Journal of Molecular Histology最新文献

Changes in integument pigmentation in fish can occur due to trauma, parasite attachment, or bacterial, viral, and fungal skin infections. Experimental wound-healing models have proven to be valuable tools for studying these processes. This study aims to characterize the chromatophores of Gymnotus carapo and their role in the healing process, using an experimental wound-healing model. The analysis focused primarily on melanophores due to their predominance in the species’ integument; other pigment cell types were not detected in the examined tissues. A total of 25 adult banded knifefish specimens were collected using fishing nets from natural lentic environments in San Cosme, Corrientes, Argentina. The specimens were transported to the Faculty of Veterinary Sciences (FCV) and housed in containers. Lesions were then induced in the mid-dorsal integument of 20 specimens, while 5 were kept as uninjured controls. At 12 hours, 24 hours, 6 days, and 9 days post-injury, five specimens from each group were randomly selected and euthanized in accordance with bioethical guidelines. Macroscopic observations of the integument were conducted using scanning electron microscopy, and histological slides were prepared and stained with Hematoxylin and Eosin and PAS histochemical reactions.Morphological and distributional changes in the chromatophores of G. carapo were observed throughout the healing process, with macroscopic and microscopic variations analyzed at different time points. These findings highlight, for the first time, the dynamic participation of chromatophores during tissue regeneration in this Neotropical species, emphasizing their potential role beyond pigmentation. This research provides new insights into the cellular mechanisms of fish skin repair and establishes a foundational model for studying chromatophore-mediated healing, with implications for improving health and welfare management in aquaculture.

Non-small cell lung cancer (NSCLC) is a leading cause of cancer-related deaths worldwide. Adipocyte Plasma Membrane Associated Protein (APMAP) is a glycosylated type II transmembrane protein, and APMAP induces malignant metastasis of colorectal cancer. However, the role and mechanism of APMAP in NSCLC remain unknown. We aimed to study the APMAP regulation on NSCLC. APMAP mRNA levels in NSCLC tissues were tested by quantitative reverse transcription-PCR (qRT-PCR). APMAP protein levels in NSCLC tissues and cells were determined via Western blot. Also, the association between APMAP expressions and NSCLC clinicopathology was assessed using Chi-square test. After silencing APMAP (sh-APMAP) in NSCLC cells, APMAP’s roles in NSCLC were revealed by Western blot, Cell Counting Kit-8 (CCK-8) assay, Transwell, Fe²⁺ level analysis, and immunofluorescence assays. Meanwhile, APMAP mechanism in NSCLC was verified through Human TFDB and PROMO databases, qRT-PCR, Western blot, dual-luciferase reporter assay, CCK-8 experiment, Transwell, analysis of Fe²⁺, reactive oxygen species levels, and Clinical Bioinformatics Home. Also, APMAP function in vivo was examined using a tumor xenograft model, immunohistochemistry assay, and Western blot. APMAP expressions were up-regulated in NSCLC tissues and cells. Moreover, patients with high APMAP expression exhibited a poorer overall survival rate. APMAP expression was closely related to TNM stage, distant metastasis, and tumor differentiation. Functionally, APMAP knockdown repressed NSCLC cell proliferation and invasion, and induced cell ferroptosis. Mechanistically, transcription factor YY1 induced APMAP expression. Meanwhile, APMAP expressions were decreased after interfering with YY1, while APMAP expressions were increased after overexpressing YY1. Also, co-transfection experiments demonstrated that YY1 overexpression enhanced the fluorescence intensity of APMAP-WT promoter. Importantly, YY1 overexpression promoted NSCLC cell proliferation and invasion, and repressed cell ferroptosis, yet these effects were reversed after APMAP knockdown. Moreover, knocking down APMAP reduced NSCLC proliferation in vivo, mainly through reduced tumor volume, reduced KI-67 and GPX4 expressions, and decreased p-pI3K, p-AKT, and p-mTOR protein levels. APMAP regulated by YY1 transcription enhanced NSCLC proliferation, invasion, and repressed cell ferroptosis via activating PI3K/AKT/mTOR.

Graphical abstract

The highly expressed YY1 induces the high APMAP expression, promotes NSCLC cell proliferation and invasion, and represses cell ferroptosis via activating PI3K/AKT

Intervertebral disc degeneration (IDD) is the main cause of low back pain and is related to the aging of nucleus pulposus (NP) cells (NPCs). However, the underlying mechanism for senescence of NPCs has not been well understood. NPCs (control-NPCs and IDD-NPCs) were separated from the NP tissues of patients, diagnosed with graded I and IV IDD. The proliferation and senescence levels were detected by MTT assay and senescence-associated β-galactosidase staining, respectively. The expression of cyclin B1 and caveolin-1 was evaluated by Western blot analysis. RNA-sequencing technology was used to screen differentially expressed genes (DEGs) in IDD-NPCs. Finally, through the interference and overexpression of MYB proto-oncogene-like 2 (MYBL2), the effect of MYBL2 on cell senescence was explored. Compared with control-NPCs, the proliferation ability of IDD-NPCs was significantly reduced and the senescence level was obviously increased. 1,061 DEGs were screened in IDD-NPCs. By analyzing DEGs related to aging and aging-related pathways and terms, cell cycle arrest was identified. Knockdown of MYBL2 promoted the senescence of control-NPCs, and overexpression of MYBL2 inhibited the senescence of IDD-NPCs. The study found that upregulation of MYBL2 ameliorated the senescence of IDD-NPCs, providing a potential way to inhibit the senescence of NPCs.

Slow transit constipation (STC) is a prevalent functional gastrointestinal disorder characterized by a reduced frequency of bowel movements, the presence of dry and hard stools, and abdominal pain. However, the underlying mechanisms contributing to its pathogenesis have not yet been fully clarified. This study aims to investigate the effects of METTL3 on loperamide (LOP)-induced STC mice and glutamic acid-induced interstitial cells of Cajal (ICCs). METTL3-knock down adeno-associated virus (AAV) was used to treat LOP-induced mice, and the effect of METTL3 down-regulation was assessed by the stool parameters, histological analysis, transmission electron microscopy (TEM), TdT-mediated dUTP nick end labeling (TUNEL) staining, immunohistochemistry, Immunofluorescence staining, and Western blotting. METTL3 small interfering RNA (siRNA) was transfected into ICCs before glutamic acid, PI3K inhibitor (LY294002), and AKT inhibitor (GSK690693) treatment alone or in combination. EdU assays, flow cytometry, TEM, and Western blot were used to investigate the relationship between METTL3 and PI3K/AKT pathway. METTL3 deletion alleviated constipation symptoms and promoted intestinal motility in STC mice. METTL3 knockdown suppressed apoptosis and autophagy, accompanied by increased proliferation of glutamic acid-induced ICCs. More importantly, the effect of METTL3 knockdown on proliferation and autophagy was significantly reversed in glutamic acid-induced ICCs treated with LY294002 or GSK690693. Mechanistically, METTL3 deletion exerts its STC-repressive influence through the activation of the PI3K/AKT pathway. Collectively, the findings indicate that METTL3 modulates PI3K/AKT-mediated autophagy following LOP and highlight the potential of METTL3 as a therapeutic target in STC treatment.

In an era where precision medicine demands rapid innovation, molecular docking has emerged as a cornerstone technology that transforms the study of histological biomarkers and disease biology. This review provides a comprehensive overview of molecular docking techniques applied to the exploration of histological biomarkers across diverse disease models. Contemporary approaches such as AutoDock, FlexX, SwissDock, PyRx, Surflex, and ICM, as well as molecular dynamics simulations are discussed in relation to their role in identifying and validating ligand interactions with key protein targets, including SREBP, PPAR-α, FXR, MAO-B, α-synuclein, and HAT. The utility of docking is examined in the context of liver fibrosis, neurodegenerative diseases, cancer, and metabolic disorders, highlighting how in silico screening accelerates the discovery of potential therapeutics such as silymarin, ursolic acid, quercetin, berberine, and diallyl trisulfide. Studies integrating docking with in vitro and in vivo validation demonstrate improved targeting of disease-relevant proteins and the prediction of binding affinities, supporting personalized medicine and drug repurposing. Virtual screening reduces experimental workload by efficiently identifying promising candidates. It also elucidates multi-target interactions and enhances understanding of underlying mechanisms, as demonstrated by data from recent literature. Overall, molecular docking emerges as a crucial tool for mapping disease mechanisms, prioritizing therapeutic candidates, and guiding biomarker-driven drug development in modern biomedical research.

Lipid disorder is an independent risk factor of diabetic kidney disease (DKD). Excess accumulation of lipid in podocytes can cause cell dysfunction and cell death. Chaperone-mediated autophagy (CMA) serves as a critical role in regulating lipid metabolism. However, the exact role of CMA in the podocytes of DKD with dyslipidemia is still uncertain. Herein, we aimed to explore the role of CMA in hyperlipidemia-induced lipid accumulation and apoptosis in podocytes. In the present study, we showed that palmitic acid (PA) treatment induced the activation of CMA, increased lipid accumulation and apoptosis in podocytes. We further found that blocking CMA with inhibitor VER155008 or LAMP-2 A siRNA significantly upregulated PA-induced increased expression of PLIN2, exacerbated PA-induced lipid accumulation and apoptosis, whereas promoting CMA with Torin1 downregulated the expression of PLIN2, ameliorated lipid accumulation and apoptosis in PA-induced podocytes. Moreover, we also observed the activation of CMA and increased lipid accumulation in the kidney tissue of DKD mice. Taken together, these results suggest that CMA plays a protective role in PA-induced podocytes apoptosis and that the potential protective mechanism of CMA is involved in reducing cellular lipid accumulation through mediating the degradation of PLIN2.

Yin Yang 1 (YY1) is a key transcription factor that drives prostate cancer (PCa) progression through bidirectional transcriptional regulation. Clinically, high expression of YY1 and abnormal cytoplasmic aggregation in PCa tissues are significantly associated with aggressive clinical-pathological features. These patterns hold important diagnostic and prognostic value at the tissue level, serving as promising histopathological biomarkers for assessing tumor invasiveness and therapeutic resistance. Mechanistically, YY1 drives PCa progression through multifaceted networks: it transcriptionally activates oncogenes such as Krüppel-like factor 4 (KLF4) and Prostate Specific Antigen (PSA); epigenetically induces EMT via Twist1/hnRNPM and stimulates YAP/TAZ signaling through the DNAH8AS1/miR-186-5p axis; promotes metabolic reprogramming by synergizing with BRD2/4 to activate PFKP and facilitating hypoxia adaptation via phase separation with HIF-1α; and contributes to immunosuppression by polarizing M2 macrophages and upregulating PD-L1 through IL-4/STAT6, thereby inhibiting CD8⁺ T cells. YY1 also sustains prostate cancer stem cells (PCSCs) self-renewal and chemoresistance via SOX2/BMI1. Targeting strategies for YY1 include small-molecule inhibitors (e.g.,Tenapanor), epigenetic drugs (e.g., Entecavir), gene editing, and immune combination therapies. By disrupting interactions between YY1 and HIF-1α/HDAC1/BRD4 or reversing immune suppression and metabolic adaptation, these approaches significantly inhibit tumor progression. This review systematically outlines the multidimensional regulatory mechanisms and therapeutic targeting of YY1 in PCa, offering a foundation for precision transcription factor intervention.

Bone marrow mesenchymal stem cells (BMSCs) play a significant role in bone defect repair. Currently, the limited survival and function of transplanted BMSCs pose major obstacles to tissue repair mediated by these cells. Therefore, the exploration of BMSCs transplantation activity has become an urgent clinical problem. In this experiment, we constructed lentiviral vectors with overexpression and silencing of miR-19b-3p and utilized a rat fracture model. The effects of BMSCs proliferation were examined using CCK-8, the Annexin V-FITC apoptosis kit and TUNEL assay, the effects of BMSCs osteogenic differentiation were detected using ALP and Alizarin red staining. Furthermore, Western blotting and qRT-PCR were employed to examine the expression of key proteins in the PTEN/Akt/mTOR signaling pathway and autophagy-related proteins. Moreover, the impact of miR-19b-3p on bone microstructure was assessed using Micro CT, HE, Safranin O/Fast Green, as well as Masson staining, and Immunohistochemistry analysis examined PTEN, p62 and Runx2 expression levels on fracture site. By constructing lentiviral vectors with overexpression and silencing of miR-19b-3p, we found that silencing miR-19b-3p significantly promoted the proliferation as well as the osteogenic differentiation of BMSCs under hypoxia. Western blotting and qRT-PCR results showed that silencing miR-19b-3p significantly increased PTEN, LC3, and Beclin1 expression while decreasing p-Akt, p-mTOR, and p62 expression. Furthermore, by constructing the fracture rats and administering hypoxia-treated BMSCs transfected with miR-19b-3p via tail vein injection, we found that silencing miR-19b-3p obviously increased the formation of trabecular and promoted the healing of the fractures. Immunohistochemistry analysis showed that silencing miR-19b-3p upregulated PTEN and Runx2 expression levels and suppressed p62 expression with statistical significance. In summary, these findings suggest that miR-19b-3p may regulate BMSCs’ autophagy and osteogenic differentiation under hypoxia via the PTEN/Akt/mTOR signaling pathway, thereby promoting fracture healing and providing a potential target for the treatment of fractures.

Skeletal muscle exhibits remarkable plasticity, allowing it to adapt to varying metabolic and regenerative demands. However, conditions such as myopathies compromise its integrity. To study muscle degeneration and regeneration, various models have been developed, including chemical injury induced by BaCl₂. This agent causes myofiber damage similar to that produced by certain biological toxins, yet it spares satellite cells, thereby enabling regeneration. Key regulators such as MyoD and active β-catenin are essential for activating myogenic pathways during repair, although their behavior may vary across muscle types. Currently, there is limited information on effective treatments for many myopathies. Epicatechin (Epi), a natural flavonoid, has shown potential to reduce fibrosis and promote myogenic protein expression. In this study, we evaluated the effect of Epi on gastrocnemius muscle repair following BaCl₂-induced injury in CD1 mice. A total of 60 mice were divided into four groups (n = 3 for morphology and gene expression analysis, and n = 6 for western blot analysis). The results showed that Epi reduced the injury area compared to the untreated groups. Active β-catenin and MyoD levels increased within 1 h post-injury, while Myogenin expression continued to rise up to 24 h. Interestingly, the non-injured plus Epi group also showed increased levels of MyoD and Myogenin, as well as increased cross-sectional area (CSA). Additionally, Epi appeared to influence the expression of myosin heavy-chain isoforms and to induce differential hypertrophy in type II muscle fibers, findings warranting further investigation. Moreover, (–)-Epicatechin accelerated muscle repair and reduced muscle injury, suggesting its potential as a supportive agent in muscle repair therapies.

Triple-negative breast cancer (TNBC) is highly malignant with rising incidence and mortality. Solute carrier family 7 member 5 (SLC7A5) is an amino acid transporter, and its mechanism in TNBC is still unclear. The public databases (TIMER2.0, TCGA, GEPIA, Kaplan-Meier Plotter, linkedomics, RBPmap, and RBPDB) were used to analyze the expression and correlation of genes and prognosis correlation. Gene and protein expression was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot. 3-(4, 5)-Dimethylthiahiazo (-z-y1)-3, 5-di-phenytetrazoliumromide (MTT), terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL) staining, flow cytometry, transwell, sphere/tube formation, and metabolic kits were used to assess cell viability, apoptosis, invasion, stemness, angiogenesis, and glutamine (Gln) metabolism. The binding of forkhead box M1 (FOXM1) to the SLC7A5 promoter was determined by dual-luciferase reporter assay/chromatin immunoprecipitation (ChIP). RNA binding protein immunoprecipitation (RIP), RNA pull-down, and actinomycin D (Act D) experiments evaluated fused in sarcoma (FUS)-SLC7A5 interaction. In vivo, the mouse xenografts were established to validate the effects of FOXM1/SLC7A5 axis. Hematoxylin-eosin (HE) and immunohistochemistry (IHC) assays were used for histological morphology analysis and the protein expression of Ki67 and SLC7A5. SLC7A5 was up-regulated in TNBC and correlated with poor prognosis. Its knockdown repressed TNBC cell viability, invasion, stemness, angiogenesis, and Gln metabolism (as evidenced by reduced Gln, α-ketoglutarate (α-KG), and adenosine 5’-triphosphate (ATP) levles). FOXM1 transcriptionally activated SLC7A5; SLC7A5 overexpression reversed the suppressive effects of FOXM1 knockdown on TNBC malignancy and metabolism. Additionally, FUS bound to and stabilized SLC7A5 mRNA. In vivo, SLC7A5 counteracted the FOXM1 knockdown-mediated inhibition of tumor growth and the reduction in SLC7A5 and Ki67 protein expression. In conclusion, SLC7A5 promotes TNBC malignancy and metabolism. Its expression is transcriptionally driven by FOXM1 and post-transcriptionally stabilized by FUS. Targeting the FOXM1/SLC7A5 axis presents a novel therapeutic strategy for TNBC.