JOURNAL OF CELLULAR AND MOLECULAR MEDICINE最新文献

High-risk human papillomavirus (HPV) is a central factor in cervical cancer development, largely due to its E6 and E7 oncoproteins that disrupt normal cellular regulation. This study explored the influence of high-risk HPV on the expression of PI3K and the long non-coding RNAs (lncRNAs) MALAT1, H19 and LINC00460 in cervical cells. Using a case–control design, cervical liquid samples from 50 HPV-positive patients and 20 healthy controls were analysed via quantitative real-time PCR, with statistical methods employed to assess correlations between viral oncoproteins and target gene expression. Results demonstrated a significant upregulation of PI3K (24.59-fold change, p < 0.036), MALAT1 (9.75-fold change, p < 0.005), LINC00460 (1.15-fold change, p < 0.013) and H19 (7.1-fold change, p < 0.018) in HPV-infected samples, indicating their potential role in HPV-mediated oncogenesis. Although correlation analysis revealed trends between E6/E7 and certain lncRNAs, these were not statistically significant. Overall, these findings deepen our understanding of the molecular changes linked to high-risk HPV infections and identify PI3K, MALAT1 and H19 as promising biomarkers and therapeutic targets for cervical cancer. Future studies should further investigate these interactions to enhance early detection and improve treatment strategies for HPV-associated malignancies.

Clostridioides difficile (C. difficile ) is a Gram-positive anaerobic bacillus that causes intestinal disorders. Toll-like receptor 4 (TLR4) plays a key role in innate immunity. This study examines the role of TLR4 in the response to C. difficile toxins, which induce cell death and inflammatory responses in enteric glial cells (EGCs). Male C57BL/6 mice were infected with C. difficile, and cecum samples were analysed 3 days post-infection for TLR4 expression. In vitro, EGCs were exposed to C. difficile toxins with or without C34, a TLR4 antagonist, or pre-exposed to TLR4-specific 21-nt small interfering RNAs (siRNA). TLR4 expression was assessed by immunocytochemistry, immunofluorescence, qPCR, and Western blotting. NFκB p65, TNF-α, IL-6, cleaved caspase-3, and phosphatidylserine binding to annexin-V were evaluated. TLR4 expression increased in infected intestinal tissue and toxin-exposed EGCs. TLR4 antagonist or TLR4 knockdown reduced NFκB p65 nuclear translocation and TNF-α expression but did not affect IL-6 upregulation. Additionally, TLR4 antagonist or TLR4 knockdown mitigated toxin-induced cell death, as shown by decreased cleaved caspase-3 and phosphatidylserine binding. These findings suggest that TLR4 contributes to C. difficile pathogenesis and that its inhibition reduces inflammation and prevents cell death in EGCs.

Multiple sclerosis (MS) is a chronic inflammatory disease characterised by myelin and axonal loss. Lack of remyelination is primarily attributed to the failure of oligodendrocyte progenitor cells (OPCs) to differentiate into mature oligodendrocytes. The neuromodulator adenosine can influence OPC differentiation, and by selectively stimulating A_2A and A_2B receptors (A_2AR, A_2BR), it inhibits OPC maturation. In the efforts of developing remyelinating and neuroprotective agents, this study evaluated the ability of a novel dual A_2AR/A_2BR antagonist, P626, in the experimental autoimmune encephalomyelitis (EAE) mouse model and cultured OPCs. EAE mice, 14 days after MOG_35–55 immunisation, received intranasal administration of P626 for 2 weeks, which improved motor symptoms, as evidenced by reduced clinical scores and enhanced performance on the rotarod test, and alleviated thermal and mechanical hypersensitivity without significantly affecting body weight. In spinal cord sections, P626 protected from the reduction of Luxol Fast Blue staining and increased myelin basic protein staining in immunohistochemical analysis. Patch-clamp experiments on cultured OPCs exposed to high extracellular adenosine concentrations demonstrated that P626 prevented the A_2AR- and A_2BR-mediated reduction in sustained I_K and transient I_A currents, both essential for cell differentiation. In conclusion, P626 showed efficacy in reducing neurological symptoms and demyelination in an MS model.

Jiangtang Tiaozhi Formula (JTTZF), a traditional Chinese medicine (TCM) prescription, has been widely used clinically for obesity-related type 2 diabetes (T2D) for many years that can clear heat, release turbidity, open up stagnation and unblock meridians. Several previous clinical studies have demonstrated its effectiveness in decreasing glucose and lipid metabolism disorders, weight loss, and improving chronic inflammation and insulin resistance (IR); however, the exact pathways through which it influences obesity-related T2D require further investigation. This study aims to establish a systematic approach to the pharmacological basis of JTTZF and assess the therapeutic efficacy and its potential mechanisms of JTTZF in ameliorating obesity-related T2D induced by high-fat diet (HFD). Using ultra-high-performance liquid chromatography-mass spectrometry (UHPLC–MS), we identified JTTZF metabolites. Obesity-related diabetic models were established in both mice and zebrafish. The treatment effects were evaluated through haematoxylin and eosin (H&E) and oil Red O (ORO) staining, transmission electron microscopy and assessment of glucose/lipid metabolism indicators. Finally, the specific molecular mechanism underlying JTTZF's efficacy against this condition was comprehensively analysed via in vivo experimental verification. UHPLC–MS/MS identified 371 compounds in JTTZF, with 14 prototype constituents (e.g., demethyleneberberine, epiberberine) absorbed in the liver, linked to anti-diabetic activity. In HFD-induced zebrafish and C57BL/6 mice models, JTTZF significantly ameliorated glucose and lipid metabolic disorders. Histopathological and ultrastructural analyses revealed attenuated hepatic steatosis, reduced lipid droplets and restored mitochondrial integrity. JTTZF also suppressed hepatic inflammation by down-regulating proinflammatory cytokines. Mechanistically, JTTZF inhibited the cyclic GMP–AMP synthase (cGAS)–stimulator of IFN genes (STING) pathway, decreasing phosphorylation of cyclic GMP–AMP synthase–stimulator of type I interferon genes (TBK1) and nuclear factor-κB (NF-κB), while STING inhibitor C-176 and Metformin also displayed similar effects. These findings suggest that JTTZF is a therapeutic agent in inhibiting STING-restored metabolic homeostasis for the management of obesity-related T2D via the cGAS-STING/TBK1/NF-κB pathway.

Multiple sclerosis (MS) is an autoimmune disease characterised by neuroinflammation and neurodegeneration. This study investigates genetic and immunological factors in MS, focusing on microglial regulation. We analysed differentially expressed genes using RNA sequencing from MS lesions (GSE108000) and plaques (GSE227781), validated with cis-eQTL analysis, and integrated Mendelian randomisation (MR), SMR, co-localisation, methylation, and protein–protein interaction (PPI) analyses to assess causal effects on MS risk. We identified five genes—ARHGAP25, HLA-DRB1, MERTK, MS4A6A, and SYK—linked to MS susceptibility. MR revealed that elevated levels of ARHGAP25 (OR = 1.45), HLA-DRB1 (OR = 2.24), MERTK (OR = 1.10), MS4A6A (OR = 1.15), and SYK (OR = 1.13) increased MS risk. SMR confirmed a causal link between HLA-DRB1 and MS, while co-localisation analysis showed shared variants with MS for HLA-DRB1 (100%) and SYK (97.93%). Methylation analysis highlighted 10 sites within HLA-DRB1, and PPI and DrugBank analyses revealed interactions between these genes and multiple proteins or chemicals. This study demonstrates the value of integrating genomic and eQTL data through MR to identify novel MS therapeutic targets, particularly microglial genes, offering potential new avenues for treatment.

Pathological cardiac hypertrophy was an important inducement of heart failure, cardiac arrest, and other diseases. To explore how Vasn knockout induced pathological cardiac hypertrophy, bioinformatics and functional studies illustrated the possible mechanism by clarifying the influence of exosome miRNA on the p-MLC2 signal pathway. B-ultrasound, electrocardiogram, pathological staining, and Q-PCR were used to clarify the changes in typical imaging indexes, pathological indexes, and marker molecules. Exosome sequencing and bioinformatics analysis were carried out to mine key miRNA and signal pathways. Q-PCR, IHC, and WB were used to verify the changes in miRNA and related signal pathways. The changes in heart structure and function were detected by pathological staining, electron microscopy, B-ultrasound, and blood biochemistry in the heart tissues and blood of Vasn knockout mice. Vasn knockout mice showed typical imaging, pathological, and molecular features of PCH. Differential analysis of exosome miRNA showed that let-7g-5p, let-7f-5p, and miR-148a-3p significantly increased in the exosomes of Vasn-knockout mice heart. Bioinformatics analysis showed that let-7g-5p and let-7f-5p targeted the Calm/MLCK/p-MLC2 signal pathway, and miR-148a-3p targeted the Rhoa/ROCK1/p-MLC2 signal pathway. The expression levels of miRNA were significantly up-regulated, but related proteins of signal pathways were significantly reduced in Vasn knockout mice. The structure and function showed obvious damage in Vasn knockout mice. VASN knockout led to pathological cardiac hypertrophy, which may regulate the p-MLC2 signalling pathway through exosomal miRNA.

Wilson disease (WD) is an inherited disorder caused by ATP7B mutations, resulting in toxic copper accumulation primarily in the liver and brain. While copper-induced hepatotoxicity is a hallmark of WD, the mechanisms linking copper overload to liver injury remain unclear. This study aimed to investigate the role of cuproptosis, a copper-dependent form of regulated cell death, in WD pathogenesis and identify key cuproptosis-related genes (CRGs). We utilised ATP7B−/− mice and HepG2 cells to model WD. Liver injury was assessed histologically and biochemically. Transcriptomic analysis identified differentially expressed CRGs, followed by machine learning (LASSO, SVM-RFE) to identify key genes. Functional enrichment and protein validation were performed. Candidate biomarkers were evaluated in WD patient serum and confirmed in the mouse model. ATP7B−/− mice showed marked hepatocellular injury with elevated AST, ALT and LDH. Cuproptosis markers (FDX1, DLST, DLAT, LIAS) were upregulated in both liver tissue and HepG2 cells. Copper exposure decreased cell viability and increased LDH release, exacerbated by Elesclomol and alleviated by Tetrathiomolybdate. Transcriptomics revealed Lox, App, Afp, Alb, Gpc1, Gls were central hub genes. Importantly, SiRNA knockdown of Gpc1, Gls, Lox and App alleviated cuproptosis, supporting their key roles in cuproptosis. Cuproptosis plays a critical role in copper-induced liver injury in WD. Key mediators identified include Gpc1, Gls, Lox and App, which were validated as potential therapeutic targets. These findings provide new insights into the molecular mechanisms underlying WD and may inform the development of targeted treatment strategies.

Diabetic foot ulcers (DFUs) remain a significant clinical challenge due to the lack of effective treatments, severely impacting patients' quality of life. Mesenchymal stem cells (MSCs) have shown potential in promoting DFU healing; however, the underlying mechanisms are not yet fully understood. This study investigates the role of adipose-derived mesenchymal stem cells (ADSCs) in DFU healing, with a particular focus on angiogenesis. Gene expression profiles from the GSE7014 and GSE80178 datasets in the Gene Expression Omnibus (GEO) database were analyzed. Differentially expressed genes (DEGs) were intersected with angiogenesis-related genes from the GeneCards database, identifying 35 angiogenesis-related DEGs (An-DEGs). Key genes were selected using Cytoscape software and machine learning. The pro-angiogenic effects of ADSCs were validated through in vivo and in vitro experiments, assessing their role in DFU healing. The DEGs from DFU patients were enriched in pathways such as angiogenesis and collagen-containing extracellular matrix. ADSCs promoted angiogenesis and wound healing by upregulating FGFR2 and secreting FGF, activating the FGF-PI3K/Akt-HIF-1α-VEGF axis. Additionally, ADSCs mediated secretion of VEGF concerting this effect. FGFR2 plays a pivotal role in ADSCs' mediated DFU healing by driving angiogenesis.

Acute lung injury (ALI) is a life-threatening inflammatory disease of the respiratory system, characterised by high mortality rates and lack of effective clinical interventions. Emerging evidence suggests that traditional Chinese medicine (TCM) formulations may offer therapeutic benefits in managing inflammatory respiratory diseases. Jinbei decoction (JBD), a 12-herb TCM preparation currently used for pulmonary fibrosis, has shown preliminary therapeutic potential in ALI; however, mechanistic studies remain limited. This study systematically evaluated JBD's therapeutic efficacy and elucidated its molecular mechanisms in LPS-induced ALI. Survival analysis demonstrated that JBD significantly improved survival rates in a concentration-dependent manner, while histopathological evaluation revealed a marked reduction in pulmonary tissue damage. These effects were further supported by significant decreases in circulating levels of major pro-inflammatory cytokines, such as TNF-α, IL-6 and IL-1β. Network pharmacology analysis identified 111 molecular targets associated with ALI pathogenesis influenced by JBD components, highlighting the regulatory effect on inflammatory signalling pathways in macrophages as the key intervening mechanism. Specifically, JBD suppressed LPS-induced inflammatory responses by inhibiting ERK phosphorylation and blocking IKKα/β activation, thereby preventing NF-κB-dependent cytokine production in macrophages. Notably, astrapterocarpan was identified as the primary bioactive constituent of JBD through integrated network pharmacology and biochemical analyses. It was found to directly destabilise TRAF6 protein and to exhibit therapeutic efficacy comparable to that of dexamethasone in promoting histological recovery. In vivo experiments further confirmed that JBD significantly reduced TRAF6 expression in murine models, reinforcing the conclusion that its therapeutic effects are predominantly mediated by astrapterocarpan. Collectively, these findings suggest that JBD functions as an agent capable of regulating macrophage polarisation and mitigating cytokine storm through TRAF6-dependent signalling pathways, thereby providing a mechanistic basis for its potential clinical application in inflammatory lung diseases.

Primary biliary cholangitis (PBC) is a chronic disease of a cholestatic nature (more predominant in middle-aged women) resulting from the progressive autoimmune destruction and apoptosis of cholangiocytes in the small intrahepatic bile ducts. This results in the over-accumulation of bile acids (BA), which further damages the bile ducts and hepatocytes [1]. Although PBC is a rare liver condition, if left untreated, it significantly affects the quality of life (jaundice, pruritus, abdominal pain, fatigue) and severely impairs liver function, leading to liver fibrosis, cirrhosis and cancer. UDCA (ursodeoxycholic acid) is the first-line standard treatment of PBC. However, ~40% of patients do not completely respond to the drug. Therefore, obeticholic acid (OCA), a synthetic bile acid that, unlike UDCA, binds to the FXR nuclear receptor, was approved by the FDA in 2016 as a second-line therapy (in monotherapy or associated with UDCA); although OCA has proven anti-fibrotic and anti-inflammatory properties, its efficacy is limited in decompensated cirrhosis and because of side effects (i.e., pruritus) [2].

Among the new therapeutic opportunities, Peroxisome Proliferator-Activated Receptor (PPAR) agonists and SPPARM (selective PPAR modulators) molecules are considered one of the most promising options currently being researched.

The family of PPARs comprises three ligand-activated nuclear receptors—PPARα, PPARβ/δ and PPARγ (encoded by three different genes)—which have emerged as therapeutic targets in metabolic syndrome, type 2 diabetes, dyslipidaemia and inflammation (fibrates and thiazolidinediones have been developed as classical activators of PPARα and PPARγ, respectively). All three PPAR isotypes behave as endogenous fatty acid sensors and transducers of nutritional stimuli into changes in gene expression. Ligand-activated PPARs control the transcription of target genes by binding (as heterodimers with the retinoid-X-receptor RXR, in super-complexes with recruited co-activators) to specific PPAR-responsive elements (PPRE) located in the promoter region of target genes (Figure 1). Therefore, the therapeutic effects of seladelpar (accelerated approval by FDA/2024 and EMA/2025) and elafibranor (accelerated approval by FDA/2024) rely on the regulation (up- or down-regulation) of gene sets controlling metabolic, anti-inflammatory and anti-fibrotic mechanisms (Figure 1).

Until recently, PPARβ/δ was not a target of a marketed drug, but several synthetic ligands (e.g., GW501516, GW7042, L165041) have been developed over time and widely used in preclinical research (and some of them also in clinical trials) to understand the mechanisms and therapeutic impact of PPARδ activation in various tissues. PPARδ is ubiquitously expressed, with relatively high levels in metabolically active tissues—muscle, adipose tissue and liver (hepatocytes, Kupffer cells and cholangiocytes), where it exerts