Journal of Molecular Histology最新文献

This study aimed to determine whether low-intensity blue LED light (4 J/cm²) promotes mineralization of stem cells from the apical papilla (SCAPs) by modulating CRY1 expression and to elucidate the underlying molecular mechanisms. SCAPs identity was validated using flow cytometry. In a controlled experimental design, SCAPs were exposed to blue LED light, followed by quantitative assessment of CRY1 and osteogenic markers (Runx2, OSX, DSPP, DMP-1) via qRT-PCR, Western blotting, and osteogenic staining. To investigate the role of CRY1 in SCAPs osteogenic differentiation, CRY1 was overexpressed using lentiviral transfection. Additionally, the Wnt/β-catenin pathway was analyzed using specific inhibitors (XAV-939) to elucidate the underlying molecular mechanisms. Blue LED irradiation reduced CRY1 mRNA expression to 80% (day 7) and 45% (day 14) of control levels. CRY1 overexpression significantly increased CRY1 mRNA and protein levels (P < 0.001) but decreased ALP activity and ARS staining (P < 0.001). Blue LED treatment restored mineralization parameters to 80% of control levels. Key osteogenic genes (DMP-1, DSPP, Runx2, OSX) showed lower mRNA and protein levels in the CRY1 overexpression group compared to controls. Blue LED exposure increased these levels to 60–74% (mRNA) and 45–67% (protein) of control values. In the Wnt/β-catenin pathway, CRY1 overexpression elevated GSK-3β and reduced p-GSK-3β, β-catenin, and nuclear β-catenin levels. Blue LED treatment restored these levels to 33–54% of control values, indicating pathway activation. Inhibition of the Wnt/β-catenin pathway (using XAV-939) abolished differences in osteogenic gene expression and mineralization between CRY1 overexpression and blue LED-treated groups, confirming its critical role. Blue LED light at 4 J/cm² enhances SCAPs mineralization by modulating CRY1 expression and activating the Wnt/β-catenin pathway. These findings provide mechanistic insights into photobiomodulation (PBM) in bone regeneration and highlight its potential for tissue engineering and regenerative medicine.

We aimed to investigate whether fatty acid binding protein 4 (FABP4) knockdown can ameliorate fetal development in rats with gestational diabetes mellitus (GDM) through modulating autophagy mediated by the phosphatase and tensin homolog deleted on chromosome ten (PTEN)/serine/threonine protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway. Blank group (n = 12), GDM group (n = 12), small interfering negative control (si-NC) group (n = 12), si-FABP4 group (n = 12), and si-FABP4 + PTEN group (n = 12) were set up for the random assignment of pregnant rats. Compared to the blank group of pregnant rats, the serum FABP4 level, abortion rate, fasting insulin, homeostasis model assessment of insulin resistance index, development malformation rate and incidence rate of intrauterine growth restriction of fetal rats, as well as PTEN and Beclin-1 protein expressions and light chain 3 type II (LC3-ΙI)/LC3-Ι ratio in placental tissues rose significantly, while the phosphorylated (p)-Akt/Akt ratio, p62 protein expression, and p-mTOR/mTOR ratio in placental tissues dropped significantly in the GDM plus si-NC group (P < 0.05). FABP4 knockdown improves fetal development in GDM rats, and its mechanism of action is probably related with the inhibited autophagy by regulating the PTEN/Akt/mTOR signaling pathway.

Differentiation and proliferation of odontogenic cells are regulated by specific genes, driving the morphological changes during tooth germ development. While substantial data have been gathered on genes involved in early tooth development, knowledge about the terminal differentiation of odontogenic cells—particularly ameloblasts—remains limited. To identify molecules involved in cytodifferentiation, gene expression profiles in maxillary cap stage and root stage molar germs of rats at postnatal day 9, respectively were compared using differential display-PCR. This analysis unexpectedly revealed the upregulation of neurogranin, a brain-specific calmodulin-binding protein, at the cap stage. The expression of neurogranin at both transcriptional and translational levels progressively decreased from the early bell stage, through the crown stage, to the root stage in molar germs. Similar temporal expression patterns were observed for other calmodulin-binding proteins, including neuromodulin and PEP-19. Immunofluorescence confirmed the presence of neurogranin in the inner enamel epithelium of cap and early bell stage molar germs. Correspondingly, the expression levels of calmodulin and PKCγ mRNA displayed dynamic changes during the developmental timeline. Notably, the ablation of neurogranin in SF2 ameloblastic cells enhanced ameloblast differentiation and mineralization. These findings, for the first time, suggest that neurogranin may play a critical role in the temporospatial regulation of the differentiation of inner enamel epithelial cells into ameloblasts.

Cervical cancer (CC) remains a leading cause of cancer mortality amongst females worldwide. Some of the CC patients may experience early metastases of the primary tumor, however the underlying mechanism remains unclear. Aberrant expression of RING box protein-1 (RBX1), a subunit in the E3 ubiquitin ligase family, has been reported in several cancer types. Nevertheless, little is known regarding the role of RBX1 in the metastasis of CC patients. In this study, we examined the expression of RBX1 from 90 biopsies of CC patients, and found a significantly increased expression of RBX1 in the tumor tissues compared to the normal tissues. Notably, the abundance of RBX1 in the CC patients with metastasis was higher than their counterparts without metastasis, suggesting that RBX1 may play a significant role in the modulation of CC metastasis. Furthermore, by using Hela cells as a model of CC in vitro, we demonstrated that ectopic over-expression of RBX1 could significantly promote the migration and invasion of Hela cells, whereas knockdown of RBX1 could remarkably suppress the migration and invasion of Hela cells. Mechanistically, the regulatory effect of RBX1 on cell metastasis was associated with changes in matrix metalloproteinases (MMP3 and MMP9) and altered activity of PI3K/AKT signaling. In conclusion, this study highlighted RBX1 as a novel target that can promote the metastasis of Hela cells in vitro, which may contribute to the development of alternative therapeutic options for CC patients.

The Anaphase-Promoting Complex/Cyclosome (APC/C) is a critical regulator of cell cycle progression, with APC15 serving as an essential subunit. While the role of APC15 in mitosis is well characterized, its function during meiosis remains poorly understood. In this study, we investigated the expression, subcellular localization, and potential role of APC15 during mouse oocyte meiotic progression. Using immunofluorescence and confocal microscopy, we observed dynamic changes in APC15 localization throughout meiotic progression. Knockdown of APC15 via siRNA did not affect spindle organization, but led to meiotic arrest at metaphase I (MI) and impaired the removal of BUB3 from kinetochores, suggesting a disruption in Spindle Assembly Checkpoint (SAC) inactivation. Our results highlight the involvement of APC15 in the regulation of SAC and the transition from metaphase to anaphase in oocytes. These findings contribute to our understanding of APC15’s role in meiotic regulation and provide insights into its potential impact on maintaining chromosomal stability during oocyte maturation.

Alzheimer’s disease (AD) is a common neurological disorder primarily affecting older adults. A hallmark of this condition is the generation of reactive oxygen species (ROS), leading to increased oxidative stress and cellular damage. Treatment with a curcumin-selenium nanoemulsion has been shown to enhance behavioural performance and mitigate degenerative changes induced by aluminium chloride (AlCl₃). This nanoemulsion also reduced the activity of acetylcholinesterase (AChE) and lowered levels of key proteins, including Aβ, p53, tau, nuclear factor erythroid 2-related factor 2 (Nrf2), and tumour necrosis factor-alpha (TNF-α). Additionally, it significantly decreased nitric oxide (NO) levels in the brain while enhancing the activity of catalase (CAT) and superoxide dismutase (SOD). The study highlights the antioxidant and anti-inflammatory properties of the curcumin-selenium nanoemulsion, suggesting its potential as a therapeutic option for alleviating AD induced by AlCl₃. These results are further supported by improvements in the histological structure of the cortex and hippocampus, as well as enhanced immunohistochmical assessment of glial fibrillary acidic protein (GFAP). Cur- Se-nanoemulsion, the current drug delivery technology, may lower the amount of amyloid-β in AD rat brain and considerably ameliorate the memory deficit that improve therapy efficacy in AD lesions.

Despite numerous experimental and clinical studies in terms of novel therapeutic strategies, the treatment of acute pancreatitis (AP) is still symptomatic. In the present study, we aimed to evaluate the effects of Punica-granatum L. (pomegranate) and Aronia-melanocarpa (black chokeberry) which both have antioxidant and anti-inflammatory actions on the treatment of AP. Thirty-two rats were divided into 4 groups: Control, AP, pomegranate, and black chokeberry groups. To create AP, L-arginine was administered intraperitoneally. The rats from the pomegranate and black chokeberry groups were administered two doses of pomegranate and black chokeberry extract orally after each L-arginine injection for 7 days. Pancreatic tissues were processed for light and electron microscopic and biochemical evaluations. L-arginine administration resulted in severe necrotic AP characterized by necrosis, apoptosis, and significant increases in serum amylase, and lipase levels as well as tissue total oxidant status (TOS) levels. Degenerated organelles, apoptotic bodies, and myelinic figures were observed by electron microscopic examination. Both pomegranate and black chokeberry had beneficial effects on the treatment of AP via decreasing total histopathological damage score, apoptosis rate, and serum and tissue TOS levels. These beneficial effects of pomegranate and black chokeberry extracts may offer compelling information that may inspire and guide the development of novel therapeutic approaches for the treatment of AP.

Graphical abstract

Nigella Sativa (N. sativa) belongs to the family of Ranunculaceae and is an annual herb which is also known as black cumin or black seed. Since ancient times N. sativa has been used because of its widespread therapeutic properties which has been proven effective in respiratory, cardiovascular, gastrointestinal, inflammatory conditions and in inflammation. This study investigates the antiinflammatory, antipyretic, and analgesic properties of Nigella sativa (N. sativa) methanol extract using albino rats (n = 36). Diclofenac and paracetamol were used as standard medications in the trial, and the four concentrations of the methanolic extract (250, 500, 1000, and 2000 mg/kg) were tested for their effects on inflammation, pain, and fever. The methanolic extract of N. sativa seeds showed significant inhibition of fever (96%), inflammation (89%), and pain (85%). In addition, bioactive compounds in the seeds, including thymol, p-cymene, and limonene, were examined through in-silico studies. Ligand molecules and proteins associated with anti-inflammatory, analgesic, and antipyretic effects were 1DFN, 1A06, and 3LN0, respectively. The molecular docking results indicated significant binding interactions, with effective binding energy values corresponding to analgesic, antipyretic, and anti-inflammatory properties. Both in-silico and in-vivo results demonstrate the efficacy of N. sativa methanol extract in alleviating pain, inflammation, and fever.

Endometriosis is illustrated by the presence of ectopic endometrial cells capable of evading apoptosis outside the uterus. Apoptotic and anti-apoptotic factors in the extra uterine microenvironment can be compromised by the impairment in oxidative status. Angiotensin Converting Enzyme (ACE) Inhibitors and Nitric Oxide (NO) modulators play pivotal role in inflammation, angiogenesis, apoptosis and in abrogating oxidative imbalance. Therefore, in the current study we investigate the role of ACE inhibitor and or NO modulators in mitigating the proliferation of ectopic endometrial lesions in rat model. Sixty adult female virgin wistar rats were utilized; out of which fifteen were used as donor rats and rest forty-two were randomly divided into seven groups after surgical implantation of endometrial explants into rats (group II–VII). Histomorphometric assessment of uteri and ectopic lesions was performed by Hematoxylin and eosin (H-E) staining, followed by immunohistochemical study for Proliferating cell nuclear antigen (PCNA), Bax and Bcl-2. Oxidative stress parameters were evaluated by biochemical estimations, succeeded by immunoblotting of Poly [ADP-ribose] polymerase 1 (PARP1). Additionally, immunoblotting of Vascular endothelial growth factor (VEGF), Bax, Bcl-2 and caspase-3 was also performed. Significant decrease in the diameter of lesions with diffused staining at the extracellular spaces of stromal cells for PCNA accompanied by significant decrease in the expression of VEGF (p < 0.00001) was observed in group III. Furthermore, increased expression of Bax:Bcl-2 ratio (p < 0.001) and cleaved caspase-3 (p ≤ 0.0001) in ectopic lesions of group III was also observed. Administration of ramipril alone results in triggering oxidative stress mediated cleavage of PARP1, augmenting apoptosis in the ectopic lesions.

Metastasis is a major determinant of prognosis in gastric cancer (GC), and microRNAs (miRNAs) play crucial roles in driving the metastatic process. This study aimed to identify key miRNAs involved in GC metastasis and elucidate their underlying mechanisms. GC tissues from patients with and without metastasis were subjected to miRNA sequencing to identify differentially expressed miRNAs. Expression differences between GC and normal tissues, as well as their correlation with patient survival, were analyzed using data from The Cancer Genome Atlas and an internal cohort. miR-378d expression was measured by RT-qPCR in the internal cohort, and its association with clinicopathological features and prognosis was analyzed. Gene Set Enrichment Analysis (GSEA) was performed to investigate the potential mechanisms by which miR-378d influences GC metastasis. The findings were validated through in vitro wound healing, transwell assays, western blotting, and immunofluorescence, as well as in vivo models. MiRNA sequencing identified miR-378d as significantly downregulated in GC tissues and associated with poor prognosis. GSEA showed that miR-378d was negatively correlated with epithelial-mesenchymal transition (EMT). In vitro and in vivo experiments demonstrated that upregulation of miR-378d inhibited GC cell migration and invasion. Mechanistically, miR-378d suppressed EMT by downregulating METTL4 expression. miR-378d inhibits GC metastasis by suppressing EMT through the downregulation of METTL4, offering novel insights into the role of miRNAs in GC progression and highlighting potential therapeutic targets for intervention.