European Journal of Histochemistry最新文献

G protein-coupled estrogen receptor 1 (GPER1) has extensively verified as a tumor regulator in various types of cancers. However, its role in esophageal cancer (EC) remains largely unclear. In this study, the expression and prognostic prediction value of GPER1 in EC was analyzed by using TCGA database and was verified in EC cells and fresh tissues. The results showed that GPER1 is decreased in EC cells and tissues, and lower GPER1 expression is associated with poor overall survival of EC patients. CCK-8 assay and flow apoptosis cytometry were applied to measure the ability of proliferation and apoptosis of EC cells with or without GPER1 overexpression. The levels of reactive oxygen species (ROS) and Fe2+ were determined by flow cytometry. Elisa and Western blotting were employed to measure the markers of ferroptosis and cyclic adenosine monophosphate (cAMP) pathway. The results of in vitro experiments indicated that overexpression of GPER1 caused decreased proliferation, increased cell apoptosis, ROS generation, Fe2+ content and acyl-CoA synthetase long-chain family member 4 (ACSL4) expression, while decreased glutathione peroxidase 4 (GPX4) expression. Notably, the cAMP/PKA inhibitor H89 significantly reversed the ferroptotic effects induced by GPER1, indicating the essential role of the cAMP pathway in this process. The weight and volumes of tumors were measured and Ki-67 and H&E staining were conducted to analyze the effect of GPER1 in vivo. The results of in vivo experiments indicated that overexpression of GPER1 resulted in restricted tumor growth, reduced Ki-67 expression and increased cell death. In conclusion, the expression of GPER1 is reduced in EC. Overexpression of GPER1 enhances ferroptosis in EC, primarily through activation of the cAMP signaling pathway.

Ozone (O3) is an oxidizing natural gas widely applied as adjunctive therapeutic treatment for a variety of pathological conditions. Currently, O3-based therapies rely on the low-dose concept i.e., the administration of low O3 concentrations able to induce a mild oxidative stress stimulating antioxidant and anti-inflammatory response without causing cell damage. In addition, low O3 concentrations are thought to activate cellular and molecular mechanisms responsible for analgesic and regenerative effects. Due to these properties, in the last decade interest has arisen in the fields of orthopedics and regenerative medicine on the potential of O3 to counteract joint diseases involving cartilage degeneration. In this pilot study, we have explored the anti-degenerative potential of O3 on knee articular cartilage explanted from a healthy adult rabbit and maintained in vitro. Light and transmission electron microscopy were used to monitor chondrocyte and extracellular matrix features of cartilage samples undergoing O3 treatment every three days for two weeks. Results demonstrated that low O3 concentrations act on chondrocytes and the molecular components of the extracellular matrix of articular cartilage explants, significantly improving their preservation under in vitro conditions, likely by promoting both protective and pro-regenerative pathways. This opens promising perspectives for further investigations on the therapeutic potential of O3 for the treatment of cartilage degeneration not only as painkilling and anti-inflammatory agent but also as a cartilage regenerative agent.

We identified vagal innervation in the pharyngeal tooth and jawbones of Nile tilapia through macroscopic observations and immunohistochemistry. We also revealed the apposition of the nerve and osteoclasts in the pharyngeal jaw, suggesting the possibility of neuronal regulation for bone remodeling. However, the central projection from the vagal nerve, which innervates the pharyngeal jaws, remains unknown. To determine the projection of the vagus nerve in the brain, we applied carbocyanine dye (DiI) into the vagus nerve, revealing DiI-labeled neurons in the caudal vagal ganglion. The labeled fibers of the neurons were then traced to the vagal lobe, revealing that they branched and ran dorsally before terminating in a band-like pattern. Meanwhile, the labeled fibers running ventral to the vagal lobe were directed toward the dorsal motor nucleus of the vagus and did not have a definite terminal structure. The vagus nerve innervates the pharyngeal jaw, mainly projects to the vagal lobe, where it receives gustatory information. Pharyngeal tooth-derived sensory information might occur during occlusion and be processed precisely for determining the regurgitation and swallowing of prey.

Cold shock domain (CSD) proteins, such as YB-1, play a crucial role in the regulation of transcription, mRNA stability, and translation. Consequently, YB-1 is implicated in processes such as cell differentiation, oncogenesis and oxidative stress response. The development of the eye is a complex process that involves the differentiation of numerous highly specialized cell types. We hypothesized that YB-1 is involved in both eye development and stress defense mechanisms. As an initial step, we investigated the expression of YB-1 during the embryology of the mouse eye. YB-1 mRNA could be detected by RT-PCR and sequencing the PCR product in retinal tissue of adult mice. To elucidate the expression pattern of YB-1 protein during mouse eye development, we analyzed its expression in the developing mouse eye at embryonic day 13 (E13), E15, E18 and postnatal day 14 (P14) using immunohistochemistry. Expression of the YB-1 protein was detected in all retinal cells, as well as in the corneal and lens epithelial cells, throughout all stages of eye development examined. These findings suggest that YB-1 could have a significant role in the eye, potentially related to development and differentiation.

Metabolism-associated fatty liver disease (MAFLD) is a liver disease characterized by hepatic steatosis and excessive accumulation of lipids, with a high global incidence, especially in populations with obesity, diabetes and metabolic syndrome (MetS). As an important B vitamin, folate (FA) is stored mainly in the liver where it regulates oxidative stress, chronic inflammation and lipid metabolism. However, its regulatory role and mechanism of action in MAFLD are still poorly understood. Therefore, this study was conducted to investigate the regulatory effect of FA on MAFLD. The MAFLD rat model was induced by a high-fat diet (HFD), and HepG2 cells were treated with 0.3 mM palmitic acid (PA) for 24 h to establish a cell model. The expression of relevant genes and proteins was detected by RT-qPCR and Western blotting. Injury to HepG2 cells and rat liver tissues was evaluated via hematoxylin and eosin staining, Oil red O staining, ELISA and CCK-8 assay. FA treatment inhibited body weight gain in rats and reduced the levels of liver injury indicators (aspartate and alanine aminotransferase, and Alkaline phosphatase), blood lipids (total cholesterol, triglycerides and free fatty acids) and inflammatory cytokines (TNF-α, IL-6, and IL-1β), reducing lipid accumulation and pathological damage in the liver and ultimately alleviating the progression of MAFLD. Moreover, FA treatment promoted the expression of the autophagy-related protein LC3 II/I, inhibited the expression of p62, and increased the formation of autophagosomes, thereby alleviating PA-induced damage to HepG2 cells. Furthermore, NRF2 expression is downregulated in MAFLD and can be upregulated by FA treatment. Further examination revealed that knocking down NRF2 could partially attenuate the inhibitory effect of FA on PA-induced HepG2 cell injury. In conclusion, FA activates autophagy by promoting the expression of NRF2, thereby alleviating the development of MAFLD.

This corrects the article published in the European Journal of Histochemistry 2013;57:e40.

To elucidate the proteins associated with cochlear development and auditory formation from a histomorphological point of view, this study examined the spatio-temporal expression pattern of nestin, parvalbumin, and otoferlin in the mouse cochlea from embryonic day 17 (E17) to postnatal day 28 (P28) using immunofluorescence. Our findings revealed that nestin was broadly expressed in developing otic mesenchyme cells beneath the basilar membrane, medial to the greater epithelial ridge, and adjacent to the developing stria vascularis during late embryonic stages (E17 and E18.5). From P1 to the onset of hearing (P14), nestin was primarily expressed in fibrocytes derived from otic mesenchyme cells in the spiral ligament and spiral limbus, as well as in tympanic border cells. Dual immunofluorescence staining of nestin with Isolectin B4 (IB4), a specific vascular endothelial marker, showed the location of nestin in the blood vessels within the cochlear lateral wall. Notably, in adults (P28), nestin expression was downregulated in the fibrocytes of the spiral ligament and spiral limbus but persisted in the tympanic border cells. Parvalbumin immunolabeling was consistently observed in spiral ganglion neurons (SGNs) and inner hair cells (IHCs) from E17 through adulthood. By P1, parvalbumin expression extended to all three rows of outer hair cells (OHCs) and persisted into adulthood. Transient parvalbumin expression was also noted in afferent nerve fibers innervating the IHCs during early postnatal stages. Otoferlin labeling was predominantly detected in the cytoplasm of IHCs, with limited temporal expression in OHCs from P6 to P10. Taken together, these results illustrated the dynamic expression of nestin, parvalbumin and otoferlin during cochlear development and suggested their important function in cochlear development.

Myelofibrosis (MF) is a rare chronic hematological disorder, within the family of myeloproliferative neoplasms. The MF patients present clinical abnormalities such as anemia, and thrombosis, as well as alterations in the bone marrow (BM) microenvironment, an increased number of megakaryocytes (MKs), most of which are found in emperipolesis with neutrophils. In MF, the MKs emperipolesis is induced by an altered MK secretome, containing increased levels of pro-inflammatory cytokines, proteins, and growth factors such as interleukin-8 (IL-8) and P-selectin (P-sel). These, allow the altered cell-to-cell interactions and cause the transforming growth factor-β (TGF-β) to be released into the BM microenvironment. This fibrogenic cytokine contributes to BM fibrosis and disease progression. Emperipolesis has already been identified as a pathobiological event that contributes to MF and it is widely recognized in the most advanced stages of the disease. In this study, we evaluated the role of P-sel in BM alterations associated with emperipolesis in the Gata1low mouse model of MF. Our data show that emperipolesis is driven by P-sel. Genetic ablation of P-sel rescued the BM microenvironment, by decreasing fibrosis, suggesting that pharmacological targeting of P-sel could contribute to reduce the BM dysfunction and disease progression.

The bone marrow (BM) niche plays a pivotal role in regulating the fate of hematopoietic stem cells (HSCs), and its integrity changes significantly during aging and in rare hematological disease, as in myelofibrosis (MF). In this study, we investigated how the localization and dynamics of HSCs are influenced under physiological and pathological conditions by a newly identified by HSC-supportive megakaryocytes (MKs) subpopulation. Using huCD34tTA/TetO-H2BGFP reporter mice, we analyzed HSCs distribution within the BM and quantified nuclear green fluorescent protein (GFP) intensity to assess the repopulating potential of aged controls and mutated Gata1low mice for MF. In the control group of aged mice, cells with high levels of GFP are clustered, and adjacent to cells morphologically identifiable as supportive MKs. These clusters displayed homogeneous GFP intensity, indicating that HSCs with similar functional properties tend to co-localize in proximity to supportive MKs. By contrast, in aged huCD34/TET/Gata1low mice, GFP cells were predominantly isolated and showed reduced fluorescence intensity. Although the frequency of MKs with a supportive phenotype was increased in MF mice, analyses of GFP revealed that the ability of these MKs to maintain the HSCs in their niche was significantly impaired. Our results provide new insights on the maladaptive remodeling of the BM niche. They highlight the supportive role of MKs as potential key regulators of HSCs homeostasis. Despite their numerical expansion in MF, these cells are functionally compromised, thereby contributing to altered HSCs localization, mobilization, and to hematopoietic failure.

The functional state of placental mesenchymal stem cells (PMSCs) plays a critical role in maintaining maternal-fetal interface homeostasis during the pathogenesis of preeclampsia (PE). Given the limitations associated with direct stem cell transplantation, this study aimed to investigate the therapeutic potential of PMSC-derived exosomes and their carried miR-146a-5p. Clinical sample analysis revealed a significant downregulation of miR-146a-5p in placental tissues from PE patients, accompanied by impaired proliferation, migration, and angiogenic dysfunction of PMSCs. In an in vitro model, exosome intervention effectively reversed hypoxia-induced trophoblast cell apoptosis and enhanced migratory capacity. Furthermore, it promoted macrophage polarization towards the anti-inflammatory M2 phenotype and markedly improved the inflammatory cytokine secretion profile. In a PE mouse model, exosome treatment reduced maternal blood pressure and proteinuria levels, alleviated fetal growth restriction, and up-regulated the expression of M2 macrophage markers in placental tissue. Mechanistically, miR-146a-5p targeted TRAF6 to suppress NF-κB pathway activation, an effect that could be reversed by specific inhibitors. This study is the first to demonstrate that PMSC-derived exosomes, via the miR-146a-5p/TRAF6 axis, concurrently ameliorate trophoblast dysfunction and correct macrophage polarization imbalance. The efficient intercellular delivery of miR-146a-5p by exosomes underscores their potential as a novel targeted therapeutic strategy for PE.