Journal of Molecular Histology最新文献

Celiac disease (CD) is a chronic autoimmune disease of the small bowel mucosa that develops because of the altered immune response to gluten, which leads to intestinal epithelium damage and villous atrophy. However, studies on regeneration of the damaged small bowel mucosa and density of intestinal stem cells (ISC) in CD persons are still scarce. We aimed to evaluate the number of small bowel mucosa cells positive for LGR5, CD138/Syndecan-1, CD71 and CXCR3 in CD and in controls with normal bowel mucosa; to find relationship between these markers and degree of small intestinal atrophy and to compare these results with our previous data about the number of CD103 + , IDO + DCs, FOXP3 + Tregs, enterovirus (EV) density and serum zonulin level. The paraffin sections of the small bowel biopsies were obtained from 26 children with CD (median age 6.5 years), and from 20 controls with normal intestinal mucosa (median age 14.2 years) and from the tissue bank of the Department of Pathology of Tartu University Hospital (from 18 participants with CD including 14 children (median age 13.2 years) and from 11subjects with normal small bowel mucosa, including one child aged 4.8 years. The number of LGR5 + , CD71 + , CD138 + , and CXCR3 + cells was evaluated using immunohistochemistry. The median number of CD138 + and CXCR3 + cells was significantly higher in the small bowel mucosa in CD compared with normal mucosa (p = 0.0002 for CD138 and p = 0.006 for CXCR3). The median number of CD71 + cells was significantly higher in normal small bowel mucosa (p = 0.005). The number of LGR5 + cells did not differ between persons with CD and those with normal small bowel mucosa (p = 0.7). A markedly increased number of CD138 + and CXCR3 + cells in the small bowel mucosa of participants with CD confirms their role in the pathogenesis of this disease. There was no expected marked difference in the density of any of the studied markers between lower or higher grade of small bowel atrophy and level of tTG-IgA in CD.

This study was to investigate the role of microRNA (miR)-330-5p derived from mesenchymal stem cells-secreted exosomes (MSCs-Exo) in cerebral ischemia-reperfusion injury (CI/RI) through targeting lysine N-methyltransferase SET domain containing 7 (SETD7). MSCs-Exo were separated and identified. MSCs-Exo were used to treat the middle cerebral artery occlusion (MCAO) rat model. By using the nerve injury score, Nissl, hematoxylin and eosin, and terminal deoxynucleotidyl transferase dUTP nick-end labeling staining, the neural function, pathological alterations, and neuronal death in MCAO rats were examined. Using an enzyme-linked immunosorbent test, tumor necrosis factor-α, interleukin (IL)-1β, and IL-6 in brain homogenate were tested. Rat brain expression levels of SETD7 and miR-330-5p were examined. Subsequently, the effects of MSCs-Exo, miR-330-5p, and SETD7 on neurological function and pathological alterations were assessed using gain and loss function tests. miR-330-5p expression was decreased and SETD7 expression was increased in the brain tissue of MCAO rats. Both MSCs-Exo and MSCs-Exo-derived miR-330-5p reduced inflammation in MCAO rats. miR-330-5p targeted SETD7, and SETD7 upregulation blocked the therapeutic effect of MSCs-Exo-derived miR-330-5p on MCAO rats. MSCs-Exo-derived miR-330-5p targets SETD7 to reduce inflammation in MCAO rats, providing a new therapeutic target for CI/RI therapy.

Traumatic brain injury (TBI) is a common neurosurgical emergency. As a macrophage in brain, microglia involves in secondary TBI injury. UCF-101, an Omi/HtrA2 inhibitor, protects against neurological disorders. This study aims to investigate the effects of UCF-101 in TBI and its mechanism. Mouse microglia cell BV2 cells were exposed to 1 µg/mL LPS to construct TBI in vitro models. Following CCK8 assay, cells were treated with LPS + UCF-101 (2, 5, 10 µM), LPS + Compound C (AMPK inhibitor, 20 µM), and LPS + UCF-101 + Compound C groups. With lactate dehydrogenase (LDH) content detection, ELISA and qRT-PCR assays were used to measure proinflammatory factors. Biomarkers of M1 (CD16/32 and iNOS) and M2 phenotypes (CD206), as well as AMPK/NF-κB pathway-related protein expression were assessed by flow cytometry, immunofluorescence, and Western blot methods. There was a decrease in M1 phenotype biomarkers and an increase in M2 phenotype biomarkers after UCF-101 treatment. UCF-101 exposure reduced TNF-α, LDH, IL-1β, IL-6, IL-8, p-NF-κB p65/NF-κB p65, and activated p-AMPK α (T172)/AMPK α (T172) expression. Importantly, further Compound C treatment counteracted these effects of UCF-101. In conclusion, UCF-101 ameliorates TBI by promoting microglia M2 polarization via AMPK/NF-κB pathways in LPS-induced BV2 cells, providing solid scientific foundation for clinical application of UCF-101 in TBI treatment.

The effect of rabies virus infection on dendritic morphology and on the expression of the MAP2 protein in Purkinje cells in the cerebellum of mice was studied. ICR mice were inoculated with rabies virus, and six days later, the mice were sacrificed, the cerebellum was removed and processed for Golgi-Cox staining or MAP2 immunohistochemistry. Infection with rabies virus altered the dendritic pattern of Purkinje cells ranged from moderate changes to accentuated retraction in the dendritic tree of some Purkinje cells. The loss of dendritic branches in the samples of mice infected with RABV was also reflected in a decrease in intersections quantified using the Sholl technique, thus suggesting dendritic pathology. Immunoreactivity to MAP2 protein in the molecular layer of the cerebellum of control mice was mainly distributed in dendrites of Purkinje cells. Some somas were faintly stained. In infected mice immunoreactivity to MAP2 was intense in somas and dendrites of Purkinje cells and in some interneurons. These results are consistent with similar findings we previously reported for the cerebral cortex and spinal cord of rabies-infected mice. But they differ from studies in other pathologies where an association between dendritic pathology and loss of MAP2 immunoreactivity has been found. Our studies in rabies contribute to suggestion that MAP2 overexpression may also be associated with alterations in dendritic morphology. MAP2 protein contributes to maintaining cytoskeleton stability. However, in rabies, increased MAP2 expression here only determined by immunohistochemistry could destabilize the cytoskeleton of dendrites. Golgi staining is considered the gold standard for the study of dendritic morphology. Its association with changes in MAP2 expression appears to provide molecular support for the concept of dendritic pathology. These results contribute to the understanding of the effect of rabies virus infection on dendritic morphology. They therefore reinforce the idea that rabies not only has a dysfunctional effect on neurons, as some authors claim, but also affects their structure.

2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is an important environmental pollutant that disturbs the immune balance of the maternal-fetal interface (MFI) and is also a common environmental factor for the formation of cleft palate (CP). Therefore, the purpose is to investigate whether TCDD can cause CP by disrupting the immune balance of the maternal-fetal interface. Fifteen C57BL/6J mice were randomly assigned to three groups: control group, TCDD group, and TCDD plus Freund’s complete adjuvant (FCA) (TCDD + FCA) group. Peripheral blood, placentas, and palatal tissues were collected for H&E, flow cytometry, and ELISA. In the TCDD group, the placental diameter, the number of placental labyrinth vessels, and the area of sponge layer cells were all significantly reduced. At embryonic day (E) 17.0, there was a significant decrease in T-helper 1 (Th1) and Th2 cells in the peripheral blood of pregnant mice. Additionally, the levels of interferon-γ (IFN-γ) and interleukin-4 (IL-4), particularly IL-4, were significantly decreased. However, after treatment with FCA, the distance between the palatal shelves was reduced, and the placental weight, the number of labyrinth vessels, and the area of the cavernous cells in the placenta also increased. The number of Th1 and Th2 cells significantly increased, returning to the levels observed in the control group, with a more pronounced increase in the number of Th2 cells. In conclusion, TCDD may induce CP by disrupting the homeostasis of the MFI. The precise mechanisms by which TCDD impacts the immune system at the MFI require further investigation.2,3,7,8-四氯二苯并-对二恶英 （TCDD） 是一种重要的环境污染物，会扰乱母胎界面 （MFI） 的免疫平衡，也是形成腭裂 （CP） 的常见环境因素。因此，目的是研究 TCDD 是否可以通过破坏母胎界面的免疫平衡来引起 CP。将 15 只 C57BL/6J 小鼠随机分为 3 组：对照组、TCDD 组和 TCDD 加弗氏完全佐剂 （FCA） （TCDD + FCA） 组。收集外周血、胎盘和腭组织用于 H&E、流式细胞术和 ELISA。TCDD 组胎盘直径、胎盘迷路血管数量和海绵层细胞面积均显著减少。在胚胎第 17.0 天 （E） 时，妊娠小鼠外周血中的 T 辅助细胞 1 （Th1） 和 Th2 细胞显著减少。此外，干扰素-γ （IFN-γ） 和白细胞介素-4 （IL-4），特别是 IL-4 的水平显着降低。然而，用 FCA 处理后，腭架之间的距离减小，胎盘重量、迷路血管的数量和胎盘中海绵状细胞的面积也增加。Th1 和 Th2 细胞的数量显著增加，恢复到对照组观察到的水平，Th2 细胞的数量增加更明显。总之，TCDD 可能通过破坏 MFI 的稳态来诱导 CP。TCDD 影响 MFI 免疫系统的确切机制需要进一步研究。

It is crucial to investigate new anti-diabetic agents and therapeutic approaches targeting molecules in potential signaling pathways for the treatment of Type 2 diabetes mellitus (T2DM). The objective of the study was to investigate the total phenolic content, antioxidant capacity, α-glucosidase, and α-amylase inhibitory activities of Bolanthus turcicus (B. turcicus), as well as their cytotoxic, anti-adipogenic, anti-diabetic, apoptotic, and anti-migration potential on adipocytes. B. turcicus samples were extracted with methanol (MeOH), ethyl acetate (EA) and aqueous (Aq) solvents. The MeOH extract had the highest phenolic content (81.14 mg GAE/g), followed by EA (74.93 mg GAE/g) and Aq (51.09 mg GAE/g). All extracts exhibited dose-dependent increases in α-glycosidase and α-amylase inhibitory activity. B. turcicus extracts showed cytotoxic effect on adipocytes with IC₅₀ values of MeOH (141.0 µg/mL) < Aq (155.3 µg/mL) < EA (199.5 µg/mL). Furthermore, B. turcicus extracts reduced lipid droplet formation and adipocyte diameter size. All extracts altered cell morphology to resemble fibroblasts. B. turcicus extracts exhibited anti-migratory effect delaying wound healing for up to 96 h. The B. turcicus extracts showed a pro-apoptotic effects on adipocytes by increasing Caspase-3 enzyme activity and the population of DAPI-positive cell with apoptotic nuclear-morphology. B. turcicus extracts upregulated the expression of the Glut-4 gene at the mRNA, protein and intracellular level in adipocytes. In conclusion, our findings indicate that B. turcicus not only exhibits strong antioxidant properties and enzyme inhibitory activities but also exerts significant anti-adipogenic and pro-apoptotic effects in adipocytes, thereby providing a comprehensive mechanism through which it may contribute to the management of T2DM. These effects highlight the potential of B. turcicus as a therapeutic agent for improving glucose homeostasis and insulin sensitivity.

Sodium valproate– a salt of valproic acid (VPA), is an anticonvulsant used in the treatment of epilepsy and a range of psychiatric conditions that include panic attacks, anxiety, post-traumatic stress, migraine and bipolar disorder etc. VPA can cause direct damage to many tissues due to accumulation of toxic metabolites. Nowadays, phytochemicals are amongst the best options for the treatment of diseases. Moringa oleifera is a popular plant in the tropics owing to its numerous pharmacological and phytochemical properties such as antiproliferative, hepatoprotective, anti-inflammatory, and cardioprotective effects. In the present study, the protective effects of Moringa ethanol extract on oxidative lung damage caused by VPA was assessed biochemically and histologically. Sprague Dawley female rats were divided into 4 groups: Control, Moringa extract (M), sodium valproate (V), and sodium valproate + Moringa extract (V + M). Doses of sodium valproate and Moringa extract (dissolved in physiological saline) were given at 500 mg/kg b.w. and 300 mg/kg b.w. for 15 days, respectively. The rats were sacrificed on the 16th day, lung tissues collected biochemical parameters (glutathione level, antioxidant enzyme activities, oxidative stress biomarker and inflammatory proteins) and histopathological findings obtained from the study indicated increased damage in lung tissue of the valproate administered group. The damage was prevented/decreased upon administration of Moringa to the valproate rats. The present findings revealed that Moringa extract had a protective and therapeutic effect against VPA induced lung damage. Moringa extract demonstrated an ameliorative effect on histopathological and biochemical parameters in valproate induced lung damage.

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Chronic oxidative stress (COS) is related to the pathophysiology of the trabecular meshwork (TM) in glaucoma. MicroRNAs (miRNAs) have a key role in the oxidative stress-mediated glaucoma. This work investigated the function of miR-126-5p in human trabecular meshwork cells (TMCs) under chronic oxidative stress (COS). The miR-126-5p inhibitor was transfected into TMCs to assess the function of miR-126-5p. The targets of miR-126-5p were predicted by bioinformatic analysis. A luciferase assay was applied to test the relationship between miR-126-5p and its target. Cell proliferation was assessed using MTT. Flow cytometry and TUNEL were used for the assessment of apoptosis. We found that the miR-126-5p level was elevated in TMCs exposed to COS. MiR-126-5p inhibitor markedly promoted TMC proliferation and inhibited the increases in apoptosis and extracellular matrix (ECM) proteins induced by COS. Heat shock protein B8 (HSPB8) was identified to be targeted by miR-126-5p. MiR-126-5p inhibitor restored the expression level of HSPB8 in TMCs under COS. Additionally, miR-126-5p depletion activated PI3K/AKT signaling in TMCs by upregulating HSPB8. HSPB8 downregulation or LY294002 treatment prevented the effects mediated by miR-126-5p inhibition on apoptosis and ECM in COS-treated TMCs. Overall, silencing miR-126-5p protects TMCs against COS-induced injury by upregulating HSPB8 to activate PI3K/AKT signaling.

Embryonic development during the preimplantation stages is highly sensitive and critically dependent on the reception of signaling cues. The precise coordination of diverse pathways and signaling factors is essential for successful embryonic progression. Even minor disruptions in these factors can result in physiological dysfunction, fetal malformations, or embryonic arrest. This issue is particularly evident in assisted reproductive technologies, such as in vitro fertilization, where embryonic arrest is frequently observed. A detailed understanding of these pathways enhances insight into the fundamental mechanisms underlying cellular processes and their contributions to embryonic development. The significance of elucidating signaling pathways and their regulatory factors in preimplantation development cannot be overstated. The application of this knowledge in laboratory settings has the potential to support strategies for modeling developmental stages and diseases, drug screening, therapeutic discovery, and reducing embryonic arrest. Furthermore, using various factors, small molecules, and pharmacological agents can enable the development or optimization of culture media for enhanced embryonic viability. While numerous pathways influence preimplantation development, this study examines several critical signaling pathways in this contex.

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Recurrent pregnancy loss (RPL) is the occurrence of two or more consecutive miscarriages before 20 weeks of gestation. Recent research has increasingly focused on the role of oxidative stress in RPL, providing insights into its underlying mechanisms and potential therapeutic targets. Oxidative stress arises from an imbalance between reactive oxygen species (ROS) production and antioxidant defenses, leading to cellular damage and inflammation. Oxidative stress has been implicated in disrupting placental blood flow, inducing apoptosis in fetal and placental cells, and exacerbating inflammatory responses, all of which can contribute to pregnancy loss. Elevated levels of ROS have been associated with compromised placental function, impaired fetal development, and increased risk of RPL. Additionally, oxidative stress can modulate maternal immune responses, potentially leading to immune-related pregnancy complications. This review synthesizes current evidence on the mechanisms by which oxidative stress contributes to RPL and highlights emerging research on potential interventions, including antioxidant therapies and lifestyle modifications. Understanding these mechanisms is crucial for developing effective preventive and therapeutic strategies to reduce the risk of RPL and improve pregnancy outcomes. Future research should focus on elucidating the specific pathways involved and exploring novel treatments aimed at mitigating oxidative damage during pregnancy.