Sheng Cheng

Background: Liver fibrosis involves extensive metabolic remodeling, but the coordinated changes in proteins, metabolites and cell-type-specific gene expression signatures remain incompletely defined. This study aimed to comprehensively characterize the molecular and cellular metabolic alterations in liver fibrosis and identify potential metabolic targets for antifibrotic therapy. Methods: A CCl ₄ -induced murine hepatic fibrosis model was established and validated by histology and serum markers. Integrated proteomics, metabolomics, and single-cell transcriptomics were performed to profile molecular and metabolic changes. Pathway enrichment (KEGG, GO, MBROLE) and network analyses, including Ingenuity Pathway Analysis (IPA), were conducted. Functional validation in hepatic stellate cells (HSCs) included western blot, qPCR, immunofluorescence, and Seahorse assays. Public datasets from metabolic-associated steatohepatitis (MASH/MASLD) patients were analyzed for translational relevance. Results: Integrated multi-omics analysis identified coordinated disorders of arachidonic acid, retinol, linoleic acid, arginine-proline and butanoate metabolism in fibrotic livers. IPA network analysis highlighted PPARα and ERK1/2 as hub regulators, and pharmacological intervention confirmed their direct participation in HSCs activation. Liver fibrosis was characterized by widespread suppression of hepatocyte cytochrome P450 enzymes, disrupted arachidonic acid and retinoid metabolism, and accumulation of glycerophospholipids including phosphatidylethanolamine (PE). Although total PE levels rose in fibrotic liver tissue, exogenous POPE significantly inhibited HSCs activation. Mechanistically, POPE suppressed the MAPK pathway, upregulated PPARα, and restored excessive mitochondrial respiration and glycolysis in activated HSCs. Cell-type-resolved profiling showed that extracellular matrix genes were mainly expressed in HSCs, adhesion and cytoskeletal genes in macrophages, and downregulated cytochrome P450 genes in hepatocytes. These metabolic molecular alterations were consistent with transcriptomic trends in human liver samples with advanced fibrosis. Conclusions: Liver fibrosis is characterized by coordinated, cell-type-dependent remodeling of lipid and energy metabolism. Hepatocyte-associated cytochrome P450 suppression, altered HSC bioenergetics and dysregulated glycerophospholipid metabolism collectively contribute to the fibrotic metabolic landscape. POPE was associated with reduced HSC activation and normalization of MAPK, PPARα and bioenergetic responses, identifying defined phosphatidylethanolamine species as potential modulators of HSC state. Further mechanistic and in vivo studies are required to determine the causal and therapeutic relevance of these findings.

Purpose: This study aims to explore the potential mechanisms of esculin in the treatment of renal cell carcinoma (RCC). Methods: We employed network pharmacology to predict the potential mechanisms and targets of esculin in RCC. Molecular docking techniques were then employed to validate the predicted targets. Additionally, a series of in vitro experiments were conducted to verify the anticancer effects of esculin on RCC cells, including the CCK-8 assay, EdU assay, wound healing assay, apoptosis assay, and Western blot. Results: Network pharmacology and molecular docking results identified GAPDH, TNF, GSK3B, CCND1, MCL1, IL2, and CDK2 as core targets. GO and KEGG analyses suggested that esculin may influence apoptotic processes and target the PI3K/Akt pathway in RCC. Furthermore, the CCK-8 assay demonstrated that esculin inhibited RCC cell viability. Microscopic observations revealed that following esculin treatment, there was an increase in cell crumpling, a reduction in cell density, and an accumulation of floating dead cells. Additionally, with increasing esculin concentrations, the proportion of EdU-positive cells decreased, the wound closure ratio decreased, the proportion of PI-positive cells increased, the expression levels of BAX and cleaved-caspase-3 proteins increased, and the expression level of Bcl2 protein decreased. These findings suggested that esculin inhibits the proliferation and migration of RCC cells while promoting apoptosis. Moreover, esculin was found to target GAPDH and inhibit the PI3K/Akt pathway. Conclusions: This study is the first to elucidate the therapeutic effects of esculin on RCC cells. The results provide evidence supporting the clinical application of esculin and introduce a promising new candidate for RCC treatment.

Hepatocellular carcinoma (HCC) still presents poor prognosis with low overall survival rates and limited therapeutic options available. Recently, attention has been drawn to peptidomic analysis, an emerging field of proteomics for the exploration of new potential peptide drugs for the treatment of various diseases. However, research on the potential function of HCC peptides is lacking. Here, we analyzed the peptide spectrum in HCC tissues using peptidomic techniques and explored the potentially beneficial peptides involved in HCC. Changes in peptide profiles in HCC were examined using liquid chromatography-mass spectrometry (LC–MS/MS). Analyze the physicochemical properties and function of differently expressed peptides using bioinformatics. The effect of candidate functional peptides on HCC cell growth and migration was evaluated using the CCK-8, colony formation, and transwell assays. Transcriptome sequencing analysis and western blot were employed to delve into the mode of action of potential peptide on HCC. Peptidomic analysis of HCC tissue yielded a total of 8683 peptides, of which 452 exhibited up-regulation and 362 showed down-regulation. The peptides that were differentially expressed, according to bioinformatic analysis, were closely linked to carbon metabolism and the mitochondrial inner membrane. The peptide functional validation identified a novel peptide, PDLC (peptide derived from liver cancer), which was found to dramatically boost HCC cell proliferation through the Ras/Raf/MEK/ERK signaling cascade. Our research defined the peptide’s properties and pattern of expression in HCC and identified a novel peptide, PDLC, with a function in encouraging HCC progression, offering an entirely new potential therapeutic target the disease.

Background. Hepatocellular carcinoma (HCC) is one of the most prevalent malignancies and is associated with high mortality. Transmembrane protein 147 (TMEM147) is a seven‐transmembrane protein that may mediate immune regulation. However, the relevance of TMEM147 to immune regulation in HCC and the prognosis of HCC patients are unclear. Methods. We analyzed TMEM147 expression in HCC by using the Wilcoxon rank‐sum test. Real time quantitative PCR (RT‐qPCR) and Western blot analysis of tumor tissues and cell lines were used to verify TMEM147 expression in HCC. The influence of TMEM147 on HCC prognosis was assessed using Kaplan–Meier analysis, Cox regression analysis, and a prognostic nomogram. The functions of the TMEM147‐related differentially expressed genes (DEGs) were identified by Gene Ontology (GO)/Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses and gene set enrichment analysis (GSEA). In addition, we examined the associations between TMEM147 expression and immune infiltration using single‐sample gene set enrichment analysis (ssGSEA) and immunofluorescence staining of HCC tissues. Results. Our results showed that the expression of TMEM147 was significantly higher in human HCC tissues than in adjacent normal liver tissues, with similar findings in human HCC cell lines. High TMEM147 expression was correlated with T stage, pathological stage, histological grade, race, alpha‐fetoprotein level, and vascular invasion in HCC. Moreover, we revealed that high TMEM147 expression was associated with shorter survival times and that TMEM147 could be a risk factor for overall survival, along with T stage, M stage, pathological stage, and tumor status. Mechanistic studies revealed that high TMEM147 expression was linked to the B lymphocyte, antigen response, IL6 signaling pathway, cell cycle, Kirsten rat sarcoma viral oncogene homolog (KRAS) signaling pathway, and myelocytomatosis oncogene (MYC) targets. Correspondingly, TMEM147 expression was positively associated with the infiltration of immune cells, including Th2 cells, follicular helper T cells, macrophages, and NK CD56 bright cells in HCC. Conclusions. TMEM147 might be a biomarker for poor prognosis and is related to immune cell infiltration in HCC.