Journal of Molecular Histology最新文献

Persimmon (Diospyros kaki L.) leaves are a traditional medicinal herb used for treating many infectious and inflammatory-related conditions, including wound healing. To validate its traditional use, our study evaluates the acute toxicity and wound-healing effects of methanolic extracts of Persimmon (Diospyros kaki L.) leaves (MEPL) on excisional neck injury in rats. A uniform dorsal neck injury was created for twenty-four Sprague Dawley rats, which were randomly aligned into 4 groups and treated topically twice daily with 0.2 ml of the following: group A, rats treated with 1% CMC; group B, rats received intrasite gel; groups C and D, rats treated with MEPL (0.2 ml of 250 and 500 mg/kg, respectively). The toxicity results showed a lack of physiologic alteration or mortality in rats ingested with an oral dosage of up to 5 g/kg of MEPL. Histological screening of regenerated skin tissues revealed higher deposition of collagen, fibroblast cells, and reduced inflammatory cells in MEPL-treated rats. The topical application of MEPL led to positive modulation of Transforming Growth Factor Beta 1 (angiogenetic factor) in wound tissues, indicating increased tissue regeneration and faster wound contraction. MEPL treatment caused a significant elevation of tissue antioxidants (superoxide dismutase and catalase) and hydroxyproline (collagen) contents while reducing malondialdehyde contents. The inflammatory mediators (TNF-α and IL-6) were lower, and anti-inflammatory cytokines (interleukin 10) were higher in MEPL-treated rats than in the vehicle group. The study outcomes back up the traditional use of MEPL for wound healing, which could be linked with its phytochemicals (flavonoids and terpenoids) that require further isolation and molecular identification.

Long non-coding RNAs (lncRNAs) have emerged as pivotal regulatory molecules in cancer biology. Among these, long intergenic non-protein coding RNA 02418 (LINC02418), a recently identified lncRNA, has been linked to endometrial cancer (EC), although its function and operational mechanisms are largely unclear. The present investigation aims to elucidate the molecular mechanism through which LINC02418 influences EC pathogenesis. We employed Western blotting and quantitative real-time PCR to analyze Ras protein specific guanine nucleotide releasing factor 1 (RASGRF1) and LINC02418 expression profiles in EC tissues and cell lines. Functional analyses, including cell proliferation, migration, and invasion assays, were conducted to evaluate the impact of LINC02418 overexpression on EC cells. Xenograft mouse models were established for in vivo validation. The molecular interactions between LINC02418, miR-494-3p, and RASGRF1 were characterized using luciferase reporter and RNA pull-down assays. LINC02418 expression was significantly downregulated in EC tissues and cell lines compared to their normal counterparts. Forced expression of LINC02418 significantly suppressed EC cell proliferation, migration, and invasion in vitro. In xenograft models, LINC02418 overexpression resulted in reduced tumor burden and enhanced cell death. Mechanistically, LINC02418 enhanced RASGRF1 expression by sequestering miR-494-3p, a finding substantiated by RNA pull-down assays. The tumor-suppressive effects of LINC02418 were partially reversed by RASGRF1 silencing and miR-494-3p overexpression. Clinical analyses revealed that reduced RASGRF1 expression correlated with poor histological differentiation, advanced tumor stages, and decreased overall survival in EC patients. Our findings establish LINC02418 as a tumor suppressor that regulates EC progression through modulation of the miR-494-3p/RASGRF1 axis, highlighting its potential as a therapeutic target in EC treatment.

Genital tract infections are common causes of male infertility, and most of diagnosed men are asymptomatic. This study examined the effect of gallic acid (GA) against lipopolysaccharide (LPS)-induced testicular inflammation. Thirty-two Spraque Dawley, 2.5-3 month-old male rats were separated into four groups (n = 8). Control group; saline at 3 ml/kg, and in the GA group; GA was dissolved in saline, by gavage at 100 mg/kg for 14 days. LPS group; LPS 5 mg/kg as a single dose was given intraperitoneal on the 11th day. LPS + GA group; GA was given for 14 days and LPS 5 mg/kg on the 11th day. After 72 h of LPS injection, all samples were collected. Semen analysis, biochemical assays, histological evaluations, and immunohistochemical or Western blot analyses for nuclear factor-kappa B (NF-κB) and Prokineticin 2/prokineticin receptor 1(PK2/PKR1) pathways were performed. There was a significant decrease in body and testicular weight, sperm parameters, serum testosterone level, mean seminiferous tubule diameter, germinal epithelial thickness, and Johnsen score in the LPS group compared to control and GA groups. However, a significant increase was found in interstitial space width, percentage of abnormal sperm, NF-κB and PK2 immunoreactivities, and expression of PK2 and PKR1 proteins. In the LPS + GA group, GA administration was observed to significantly prevent these adverse effects. In conclusion, the inhibitory effects of GA on the NF-κB and PK2/PKR1 pathways not only suppressed the inflammatory response but also restored impaired sperm parameters and testicular structure. These findings indicate GA’s potential for treating testicular inflammation and protecting male reproductive health.

Laryngocutaneous fistula is one of the most important complications encountered after larynx surgery. Stem cell therapy is a promising treatment approach for the future, both without the need for surgical methods and by assisting surgical methods to close the fistula. 30 female Downey Sprague rats were divided into 5 separate groups and pharyngocutaneous fistula was created. Stem cells were administered subcutaneously to Group 1(SC-SC) after the creation of PCF, and via the tail vein to Group 2 (V-SC). The 3rd group (C) was made as a control group and while fistula formation and subsequent closure characteristics in the neck were monitored without any treatment, the rats in the 4th group (V-M/SH) were given tail vein medium/sham and the 5th group (SC-M/SH) was given as subcutaneous medium/sham group. PCF closure was significantly higher in the group given SC compared to the control group. The differences between the groups were examined by Hematoxilen Eosin staining with light microscopy and evaluated in terms of inflammatory cell infiltration, fibroblastic activity, collagen accumulation and neo-angiogenesis using the Ehrlich Hunt scale. After the experiments, all groups were stained with immunocytochemistry via TGF-β, VEGF and TNF-α antibodies. The statistical analysis was performed using the One-Way ANOVA test. When all these parameters were compared with the control group, a statistically significant difference was found.

This study investigated tempol action on genes and miRNAs related to NFκB pathway in androgen dependent or independent cell lines and in TRAMP model in the early and late-stages of cancer progression. A bioinformatic search was conducted to select the miRNAs to be measured based on the genes of interest from NFκB pathway. The miR-let-7c-5p, miR-26a-5p and miR-155-5p and five target genes (BCL2, BCL2L1, RELA, TNF, PTGS2) were chosen for RT-PCR and gene enrichment analyses. In vitro, PC-3 and LNCaP cells were exposed, respectively, to 1.0 or 2.0 mM of tempol during 48 h. In vivo, five experimental groups were evaluated regarding tempol effects in the early (CT12 and TPL12 groups) and late-stages (CT20, TPL20-I and TLP20-II) of PCa development. TPL groups were treated with 50 mg/kg or 100 mg/kg of tempol. The ventral lobe of the prostate and the plasma was collected. Tempol treatment increased miRs expression in PC-3 and LNCaP. For both cell lines, tempol decreased RELA expression. In TRAMP model, tempol increased miRNA expression in prostate for all treated groups. Tempol upregulated the miRNA expressions related to the NFκB pathway in the prostate tissue and human tumor cell lines. Their increase is mainly linked to increased cell death and delayed CaP aggressivenes. Thus, tempol’s capacity for miRNA-mediated gene silencing to decrease tissue proliferation and cell survival processes is part of its tissue mechanics.

Objective

This study aimed to elucidate the role of pyruvate dehydrogenase kinase-1 (PDK1) in cervical cancer (CC) by investigating its impact on cell proliferation, migration, and epithelial-mesenchymal transition (EMT) under hypoxic conditions.

Methods

PDK1-silenced CC cell lines were established using lentiviral shRNA technology. Cell migration and invasion were assessed through scratch and Transwell assays, respectively. Cellular activity and apoptosis-related protein expression levels were evaluated using MTT assays and western blotting. Transcriptome sequencing elucidates the regulatory pathways impacted by PDK1 silencing, and rescue experiments confirmed the underlying mechanisms. Xenograft models with nude mice were used to validate the effects of PDK1 silencing on CC progression.

Results

PDK1 silencing reduced migration, invasion, and cellular activity under hypoxic conditions while promoting apoptosis. Transcriptomic analysis revealed that PDK1 suppression downregulated the WNT4/β-catenin/FOXO1 pathway, decreasing EMT-related protein expression. Mechanistically, PDK1 enhanced β-catenin stability by inhibiting its phosphorylation through AKT-mediated GSK3β inactivation, promoting EMT and anti-apoptotic gene transcription.

Conclusions

Targeting PDK1 may provide novel therapeutic strategies specifically for CC by modulating the WNT4/β-catenin/FOXO1 pathway and associated EMT and apoptotic processes.

Facial nerve injuries lead to significant functional impairments and psychological distress for affected patients. Effective repair of these injuries remains a challenge. For longer nerve gaps, the regeneration outcomes after nerve grafting remain suboptimal due to limited sources and postoperative immune responses. Tissue engineering techniques are conventional methods for repairing peripheral nerve defects. This study explores the potential of dental pulp cells (DPCs) combined with stem cell factor (SCF) to enhance neurogenic differentiation and improve facial nerve regeneration. DPCs were isolated from rabbit dental pulp, the pluripotency of the cells was identified from three perspectives: osteogenic differentiation, adipogenic differentiation, and neurogenic differentiation. In vivo experiments involved injuring the buccal branch of the facial nerve in New Zealand white rabbits, followed by treatment with PBS, DPCs, SCF, or SCF + DPCs. Functional recovery was assessed over 12 weeks, with SCF + DPCs demonstrating the most significant improvement in whisker movement scores. Histomorphological evaluations revealed enhanced myelinated fiber density and axonal morphology in the SCF + DPCs group. RNA sequencing identified 608 differentially expressed genes, with enrichment in the TGF-β signaling pathway. In in vitro experiments, we demonstrated from multiple angles using Western blot analysis, Real-time quantitative polymerase chain reaction (QPCR) analysis, and immunofluorescence staining that SCF can promote the neurogenic differentiation of DPCs through the TGF-β1 signaling pathway. Our findings indicate that the combination of SCF and DPCs offers a promising strategy for enhancing facial nerve repair.

This study tested whether combined ceftriaxone and adipose-derived mesenchymal stem cells (ADMSCs) would defend the spinal cord against acute spinal infection (ASI) in rodent. Adult-Male-SD rats were grouped into groups 1 (SC)/2 (ASI)/3 (ASI + ceftriaxone from days 2 to 28 after ASI induction)/4 (ASI + allogenic ADMSCs from day 2 for a total of 3 doses/3 consecutive intervals by intravenous injection)/5 (ASI + combined ceftriaxone and ADMSC) and spinal cord tissues were harvested by day 28. Circulatory levels of TNF-α/IL-6 at days 7 and 28, and these two parameters in spinal fluid at day 28 were lowest in group 1, highest in group 2, significantly lower in group 5 than in groups 3/4, and significantly lower in group 3 than in group 4 (all p < 0.0001). The day-28 bacterial colony formation unit (CFU) in vertebral bone and circulatory WBC counts at the time points of days 7/14/28, and the protein expressions of upstream (TRL-2/TLR-4/MYD88/TRAF6/IKKα/IKKβ /IKBβ/p-NF-κB) and downstream (IL-1β/IL-6/TNF-α/IFN-γ/iNOS) inflammatory signalings displayed a similar pattern of inflammatory biomarkers in spinal fluid among the groups (all p < 0.0001). By day 28, the bone injury score/bone marrow density/ratio of bone volume (BV) to the bone tissue volume (TV)/ratio of bone surface (BS) to BV/ratio of BS to bone TV/trabecular number exhibited an opposite, whereas the trabecular space exhibited an alike pattern of inflammatory biomarkers among the groups (all p < 0.0001). Combined ceftriaxone and ADMSCs therapy offered an additional benefit on protecting the vertebral bone/spinal cord against ASI damage.

Autism spectrum disorder (ASD) is a group of severe neurodevelopmental disorders. This study aimed to elucidate the potential ameliorating effect of postnatal administration of MSCs-derived Exo in a rat model of ASD. Male pups were divided into control (Cont), (VPA); pups of pregnant rats injected with VPA subcutaneously (S.C.) at embryonic day (ED) 13, and (VPA + Exo); pups were intravenously (I.V.) injected with MSCs-derived Exo either at postnatal day (P) 21 (adolescent VPA + Exo) or P70 (adult VPA + Exo). They were evaluated for physiological, histopathological and immunohistochemical changes of cerebellar structure, and genetic expression of PI3k and mTOR. The VPA adult group showed increased locomotor activity and impaired social activity, and anxiety. The cerebellar histological structure was disrupted in VPA groups. VPA + Exo groups showed preservation of the normal histological structure of the cerebellum. Immunohistochemical studies revealed enhanced expression of caspase-3, GFAP, Nestin, and VEGF in VPA groups beside modifying PI3K and mTOR genetic expression. MSCs-derived Exo ameliorated most of the rat cerebellar histopathological alterations and behavioral changes. Their mitigating effect could be established through their antiapoptotic, anti-inflammatory and anti-neurogenesis effect besides modifying PI3k-mTOR signaling.