Journal of Molecular Histology最新文献

Recently, microRNAs (miRNAs) have been found to mediate the development of diabetic kidney disease (DKD) by regulating podocyte injury. The aim of this study was to investigate the influence of miR-504-3p on high glucose (HG)-treated mouse renal podocytes (MPC5) and its potential regulatory mechanisms. First, a DKD cell model was established. Next, RT-qPCR was performed to measure miR-504-3p and HNF1 Homeobox B (HNF1B) expression levels. Additionally, the proliferation and apoptosis of MPC5 cells were assessed using CCK-8 assay and Flow cytometry, respectively. The protein expression levels of cell fibrotic markers, podocyte injury marker, epithelial-mesenchymal transition (EMT) markers and HNF1B were measured by Western Blotting. ROS, MDA, SOD and GSH kits were used to assess oxidative stress levels. Furthermore, the interplay between miR-504-3p and HNF1B was confirmed by luciferase reporter experiments. The miR-504-3p expression was significantly upregulated in GEO database (GSE161884) and in HG-induced MPC5 cells. The results revealed that HG treatment decreased MPC5 cell proliferation, promoted cell apoptosis and fibrosis, and ultimately led to podocyte injury. However, miR-504-3p knockdown could reverse these phenotypes and reduce podocyte injury. Moreover, online database screening combined with dual luciferase reporter assay confirmed HNF1B as a specific target of miR-504-3p. Finally, overexpression of HNF1B mitigated the proliferation inhibition and apoptosis promotion induced by oxidative stress and inhibited EMT-mediated cell fibrosis, thereby counteracting the effects of miR-504-3p on podocyte injury under HG treatment. In summary, our data indicate that miR-504-3p regulates HG-induced podocyte injury by sponging HNF1B, providing a new direction for the treatment of DKD.

Background

As a member of the interleukin-6 family, ciliary neurotrophic factor (CNTF) regulates inflammation, oxidative stress, and other processes to exhibit neurotrophic and differentiating effects over cells in the central nervous system. It has not yet been documented, therefore, if CNTF influences the cardiac remodeling brought on by myocardial infarction (MI). The purpose of the current investigation was to identify the function and underlying mechanisms of CNTF in cardiac remodeling brought on by MI.

Methods

Using an adeno-associated virus 9 (AAV9) system and tail vein injection, we overexpressed CNTF in the hearts. To create a model of MI, C57BL/6 mice underwent left anterior descending (LAD) ligation. The following techniques were employed to assess the impact of CNTF overexpression and the underlying mechanisms: quantitative real-time PCR, western blotting, histological analysis, immunofluorescence and immunohistochemistry analysis, and echocardiography. We used H9c2 cells to confirm CNTF’s in vitro effects.

Results

In MI mice, overexpression of CNTF prevents cardiac hypertrophy and cardiac fibrosis. Furthermore, oxidative stress and ferroptosis in response to MI damage were markedly reduced by CNTF overexpression. Mechanistically, overexpression of CNTF in both in vivo and in vitro markedly enhanced PI3K/Akt signaling. However, blocking this pathway effectively negated the beneficial impact of CNTF overexpression.

Conclusions

Our research indicates that via initiating the PI3K/Akt signaling pathway, CNTF controls myocardial dysfunction, oxidative stress, and ferroptosis in MI-induced cardiac remodeling. CNTF may have therapeutic potential in treating MI-induced cardiac remodeling.

Sleep is an essential health requirement; human body needs sufficient amount and quality of sleep to ensure its health. Sleep disturbance led to deterioration in renal functions. This study aimed to asses effect of sleep disturbance on nephrin protein in renal corpuscle. A sample of thirty adult male albino rats, subjected to sleep disturbance by light, divided into three groups; control group with normal sleep rhythm of 12–12 h dark- light phases, group A: subjected to interruption of sleep by light at three intervals, group B: rats were exposed to a reduction in sleep time by continuous light stimulation for 7 h. Animals were sacrificed by euthanasia, their kidneys were dissected and prepared for paraffin, sections stained for Nephrin protein, and the immunohistochemical intensity was quantified by Aperio Image Scope analysis software. This study showed variations in the effect of sleep disturbance patterns by light exposure on nephrin protein expression in renal corpuscles; in the control group a strong patchy distribution of Nephrine in the peripheral region of the glomerulus, group A showed a significant reduction compared to the control group, and group B a weak expression of nephrin protein in the glomerulus, with significant changes between group B and group A, but no significant changes between group B and control. These changes reflect that sleep disturbance affects the structural integrity of the slit diaphragm and nephrin protein expression, which is considered a novel protein for the slit diaphragm structural integrity, and a sign of podocyte injury.

The Sirtuin-1 (Sirt1) gene has been reported to be closely associated with the progression of multiple diseases, but its role in regulating osteomyelitis (OM) pathogenesis has not been explored. The murine long bone-derived osteocyte-like MLO-Y4 cells and osteoblast-like MC3T3-E1 cells were exposed to Staphylococcal protein A (SpA) treatment to establish the in vitro OM models. The expression levels of Osteoblast-specific genes (OCN, OPN and RUNX2), osteoclastic genes (CTSK, MMP9 and ACP5) and the FOXO pathway-related proteins (FOXO1, p-FOXO1, FOXO3 and p-FOXO3) were detected by performing Real-Time qPCR and Western Blot analysis. Osteoblastic differentiation of the cells were evaluated by using the alizarin red S staining assay and TRAP staining assay, and membrane potential and superoxide production were measured to evaluate the mitochondrial functions of the cells. SpA treatment significantly suppressed osteogenic differentiation and induced mitochondrial dysfunction in MLO-Y4 and MC3T3-E1 cells, and promoting osteoclastogenesis in RAW264.7 cells, suggesting that the in vitro OM models were successfully established. Of note, SpA decreased the expression levels of Sirt1 in the OM cells, and SpA-induced detrimental effects on the OM cells were all reversed by overexpressing Sirt1. Mechanistically, Sirt1-overexpression increased the levels of phosphorylated FOXO-related proteins (p-FOXO1 and p-FOXO3) to activate the FOXO signal pathway and ameliorated OM progression in SpA-treated cells. Collectively, it was revealed in the present study that overexpression of Sirt1 activated the FOXO signal pathway to ameliorate SpA-induced detrimental effects in the OM cells, and Sirt1 could be potentially used as therapeutic agent for OM in clinic.

This study analyzes the morphology of melanomacrophage centers (MMCs) in the kidney, liver, and spleen of Gymnotus carapo across two different environmental settings with varying degrees of anthropogenic impact, aiming to link habitat conditions to fish health and validate MMCs as bioindicators for environmental monitoring. A total of 28 specimens, captured and humanely sacrificed under anesthesia, were processed using conventional histological techniques. Morphometric measurements were recorded, and tissue samples were examined on an Olympus BX 41 microscope, selecting 10 random 10x fields per organ to count MMCs. Macroscopic and histological examination of the liver, kidney, and spleen highlighted MMCs distribution patterns and notable differences across sex and age groups. Adult and juvenile males showed major MMCs counts in the kidney and spleen compared to females, while liver samples from both sexes contained only pigmented cells without significant MMCs formation. Differences in MMCs quantity and structure were also observed between environments: natural habitats displayed lower MMCs counts in both sexes compared to samples from locations with higher anthropogenic impact, where MMCs frequency and pigmentation intensity were notably elevated. These findings suggest that MMCs variations in G. carapo may reflect environmental conditions and stressors, supporting the potential application of MMCs as health indicators for aquatic organisms and ecosystem monitoring. This work provides comprehensive morphometric and histological data on MMCs distribution in G. carapo, emphasizing its potential as a bioindicator species, particularly valuable in regions with varying pollution levels.

The latest research shows that the imbalance of reactive oxygen species (ROS) leads to oxidative stress-induced osteoblast apoptosis, which is an important factor in the development of osteoporosis. Methyltransferase like 3 (METTL3) is the most widely known methyltransferase, which has a marked effect on the cells of oxidative stress reaction. However, the precise mechanism through which METTL3 mediates oxidative stress-induced osteoblast apoptosis remains uncleared. An ovariectomized (OVX) rat model was established and histochemical staining were used to evaluate bone mass and the expression of METTL3. The oxidative stress state of bone tissue and the expression of METTL3 were detected by RT-PCR. The reactive oxygen species (ROS) levels were detected by DCFH-DA staining. Cell death and apoptosis were detected by CCK8, Hoechst PI double dyeing and TUNEL staining. The mitochondrial membrane potential was detected by JC-1 fluorescent staining. The expression of N6-methyladenosine, the protein levels of cell apoptosis and Wnt/β-catenin signal were detected by RT-PCR and western blot. We demonstrated that METTL3 was highly expressed in OVX-induced osteoporosis, and it inhibited oxidative stress-induced apoptosis of MC3T3-E1 cells by downregulating the ROS-mediated activation of the Wnt/β-catenin signaling pathway in osteoblasts. In addition, under oxidative stress, ROS accumulation further inhibited METTL3 expression and activated the Wnt/β-catenin signaling pathway, which ultimately led to apoptosis of MC3T3-E1 cells. This study investigated the important role of METTL3 in oxidative stress-induced osteoblast apoptosis. It may be a new therapeutic target for osteoporosis from the perspective of oxidative stress.

Wound healing is a complex process involving molecular and structural interactions. Transforming growth factor β (TGF-β), the renin-angiotensin system (RAS), and other important mediators play a central role on wound healing process. This study examined the dynamics of healing in healthy, hypertensive, and diabetic rats treated with or without Losartan, focusing on healing rate, scar characteristics, and molecular modulation. Macroscopic and microscopic analyses revealed delayed healing and reduced collagen deposition in diabetic and hypertensive rats compared with normoglycemic controls. Losartan affected healing by regulating TGF-β expression and collagen organization. In the groups of hypertensive and diabetic rats treated with losartan, healing aesthetics improved by less collagen deposition and consequently minor chances to fibrosis development, probably due to lower TGFβ and SMADs expression. Diabetic rats showed reduced skin and collagen fiber thickness, whereas hypertensive rats showed better healing under Losartan treatment (LT). These results demonstrate the complex interactions between LT, diabetes and hypertension on important fibrotic and inflammatory pathways. Although LT successfully reduces TGF-β expression and classical SMAD signaling in hypertensive settings, its minor effect in diabetes conditions indicate the necessity of supplemental treatments that target mechanisms unique to hyperglycemia, such as glycation end products or oxidative stress inhibitors. To improve treatment outcomes for individuals with diabetes and hypertension comorbidities, future studies should investigate the combination of multi-pathway modulators.

A significant correlation between type 2 diabetes mellitus (T2DM) and mood has been reported. However, the specific mechanism of mood’s role in T2DM is unclear. This study aims to discover mood-related biomarkers in T2DM and further elucidate their underlying molecular mechanisms. The GSE81965 and GSE55650 datasets were sourced from public databases, and mood-related genes (MRGs) were retrieved from previous literature. Initially, differentially expressed MRGs (DE-MRGs) were obtained by combining differential expression analysis and weighted gene co-expression network analysis (WGCNA). Subsequently, the DE-MRGs were incorporated into the LASSO and SVM to identify diagnostic biomarkers for T2DM. Four machine learning methods were utilized to construct the diagnostic models in T2DM, and the model with the optimal algorithm was screened. Further, based on biomarkers, functional enrichment, immune infiltration, and regulatory network analyses were conducted to excavate deeper into the pathogenesis of T2DM. In vivo experiments were used to validate the expression of the biomarkers. A total of 23 DE-MRGs were identified by overlapping 723 DEGs and 64 key modules, and there were strong positive correlations between these DE-MRGs. Afterward, KCTD16, SLC8A1, RAB11FIP1, and RASGEF1B were identified as biomarkers associated with mood in T2DM, and they had favorable diagnostic performance. Meanwhile, the RF diagnostic model constructed based on biomarkers was performed optimally and had high diagnostic accuracy for T2DM patients. Animal experiments indicated that expression levels of SLC8A1, RAB11FIP1, and RASGEF1B in T2DM were consistent with the microarray results. In conclusion, KCTD16, SLC8A1, RAB11FIP1, and RASGEF1B were identified as biomarkers related to mood in T2DM.

Neuroblastoma (NB) is a prevalent extracranial malignant neuroendocrine tumor in children, originating from the sympathetic nervous system. This study aims to investigate new therapeutic targets for NB. The differentially expressed genes were screened by analyzing the GSE35133 and GSE90689 datasets. Hub genes were identified by constructing a protein–protein interaction network. The diagnostic value of the hub genes was assessed through the analysis of receiver operating characteristic (ROC) curves and the expression, prognosis, and immune infiltration of IFI27 in pan-cancer were analyzed on the online website Sangerbox. The hub gene expression levels were validated by performing real-time reverse transcriptase-polymerase chain reaction. The functions of IFI27 in NB were investigated by Cell Counting Kit-8, 5-ethynyl-2'-deoxyuridine, wound healing, and Transwell assays. Six candidate genes (IFI27, TNFSF10, IFI44, DDX58, HIST1H1C, and HIST1H1E) were identified as potential diagnostic biomarkers for NB. The expression levels of IFI27, TNFSF10, IFI44, and DDX58 were significantly decreased, while HIST1H1C and HIST1H1E were elevated. Notably, IFI27 displayed correlations with prognosis and immune infiltration in multiple tumors. In vitro, functional assays demonstrated that the knockdown of IFI27 promoted the proliferation, migration, and invasion of U251 cells. Conversely, in SK-N-AS cells, IFI27 overexpression inhibited cell proliferation, migration, and invasion. IFI27 was lowly expressed in NB and participated in the progression of NB, which provides a new insight into the pathogenic mechanism and novel therapeutic strategy for NB.

Alzheimer disease (AD) is a neurodegenerative condition. Thymoquinone (TQ) is a natural compound that possesses beneficial biological effects on the brain. The present study evaluates the protective impact of TQ on the cerebellum in rats with AlCl3-induced Alzheimer's disease. Four groups were utilized. Control: 20 rats that were subdivided into two subgroups. Ia: received distilled water for 4 weeks. Ib: received corn oil via oral gavage (1 ml/kg daily) for 4 weeks. TQ group: 10 rats received TQ in corn oil via oral gavage (20 mg/kg daily) for 4 weeks. AD group:10 rats received AlCl3 in distilled water via oral gavage (300 mg/kg daily) for 4 weeks. AD & TQ group: 10 rats received both AlCl3 & TQ for 4 weeks. The grip period in the rotarod test decreased, escape latency in first three days and the entry latency period to the quadrant with the removed escape platform in the Morris water maze test increased in AD group, but when TQ was administered concurrently, there was a noteworthy improvement. Meanwhile, when compared to AD group, the addition of TQ showed a significant decrease (P < 0.05) in levels of malondialdehyde (MDA) and nitric oxide (NO), associated with a significant increase (P < 0.05) in reduced glutathione (GSH) level. Furthermore, AD & TQ group exhibited substantial preservation of the cerebellum's histological structure, the Purkinje cells number and transverse diameter showed a high significant increase (P < 0.001) and a significant increase (P < 0.05), respectively in comparison to the AD group. Using TQ showed improvement in behavioral tests, biochemical and histological findings. Thus, TQ might have therapeutic effects on Alzheimer’s disease.