Journal of Molecular Histology最新文献

Prolonged healing of skin wounds remains a major clinical challenge. This study employed network pharmacology combined with in vivo experimental validation to investigate the mechanisms of Panax notoginseng (PN) in treating skin wounds. Network pharmacology identified 156 targets and 115 signaling pathways, with top core targets (TNF, IL-6, IL-10) enriched in NF-κB, MAPK, and JAK-STAT pathways. Molecular docking showed favorable binding affinities between PN components (Ginsenoside Rb₁, Rd, Re, Rg₁, Notoginsenoside R₁) and key target proteins. In vivo, 48 SD rats were randomly divided into a PN group and a control group with a full-thickness skin excision model. The PN group showed significantly smaller wound areas at days 4 and 7 (P < 0.05), with reduced inflammatory cell infiltration and enhanced fibroblast proliferation at day 7. qRT-PCR and ELISA demonstrated that TNF-α was significantly lower in the PN group at days 4 and 7 (P < 0.01), IL-6 was lower at all time points (P < 0.01), and IL-10 was lower at days 1 and 7 (P < 0.01). These findings suggest that PN modulates the inflammatory cytokine network (TNF-α, IL-6, IL-10) in a time-dependent manner, consistent with the regulation of the “inflammation initiation → repair transition” temporal sequence, representing a promising natural agent for regulating the inflammatory phase of skin wound healing.

Prostate cancer ranks second among the most common malignant neoplasms in men worldwide. Fruit extracts with potential anti-prostate cancer properties have gained increasing attention, with the most notable belonging to the Annonaceae family, including Araticum (Annona crassiflora Mart). In this study, we aimed to evaluate the chemopreventive and anti-inflammatory potential of Araticum seed extract (Annona crassiflora Mart) on the ventral prostate lobe of Transgenic Adenocarcinoma of the Mouse Prostate (TRAMP) model. Sixty male TRAMP mice, aged 8 and 12 weeks, were divided into four groups: two groups received the extract at a dose of 100 mg/kg body weight, and two control groups received equivalent volumes of water and DMSO (Dimethyl Sulfoxide). After 30 days, animals were euthanized and ventral prostate samples were collected for morphological evaluation by light microscopy, immunohistochemistry, and Western blotting. Treatment with Araticum seed extract delayed the progression of premalignant and malignant lesions (HGPIN and adenocarcinoma) and preserved healthy tissue. The extract showed antiproliferative activity, reducing PCNA, IGFR-1, and AR protein levels. Inflammatory signaling was suppressed via Toll-like receptor pathways, with decreased NFκB, TNF-α, and other inflammatory markers. Moreover, the extract reduced BCL-2 expression and promoted caspase activation, suggesting pro-apoptotic protection. In conclusion, treatment with Araticum seed extract demonstrated promising chemopreventive activity, effectively reducing the progression of epithelial lesions in the ventral prostate lobe of TRAMP mice and may be suggested as a potential chemopreventive agent in combined prostate cancer therapies.

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Chronic kidney disease (CKD) is associated with sensorineural hearing loss, but the mechanisms linking renal injury to cochlear dysfunction remain incompletely defined. Because lipocalin-2 (LCN2/NGAL) is markedly increased in CKD and has been implicated in endothelial dysfunction and barrier injury, we investigated whether LCN2 contributes to cochlear damage in a mouse model of CKD. Male C57BL/6J mice underwent 5/6 nephrectomy, and CKD mice received either an anti-LCN2 monoclonal antibody or an isotype control three times weekly for seven weeks beginning one week after completion of surgery. Auditory function, cochlear morphology, blood–labyrinth barrier integrity, cochlear perfusion, and LCN2-associated signaling were assessed using auditory brainstem responses, distortion product otoacoustic emissions, whole-mount immunofluorescence, FITC–dextran extravasation, Western blotting, quantitative PCR, and laser Doppler flowmetry. CKD mice developed elevated auditory brainstem response thresholds, reduced distortion product otoacoustic emission amplitude at a single primary tone pair with f2 = 8.0 kHz, outer hair cell loss, and a reduction in the number of CtBP2-positive presynaptic ribbon puncta per inner hair cell. These changes were accompanied by increased serum and cochlear LCN2, upregulation of 24p3R/SLC22A17 and NLRP3, increased blood–labyrinth barrier permeability, reduced claudin-5 and ZO-1 expression, impaired cochlear blood flow, stria vascularis atrophy, and decreased expression of strial ion-transport genes. Anti-LCN2 antibody treatment improved auditory thresholds and otoacoustic emission responses, attenuated outer hair cell and synaptic injury, reduced vascular leakage, partially restored tight junction and ion-transport gene expression, and improved cochlear perfusion, without measurably lowering serum blood urea nitrogen or creatinine. These findings support a possible contribution of LCN2 to CKD-associated cochlear vascular dysfunction and hearing loss. Targeting LCN2 may represent a therapeutic strategy for preserving hearing in CKD, although validation in additional CKD models, both sexes, and human tissues is required.

Acampe papillosa (Lindl.) Lindl. has long been used to cure a variety of illnesses, including rheumatism and traumatic tissue injuries. Nevertheless, there is a dearth of comprehensive information on its phytochemical composition and pharmacological actions. Therefore, here, we present the first thorough pharmacological and histopathological analysis of Acampe papillosa seed methanolic extract (ME-APS), together with in silico molecular docking studies, hepatoprotective, wound-healing, and anti-inflammatory properties. Before pharmacological evaluations, safety was assessed through oral and dermal toxicity studies. The anti-inflammatory activity of ME-APS was tested using carrageenan and formalin-induced paw oedema models, and the wound healing potential was observed using incision and excision wound models. Hepatoprotective activity was studied in the CCl₄-induced hepatotoxicity model in rats. In oral (up to 5000 mg/kg) and dermal (10% ointment formulation) toxicity studies, ME-APS was found to be safe with no visible signs of toxicity. In the study on anti-inflammatory properties, ME-APS significantly reduced paw oedema in carrageenan and formalin models of inflammation. Wound healing assays showed that the extract significantly improved wound contraction, tensile strength, epithelialization, collagen deposition, and fibroblast proliferation in a dose-dependent way. The hepatoprotective study showed that ME-APS treatment significantly restored the altered biochemical parameters, such as ALT, AST, TC, TG, and LDL levels, and ameliorated CCl₄-induced histopathological liver damage. Further, the molecular docking study showed that the major phytochemicals in the extract, such as squalene, cholesta-4,6-dien-3-ol (3β), stigmasterol, and γ-sitosterol, showed significant binding affinities for COX-2 and TGF-β–related targets involved in inflammation, wound healing, and hepatoprotection. Additionally, a high probability of biological efficacy was predicted using PASS (Prediction of Activity Spectra for Substances) analysis. Overall, the findings suggest that ME-APS has possible anti-inflammatory, wound healing, and hepatoprotective activities that support its traditional medicine use and imply that it is a potential source of therapeutic phytochemicals.

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Although ferroptosis contributes to tumor progression, the key regulators of this process in nasopharyngeal carcinoma (NPC) remain largely unknown. TRIM26, an E3 ubiquitin ligase, has not been systematically studied in NPC or ferroptosis. Ferroptosis-related NPC genes were screened using GeneCards, FerrDb, and UbiBrowser2.0 databases. TRIM26 expression in NPC cells was determined via RT-qPCR and Western blot. TRIM26 overexpression models were established in C666-1 and NPC/HK1 cells, and a ZEB1 rescue model was further constructed in C666-1 cells. Cell viability, migration, and invasion were tested using CCK-8, wound healing, and Transwell assays. Ferroptosis was evaluated using C11-BODIPY581/591 probe, malondialdehyde (MDA), glutathione (GSH), and Fe²⁺ measurements. Co-immunoprecipitation, ubiquitination, and cycloheximide chase assays were performed to investigate TRIM26 regulation of ZEB1. Dual-luciferase reporter assays and ChIP-qPCR were conducted to verify the transcriptional regulation of SLC7A11 by ZEB1. In vivo validation was conducted using a subcutaneous xenograft model. TRIM26 was significantly downregulated in NPC cells, most markedly in C666-1 cells. TRIM26 overexpression remarkably inhibited NPC cell viability, migration, and invasion. It markedly promoted ferroptosis, with increased lipid peroxidation, elevated MDA and Fe²⁺ levels, reduced GSH content, and downregulated GPX4 and SLC7A11. Mechanistic studies revealed that TRIM26 interacted with ZEB1, promoted its ubiquitination, and accelerated its protein degradation. Furthermore, ZEB1 directly regulated SLC7A11 transcription through binding to its promoter region. ZEB1 overexpression significantly reversed the ferroptotic phenotype induced by TRIM26. In vivo, TRIM26 overexpression inhibited xenograft tumor growth and activated ferroptosis, whereas co‑overexpression of ZEB1 partially attenuated these tumor-suppressive effects. TRIM26 activates ferroptosis by promoting the ubiquitination and degradation of ZEB1 protein, thereby inhibiting NPC growth and progression. The TRIM26-ZEB1 axis may represent a potential molecular target for NPC.

Mastitis is a major inflammatory disorder of the mammary gland that causes substantial economic losses in the dairy industry, with Staphylococcus aureus (S. aureus) being one of the predominant causative pathogens. The current treatment options are limited by antimicrobial resistance and drug residues. Therefore, alternative therapeutic strategies are needed. The present study aimed to evaluate the anti-inflammatory and antioxidant effects of an ethnoveterinary oil (EO) formulation in an experimental model of S. aureus–induced mastitis. The EO formulation was prepared according to a standardized traditional ethnoveterinary composition. Mastitis was induced in healthy lactating primiparous Wistar rats by intramammary infusion of S. aureus (2 × 10⁸ CFU/mL), and the twenty-four female Wistar rats were divided into four groups of 6: normal control (NC), mastitis control (MT), standard drug–treated (MT + DEX), and MT + EO-treated groups. The chemical composition of EO was characterized by GC–MS analysis, which identified 21 bioactive compounds, predominantly erucic acid, along with phenolic compounds, fatty acid derivatives, terpenoids, and steroidal constituents known for their anti-inflammatory potential. Biochemical analyses revealed that EO significantly restored endogenous antioxidant enzyme activities and reduced lipid peroxidation in mammary tissues. Furthermore, EO significantly decreased myeloperoxidase (MPO) levels and N-acetyl-β-D-glucosaminidase activity, indicating reduced neutrophil infiltration and protection against epithelial cell damage. The levels of Pro-inflammatory cytokines were markedly elevated in mastitis control group whereas EO treatment significantly suppressed their production. In addition, EO downregulated the gene expression of Toll like receptor-2 (TLR-2) and Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 2 (NOX-2), suggesting modulation of key inflammatory and oxidative stress–related pathways. Histopathological and special staining analyses further confirmed the preservation of mammary gland architecture and attenuation of inflammatory infiltration following EO treatment. Collectively, these findings demonstrate that EO exerts potent anti-mastitic effects through the combined modulation of oxidative stress and inflammatory responses and may serve as a promising natural therapeutic alternative for the management of S. aureus associated mastitis in Wistar rat model.

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Radiotherapy is an effective treatment for head and neck cancer; however, it also induces alterations in healthy oral tissues. Although late radiation-induced damage has been extensively investigated, the early progression of morphological and extracellular matrix changes in the periodontal ligament (PDL) remains poorly understood. This study investigated early radiation-induced changes in collagen organisation, the type I-to-type III collagen ratio, and matrix metalloproteinase-2 (MMP-2) expression in the lingual PDL of rat incisors. Adult Wistar rats were allocated to four irradiated groups (n = 5 per group) and exposed to a single 15 Gy X-ray dose directed to the head. Animals were euthanized at 4, 9, 14, or 25 days after irradiation. A non-irradiated control group was euthanized on day 4. Left hemimandibles were fixed, decalcified, embedded in paraffin, and sectioned. Histological analyses were performed using hematoxylin and eosin, Masson’s trichrome, and Picrosirius Red staining under polarized light. MMP-2 expression was evaluated by immunohistochemistry and quantified by grayscale pixel intensity analysis. Irradiation induced progressive alterations in the lingual periodontal ligament. Fibroblast nuclei exhibited morphological features consistent with cellular injury beginning on day 4, with increasing changes at subsequent time points. These changes were accompanied by progressive disorganisation of collagen bundles, increased interfibrillar spaces, and loss of normal extracellular matrix architecture. Picrosirius Red analysis demonstrated a reduction in the type I-to-type III collagen ratio on days 4, 9, and 14 after irradiation, supporting the morphological changes observed with H.E and Masson’s trichrome staining. MMP-2 expression increased significantly on day 4, reached its highest level on day 9, and then progressively decreased on days 14 and 25. A single dose of X-ray irradiation induces early morphological and extracellular matrix alterations in the rat incisor periodontal ligament. The temporal association between increased MMP-2 expression, collagen fibre disorganisation, and a decreased type I-to-type III collagen ratio suggests that MMP-2 is associated with the early extracellular matrix remodelling observed on days 4 and 9 after irradiation. The subsequent decline in MMP-2 expression was accompanied by an apparent partial restoration of collagen fibre organisation on days 14 and 25.