model (C-index, 0.876; 95% CI 0.827-0.920) accurately predicted both short- and long-term LREs and outperformed FIB-4 index (C-index, 0.733; 95% CI 0.657-0.807) and polygenic risk score (C-index, 0.636; 95% CI 0.564-0.706). CONCLUSIONS:The protein risk score demonstrated superior performance compared with the FIB-4 index and the polygenic risk score in predicting incident LREs among individuals with prediabetes and T2D. The score allows stratification of individuals according to liver-related risk, though external validation in multi-ethnic cohorts is warranted.

Chronic gastrointestinal pain (CGP) is a common and often difficult-to-manage symptom in disorders of gut-brain interaction (DGBI). Owing to the limited efficacy of current therapeutic approaches in a subset of patients, a better understanding of gut-brain axis (GBA) dysfunction may facilitate the development of improved treatment strategies. This review summarizes the pathophysiological mechanisms underlying CGP, focusing on the contributions of microbial dysbiosis, mucosal immune activation, and neuroendocrine disturbances to peripheral and central nociceptive sensitization. Current therapeutic approaches, including cognitive and behavioral interventions, pharmacological neuromodulation, and microbiome-directed therapies, are critically reviewed with regard to their mechanistic basis and available clinical evidence. In addition, we discuss evidence indicating that plant-derived bioactive compounds have been reported to modulate multiple pathways implicated in CGP, including epithelial barrier dysfunction, visceral hypersensitivity, inflammatory signaling, oxidative stress, and ion channel activity. By integrating advances in psychogastroenterology with emerging findings from natural product research, this review discusses potential complementary strategies for CGP management and the challenges associated with their clinical translation. A deeper understanding of GBA regulation may facilitate the identification of novel therapeutic targets and inform the development of more individualized treatment strategies for CGP, although further preclinical and clinical studies are required to establish their efficacy and safety.

Ceramide is an influential lipid signaling molecule and a key factor in sphingolipid (SL) metabolism; ceramide plays integral roles in the regulation of body metabolism and tumor pathogenesis. Changes in the levels of ceramides and associated enzymes can contribute to metabolic changes and thus cause colorectal cancer (CRC) to enter different survival stages. Elucidating the mechanisms underlying the functions of ceramides and related enzymes in CRC may reveal new therapeutic strategies. In this review, we summarize some of the fundamental mechanisms mediating the effects of ceramide and related enzymes in modulating CRC development via the ceramide-mediated induction of tumor apoptosis, autophagy, proliferation and migration as well as the effects of the key enzymes that are involved in ceramide synthesis and catabolism on the fate of CRC. The aim of this review is to provide novel ideas for developing therapeutic strategies for CRC from a metabolic perspective.

Colorectal cancer (CRC) ranks among the most prevalent malignant neoplasms globally. A growing body of evidence underscores the pivotal roles of genetic alterations and dysregulated epigenetic modifications in the pathogenesis of CRC. In recent years, the reprogramming of tumor cell metabolism has been increasingly acknowledged as a hallmark of cancer. Substantial evidence suggests a crosstalk between tumor cell metabolic reprogramming and epigenetic modifications, highlighting a complex interplay between metabolism and the epigenetic genome that warrants further investigation. Biomarkers associated with the pathogenesis and metabolic characteristics of CRC hold significant clinical implications. Nevertheless, elucidating the genetic, epigenetic, and metabolic landscapes of CRC continues to pose considerable challenges. Here, we attempt to summarize the key genes driving the onset and progression of CRC and the related epigenetic regulators, clarify the roles of gene expression and signaling pathways in tumor metabolism regulation, and explore the potential crosstalk between epigenetic events and tumor metabolic reprogramming, providing a comprehensive mechanistic explanation for the malignant progression of CRC. Finally, by integrating reliable targets from genetics, epigenetics, and metabolic processes that hold promise for translation into clinical practice, we aim to offer more strategies to overcome the bottlenecks in CRC treatment.

Immune checkpoint inhibitors (ICIs) have shown limited efficacy in colorectal cancer (CRC). Chinese yam polysaccharide (CYP), a naturally derived plant polysaccharide, demonstrates immunomodulatory and antitumour activities. This study investigated whether CYP enhances the antitumour effects of αPD-1 monoclonal antibody (mAb) by modulating gut microbiota and metabolites. In MC38 and CT26 xenograft models, CYP synergistically inhibited tumour growth when combined with αPD-1 mAb. 16S rRNA sequencing revealed that the combination therapy enriched beneficial bacteria (such as Clostridia_UCG-014 and Actinobacteria) while reducing pathogenic bacteria (including Enterorhabdus and Desulfovibrionaceae). Antibiotic-mediated gut microbiota ablation abolished therapeutic benefits, confirming microbiota-dependent mechanisms. Cytometry by Time-Of-Flight indicated that the combination therapy reshaped the tumour microenvironment by inhibiting immunosuppressive M2 macrophages (CD206+ subset) and enhancing infiltration of cytotoxic CD8+ T cells. Metabolomics analysis demonstrated that the combination therapy effectively rectified tumour-induced metabolic dysregulation, particularly in pathways related to linoleic acid, tryptophan, and purine metabolism. Significantly, the purine-associated metabolite deoxyguanosine was identified to promote M2 macrophage polarization and tumour progression in vitro, whereas its levels were markedly attenuated following combined therapeutic intervention. The results suggest that CYP enhances the efficacy of αPD-1 mAb through remodeling gut microbiota, reducing pro-tumour metabolite (deoxyguanosine), and reprogramming the tumour immune microenvironment. This provides a novel strategy for enhancing CRC patients' response to anti-PD-1 immunotherapy response.

Background Uric acid (UA) is the terminal product of purine metabolism. Elevated serum uric acid (SUA) levels, resulting from excessive synthesis or impaired excretion, are link to chronic inflammatory stress and increased risks of colorectal, breast, and prostate cancers. Hyperuricemia triggers a cascade of proinflammatory and oxidative responses, establishing a microenvironment conducive to tumorigenesis. Aim of review This review synthesizes evidence on how hyperuricemia drive inflammation and cancer transformation from global foundational research and clinical practice, elucidate UA metabolism as potential therapeutic strategy for inflammation-associated malignancies. Key scientific concepts of review Hyperuricemia-induced oxidative stress, DNA damage and genomic instability, while simultaneously activating proinflammatory signaling pathways. These interconnected pathways establish a persistent, proinflammatory microenvironment that fosters the transition from inflammation to cancer. Therapeutic strategies targeting UA metabolism (including pharmacologic interventions and dietary modifications) may mitigate chronic low-grade inflammation and reduce the cancer risk associated with hyperuricemia. Dysregulated UA metabolism emerges as a critical modulator linking chronic inflammation with oncogenesis.

RNA methylation modifications, as a widespread type of modification in eukaryotic cells, especially N6-methyladenosine (m6A), are associated with many activities in organisms, including macrophage polarization and progression of non-alcoholic steatohepatitis (NASH). Macrophages in the liver are of diverse origin and complex phenotype, exhibiting different functions in development of NASH. In the review, we discuss the functions of m6A and m6A-related enzymes in macrophage polarization. Furthermore, we retrospect the role of macrophage polarization in NASH. Finally, we discuss the prospects of m6A in macrophages and NASH, and provide guidance for the treatment of NASH.

Ethnopharmacological relevance: Liansu capsule could alleviate dyspeptic symptoms; however, the mechanisms underlying its role in treating functional dyspepsia (FD) remain unclear. Aim of the study: To elucidate the mechanism underlying the efficacy of Liansu capsule in alleviating FD symptoms. Materials and methods: Thirty-six male mice were randomly divided into the following six groups: control, model, low-strength Liansu, moderate-strength Liansu, high-strength Liansu, and domperidone groups. Small intestine propulsion rate, gastric residual rate and histopathological analysis were performed to evaluate efficacy of Liansu capsule. Levels of interleukin-1 beta, interleukin-6, tumor necrosis factor alpha, phosphorylation of p65, ghrelin and gastrin were verified by real-time quantitative polymerase chain reaction and immunofluorescence assays. Targeted metabolomic analyses, western blotting and immunofluorescence assays were used to explore the mechanism of Liansu capsule in ameliorating FD. Results: The Liansu capsule significantly ameliorated the symptoms of FD, and markedly increased the levels of ghrelin and gastrin. Moreover, Liansu capsule significantly downregulated the levels of the proinflammatory cytokine interleukin-1 beta, interleukin-6, tumor necrosis factor alpha, and inhibited the phosphorylation of p65. Targeted metabolomic analyses showed that Liansu capsule significantly reduced the levels of deoxycholic acid and hyodeoxycholic acid, which were significantly elevated in the model group. Furthermore, these results showed that deoxycholic acid and hyodeoxycholic acid markedly promoted the levels of Takeda G-protein-coupled receptor 5 (TGR5), phosphorylated signal transducer and activator of transcription 3 (STAT3), and Kruppel-like factor 5 (KLF5) in vitro. whereas, Liansu capsule significantly reduced the levels of TGR5, phosphorylated STAT3, and KLF5. Conclusion: Our findings indicated that Liansu capsule improved FD by regulating the deoxycholic acid/hyodeoxycholic acid-TGR5-STAT3-KLF5 axis. The findings reveal a novel mechanism underlying the role of Liansu capsule, which may be a promising therapeutic strategy for FD.

Introduction:Gan–jiang–ling–zhu (GJLZ) decoction is a classical traditional Chinese medicine prescription. Through invigorating yang, activating qi and dissipating dampness, GJLZ decoction is widely applied for the treatment of chronic digestive disease, including nonalcoholic fatty liver disease. However, efficacy and mechanism of GJLZ decoction behind nonalcoholic steatohepatitis (NASH) treatment remains unelucidated.Methods: NASH was induced in mice, followed by treatment with GJLZ decoction. Various methods including hematoxylin-eosin, oil red O staining, and triglyceride analysis were employed to evaluate the treatment effects of GJLZ decoction on NASH. Gut microbiota, metabolomics, cell viability assays, immunofluorescence and Western blotting were performed to unveil the mechanism behind GJLZ decoction.Results: GJLZ decoction treatment significantly improved hepatic steatosis in mice with NASH. It led to remodeling of gut flora and metabolite structures, including the 12-tridecenoic acid level. 12-Tridecenoic acid aggravated hepatic steatosis by promoting acetyl-coenzyme A carboxylase alpha (ACC) expression and inhibiting carnitine palmitoyltransferase 1A (CPT1A) expression. GJLZ decoction treatment reduced the 12-tridecenoic acid level, inhibited ACC activity and promoted CPT1A expression.Conclusion: Our results demonstrated that 12-tridecenoic acid aggravated hepatic steatosis by affecting the ACC–CPT1A axis and GJLZ decoction treatment effectively reduced the 12-tridecenoic acid level and improved steatosis.

Ethnopharmacological relevance Gan-Jiang-Ling-Zhu (GJLZ) decoction, a classical Chinese herbal prescription, can be applied for the treatment of metabolic diseases including liver steatosis. Although GJLZ decoction has been widely applied clinically for thousands of years, the mechanism of GJLZ decoction behind treatment of nonalcoholic steatohepatitis (NASH) remains relatively unelucidated. Aim of the study To elucidate the efficacy of GJLZ decoction in the treatment of NASH and to investigate its underlying mechanisms from an epigenetic perspective. Materials and methods The quality control of chemical components in GJLZ decoction was conducted. C57BL/6J mice with NASH were induced by feeding them a choline-deficient-high-fat-diet (CDHFD), along with GJLZ decoction intervention for 4 weeks. Then NASH phenotypes including histological steatosis, inflammation, hepatic apoptosis, fibrosis, serum liver enzyme and lipid level were measured. N6-methyladenosine (m6A) and transcriptome sequencing were performed. Levels and functions of methyltransferases and different genes were performed by quantitative polymerase chain reaction, immunofluorescence, gene knockdown, oil red O staining and western blotting. Results GJLZ decoction significantly reduced liver weight, liver index and improved hepatic steatosis, and inflammation, as well as inhibited hepatic apoptosis and fibrosis. Moreover, GJLZ decoction significantly reduced the levels of lactate dehydrogenase, aminotransferase, triglyceride, aspartate aminotransferase, and inhibited levels of interleukin 6 and tumor necrosis factor α. Transcriptome and m6A sequencing revealed the landscape of transcriptome and m6A modification influenced by NASH and the following GJLZ decoction intervention. Eleven differential genes were identified, and GJLZ markedly promoted m6A level of UDP glucuronosyltransferase family 2 member A3 (Ugt2a3), to promote its expression. Additionally, GJLZ significantly promoted methyltransferase 14 (METTL14) expression, whereas METTL14 knockdown aggravated hepatocellular steatosis. Finally, METTL14 knockdown significantly reduced the level of Ugt2a3 by promoting its degradation, whereas, Ugt2a3 overexpression could markedly inhibit hepatocellular steatosis. Conclusions GJLZ decoction demonstrates potential in alleviating CDHFD-induced NASH by modulating the METTL14-m6A-Ugt2a3 axis, offering a novel therapeutic approach for NASH treatment.

Globally, colorectal carcinoma (CRC) ranks third in terms of prevalence according to the latest Global Cancer Statistics.1 Studies have shown that most CRCs begin as preexisting adenomas,2 among which advanced adenomas (AAs) have been demonstrated to be a more intense risk factor.3 Exploring the underlying mechanism of AA-CRC transformation is helpful in providing a basis for the precise treatment of CRC. DNA 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) could play major roles in CRC.4, 5 A single-tube methylation-specific quantitative polymerase chain reaction (PCR) assay could be a good predictor of CRC recurrence,4 and 5-hmC levels of zw10 kinetochore protein could have a high diagnostic performance for early-stage CRC.5 However, the functions of 5mC and 5hmC in AA-CRC transformation remain unclear. Therefore, we conducted an integrated analysis of 5mC and 5hmC to elucidate the mechanism underlying AA-CRC transformation. Detailed information regarding the study design, participant recruitment and methods was provided in Additional File 1. First, to verify the role of 5mC in AA-CRC transformation, 5mC profiles were obtained. Differentially methylated sites (DMSs) and differentially methylated genes (DMGs) were identified (Figure 1A; Figure S1A–C; and Additional File 2). Subsequently, the main biological functions of DMGs were verified, including ubiquitin-mediated proteolysis, the transforming growth factor-beta pathway, and pluripotency of stem cells (Figure S1D,E; Additional File 3). Based on the characterization of 5mC in AA-CRC transformation, the 5mC levels in AA and CRC were further investigated. The results showed that 5mC levels were significantly decreased in AA and then markedly increased in CRC, consistent with the results of sequencing (Figure 1B). Tissue microarrays (TMAs) showed that 5mC levels were also markedly increased in CRC (Figure 1C,D), and patients with CRC with high 5mC levels had a short overall survival (Figure 1E). In addition, 5hmC levels were markedly decreased in CRC tissues (Figure 1F,G). However, no correlation was found between 5hmC levels and the overall survival of patients with CRC (Figure 1H). The levels of 5mC and 5hmC were further verified using immunofluorescence (Figure 1I). These results revealed dynamic changes in 5mC and 5hmC during AA-CRC transformation. 5hmC is a stable derivative catalyzed by tet methylcytosine dioxygenases (TETs) in DNA demethylation. To determine dynamic changes in 5mC and 5hmC, we further analyzed 5mC profiles combined with data of 5hmC published in a previous study.5 Compared with theAA group, the majority of DMSs in 5mC were hypermethylated in the CRC group (Figure 1A), but the majority of DMSs in 5mC+5hmC were hypermethylated (Figure 2A). Hypermethylated changes in 5mC mainly occurred in the open sea, and changes in 5mC+5hmC occurred in the open sea, N-shore, S-shore, N-shelf, S-shelf, and CpG island (Figure 2B). Hypermethylated 5mC DMSs were enriched in other and first exon regions (Figure 2C), while hypermethylated changes in 5mC+5hmC were enriched in other, the transcriptional start site 1500 and first exon regions (Figure 2C). In addition, enrichment of hypermethylated changes in 5mC+5hmC in the enhancer was also found (Figure 2D). We then performed a combinational analysis of hypermethylated genes and hypohydroxymethylated genes, and 20 overlapped genes were chosen (Figure 3A). The levels of five genes (ANO10, SUCLG2, PPARGC1A, LRBA, and ATP8A1) were positively correlated with the overall survival of patients with CRC (Figure S2). Moreover, compared with the AA group, mRNA and protein levels of PPARGC1A, LRBA, and ATP8A1 but not ANO10 and SUCLG2 were both markedly decreased in the CRC group (Figure 3B–F). The 5hmC levels of PPARGC1A, LRBA, and ATP8A1 were markedly decreased, and the levels of 5mC were significantly higher in the CRC group than in the AA group (Figure 3G–I). Analysis of the SurvivalMeth database showed that the 5mC levels of PPARGC1A, LRBA, and ATP8A1 were negatively correlated with the overall survival of patients with CRC (Figure 3J–L). To elucidate the mechanism of AA-CRC transformation, the levels of DNA methylases and demethylases were measured. The results showed that DNMT3B levels were significantly increased in AA-CRC transformation but not DNMT1 and DNMT3A (Figure 4A,B; Figure S3A–C). TET2 levels were significantly decreased in AA-CRC transformation but not TET1 and TET3 (Figure 4C,D; Figure S3D–F). Studies have indicated that DNMT3B can accelerate the invasion and migration of CRC and promote CRC development,6 and TET2 can inhibit CRC progression.7 Our results showed that DNMT3B knockdown and TET2 overexpression significantly inhibited cell proliferation, invasion, and migration (Figure 4E–H; Figure S3G–I). Moreover, DNMT3B knockdown increased the mRNA and protein levels of PPARGC1A and LRBA but not ATP8A1 (Figure 4I; Figure S4A). TET2 overexpression also promoted the protein level of PPARGC1A but reduced LRBA and ATP8A1 protein levels (Figure 4J). Therefore, PPARGC1A may be a downstream target of DNMT3B and TET2. PPARGC1A levels in CRC were negatively associated with DNMT3B levels and positively associated with TET2 levels (Figure S4B–E). Studies have shown that PPARGC1A mediates mitochondrial biogenesis and energy metabolism to regulate tumourigenesis in CRC.8, 9 Our results also showed that PPARGC1A was markedly decreased in CRC, and PPARGC1A overexpression inhibited cell proliferation, invasion, and migration in HCT116 cells (Figure 4K,L; Figure S4F–H). Moreover, PPARGC1A expression was positively correlated with activated dendritic cells, memory resting CD4 T cells, and also related to energy metabolism and mitochondrial gene expression (Figure S4I–K; Additional File 5). In addition, compared with the AA group, the 5-mC level of PPARGC1A was markedly increased and 5-hmC level of PPARGC1A was markedly decreased in the CRC group (Figure 4 M,N). These results indicated that PPARGC1A mediated by DNMT3B and TET2 could regulate AA-CRC transformation.10 In summary, 5mC and 5hmC showed dynamic changes in the progression of AA-CRC transformation. Mechanistically, DNMT3B knockdown and TET2 overexpression inhibited CRC progression. Finally, DNMT3B-mediated 5mC and TET2-mediated 5hmC regulated PPARGC1A expression, which could regulate the progression of AA-CRC transformation (Figure 4O). Our results not only suggest critical roles of DNMT3B and TET2 in the AA-CRC transformation but also provide a new strategy for CRC treatment. The authors would like to thank Cloud-Seq Biotech Inc. (Shanghai, China) for MeDIP-seq and NewCore Biotech (Shanghai, China) for bioinformatics analysis of the data. This work was supported by the Shanghai Rising-Star Program (grant number: 21QA1409000) and Shanghai Frontier Research Base of Disease and Syndrome Biology of Inflammatory Cancer Transformation (grant number: 2021KJ03–12). The authors declare no competing interests. The Shanghai Rising-Star Program, Grant Number: 21QA1409000; Shanghai Frontier Research Base of Disease and Syndrome Biology of Inflammatory Cancer Transformation, Grant Number: 2021KJ03–12 Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

Background:In lean individuals, nonalcoholic fatty liver disease (NAFLD) is not a benign disease, and these patients have long-term morbidity and mortality similar to those of their nonlean counterparts. Finding biomarkers for noninvasive and early detection is urgent and microRNAs (miRNAs) show potential. The aims of this study were to investigate the potential role of serum miRNAs in the detection of lean NAFLD and to explore the possible pathogenesis of lean NAFLD.Methods:A total of 498 patients with NAFLD and 98 healthy controls were included to compare the clinical characteristics of lean NAFLD patients [LNs: body mass index (BMI) <23 kg/m2], nonlean NAFLD patients (NLNs: BMI ≥23 kg/m2) and normal healthy individuals (HIs). A total of 14 serum samples were collected from 4 LNs, 6 NLNs and 4 HIs for high-throughput profiling to identify altered miRNA expression patterns in lean NAFLD. The candidate miRNA, miR-4488, was identified by filtering based on studies in a second independent cohort (31 LNs, 62 NLNs, 72 HIs) that included quantitative real-time polymerase chain reaction (qRT-PCR) analysis. Gene Ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, and protein-protein interaction network analyses were performed to investigate the potential molecular mechanism of miR-4488 in lean NAFLD.Results:LNs were older and had a smaller waist circumference, lower levels of alanine aminotransferase, glutamyl transpeptidase, fasting insulin, and uric acid, lower HOMA-IR score, and higher levels of total cholesterol, high-density lipoprotein cholesterol, and hemoglobin (P<0.05). The serum level of miR-4488 was increased in LNs compared with HIs (P<0.0001) and NLNs (P=0.025). miR-4488 had acceptable performance in predicting [area under the curve (AUC) =0.794, 0.698] lean NAFLD. Moreover, GO and KEGG enrichment analyses revealed that the differentially expressed target genes were mainly involved in choline metabolism in cancer, the tumor-necrosis factor (TNF) signaling pathway and the p53 signaling pathway. PPI analysis identified ARHGAP1, SLC10A1 and SIX5 as the hub genes.Conclusions:Taken together, our findings indicate that serum miR-4488 is a potential biomarker for diagnosing and predicting the pathogenetic mechanisms of lean NAFLD.

BACKGROUND:As a primarily N6-methyladenosine methyltransferase, methyltransferase 3 (METTL3) plays a crucial role in nonalcoholic fatty liver disease. However, its regulatory mechanism in steatosis remains unknown.METHODS:Alpha mouse liver 12 (AML12) cells were induced by free fatty acids (FFA). Triglycerides, lipid droplet assay, and Oil Red O staining were performed to evaluate steatosis. The expression of METTL3 and cytochrome P450 family 4 subfamily f polypeptide 40 (CYP4F40) was measured using Western blotting, real-time quantitative polymerase chain reaction, and dual-luciferase reporter assay. Triglycerides, total cholesterol, almandine aminotransferase, and aspartate aminotransferase were assayed after cinnamaldehyde treatment. Transcriptomics and metabolomics were performed to determine how METTL3 and cinnamaldehyde regulate steatosis.RESULTS:METTL3 protein level was reduced in FFA-induced steatosis in AML12 cells, and METTL3 knockdown aggravated the steatosis. Cinnamaldehyde alleviated steatosis by increasing METTL3 expression. A combined transcriptomics and metabolomics analysis revealed that METTL3 knockdown reduced CYP4F40 expression and reduced the level of capric acid, gamma-linolenic acid, arachidonic acid, and docosapentaenoic acid. Cinnamaldehyde promoted CYP4F40 expression by increasing METTL3 and increased the levels of capric acid, gamma-linolenic acid, arachidonic acid, and docosapentaenoic acid. Finally, the beneficial effects of cinnamaldehyde on steatosis were reversed after METTL3 knockdown.CONCLUSIONS:METTL3 knockdown aggravated steatosis in AML12 cells through CYP4F40-mediated fatty acid metabolism, and cinnamaldehyde alleviated steatosis via the METTL3-CYP4F40 pathway.

As the predominant type of chronic liver disease, the growing prevalence of nonalcoholic fatty liver disease (NAFLD) has become a concern worldwide. Although obesity plays the most pivotal role in NAFLD, approximately 10-20% of individuals with NAFLD who are not overweight or obese (BMI < 25 kg/m2, or BMI < 23 kg/m2 in Asians) have "lean NAFLD." Lean individuals with NAFLD have a lower prevalence of diabetes, hypertension, hypertriglyceridemia, central obesity, and metabolic syndrome than nonlean individuals with NAFLD, but higher fibrosis scores and rates of cardiovascular morbidity and all-cause mortality in advanced stages. The pathophysiological mechanisms of lean NAFLD remain poorly understood. Studies have shown that lean NAFLD is more correlated with factors such as environmental, genetic susceptibility, and epigenetic regulation. This review will examine the way in which the research progress and characteristic of lean NAFLD, and explore the function of epigenetic modification to provide the basis for the clinical treatment and diagnosis of lean NAFLD.

Nonalcoholic fatty liver disease (NAFLD) is a general term for a series of liver diseases including simple steatosis, non-alcoholic steatohepatitis, liver fibrosis, which is closely related to metabolic syndrome. The pathogenesis of NAFLD is relatively complex, which has gradually changed from the previous 'two-hit' hypothesis to the current "multiple hits" hypothesis. However, there is currently no approved treatment for NAFLD in clinic, highlighting the urgent need for drug development. Peroxisome proliferator activated receptors (PPARs) are members of the nuclear receptor superfamily, whose different subtypes have been proved to regulate different stages of NAFLD, thus becoming promising drug targets for NAFLD. As important sources of drug development, natural products have been proven to treat NAFLD through multiple pathways and multiple targets. In this paper, we outline the regulatory role of PPARs in NAFLD, and summarize some natural products that target PPARs to ameliorate NAFLD, in order to provide reference for drug development of NAFLD.

酒精性肝病(ALD)囊括了一系列由于长期、大量的酒精摄入所致的肝脏疾病,根据疾病发展的进程以及相应的临床特征、病理生理表现,主要分为酒精性脂肪肝、酒精性肝炎及肝纤维化、酒精性肝硬化,严重者可发展至肝细胞癌,大致可与中医学的"酒伤""酒癖""酒疸""酒臌"等病证相对应.目前ALD尚无确切有效的治疗方法,主要以患者禁欲戒酒、营养支持、皮质类固醇、乙酮可可碱、肝移植等为主[1].近年来,许多专家学者对该病的认识逐渐深入,同时也展开了大量的临床及实验研究,成果显著,现试进行梳理.

Background Colorectal carcinoma (CRC) is the third most common cancer and the second most common cause of cancer-related death worldwide. RNA N6-methyladnosine (m6A) and methyltransferase-like 3 (METTL3) play an important role in cancer. However, the roles of m6A and METTL3 in CRC progression are still elusive. Methods Adenoma and CRC samples were applied to detect m6A and METTL3 levels, and tissue microarrays were performed to evaluate their associations with survival of CRC patients. The biological functions of METTL3 were investigated by CCK8, wound healing and transwell assays. M6A epitranscriptomic microarray, RNA stability and luciferase reporter assays were performed to explore the mechanism of METTL3 in CRC. Results m6A and METTL3 levels were significantly upregulated in both adenoma and CRC tissues, and the CRC patients with high m6A or METTL3 level had both shorter overall survival. METTL3 knockdown markedly inhibited the proliferation, migration and invasion of CRC cells. M6A epitranscriptomic microarray revealed that the cell polarity regulator Crumbs3 (CRB3) was the downstream target of METTL3. METTL3 knockdown markedly inhibited the degradation of CRB3 mRNA to increase the CRB3 expression. In addition, CRB3 level was also markedly reduced in both adenoma and CRC tissues, and the CRC patients with high CRB3 level had higher overall survival and disease free survival. CRB3 knockdown significantly promoted the proliferation, migration and invasion of CRC cells. Finally, CRB3 knockdown inhibited Hippo pathway, and increased nuclear localization of YAP. Conclusions The m6A and METTL3 levels were significantly increased in both adenoma and CRC tissues. The CRC patients with high m6A or METTL3 levels had shorter overall survival. Mechanistically, METTL3 regulated the initiation and progression of CRC via regulating m6A-CRB3-Hippo pathway.

目的:探讨苓桂术甘汤对高脂饮食诱导的脂肪肝大鼠脂质代谢相关基因表达的影响,阐释苓桂术甘汤改善高脂饮食诱导的脂肪变性的作用机制.方法:24只大鼠随机分成3组,每组8只.正常组予正常饮食喂养8周;模型组高脂饮食饲养8周;苓桂术甘汤组高脂饮食饲养8周,第5周起同时予苓桂术甘汤(3.6g·kg-1·d-1)灌胃.8周后,取各组大鼠肝脏组织标本,通过RNA测序筛选差异表达基因,并用RT-PCR及Western Blot对差异基因进行验证.结果:模型组Srebp1、Thrsp、Socs2蛋白水平较正常组显著增加(P＜0.01,P＜0.05),而苓桂术甘汤可以显著降低Srebp1、Thrsp、Socs2的蛋白水平(P＜0.01,P＜0.05).结论:苓桂术甘汤可以通过Thrsp-Srebp1通路改善高脂饮食诱导的脂肪变性,而Socs2也是苓桂术甘汤的作用靶点之一.