Journal of Molecular Histology最新文献

Mechanistic studies have been suggested that adverse effect of bleomycin is attributed to formation of free radicals, mitochondria damages, oxidative stress and inflammation in lung tissue. Mitochondria act as central regulators in the oxidative stress and inflammatory responses in lung tissue, then it can be a promising approach for management bleomycin-induced pneumotoxicity. In the current study, we aim to investigated the injection of exogenous mitochondria into blood as one of the most promising pharmacological approaches to reduce bleomycin-induced lung toxicity in rats. Rats were divided into 4 groups as control, bleomycin (5 mg/kg), bleomycin + mitochondria (250 µg/kg), and mitochondria (250 µg/kg) alone. After 2 weeks, the survival rate, weight changes of animals, wet/dry ratio of lung tissue, alterations of histopathology, hydroxyproline content, oxidative stress and mitochondrial biomarkers were determined. Except the survival rate, weight changes of animals and wet/dry ratio of lung tissue, administration of bleomycin resulted in significant alteration in GSH content, MDA level, hydroxyproline amount, collapse of mitochondrial membrane potential (MMP), reduction of succinate dehydrogenases (SDH) activity and histopathological abnormality in comparison with control group. While exogenous mitochondria could inhibit GSH depletion, reduce production of MDA, improve the activity of SDH, prevent loss of MMP and histopathological abnormality. To the best of our knowledge, our data provides the first direct experimental evidence that injection of exogenous mitochondria into blood is capable of ameliorating bleomycin-induced lung toxicity in rats. These findings support that mitochondrial transplantation can be a promising therapeutic strategy for bleomycin-associated mitochondrial dysfunction and lung damage.

Acute myocardial infarction (AMI) is a leading cause of heart failure, often accompanied by myocardial fibrosis (MF), characterized by excessive extracellular matrix accumulation. Endothelial-to-mesenchymal transition (EndMT) plays a key role in MF progression post-AMI. Neuregulin-1 (NRG-1), a growth factor with cardioprotective properties, has emerged as a potential therapeutic target. Tongxinluo (TXL), a traditional Chinese medicine, mitigates MF by upregulating NRG-1. This study elucidates the mechanisms underlying the protective effects of NRG-1 and TXL against MF following AMI. Left anterior descending artery ligation established a model for mice with AMI. Adeno-associated virus was used to modulate NRG-1 expression in the myocardium. Echocardiography assessed cardiac function, and histological staining was used to evaluate MF. Expression levels of markers for myofibroblasts (α-SMA, FSP-1) and endothelial cells (CD31, VE-cadherin) were analysed to investigate EndMT. The involvement of the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signalling pathway in NRG-1’s protective mechanism was validated using biochemical methods. Tongxinluo was administered to mice with AMI via gavage for 4 weeks, and its effects on cardiac function, MF and EndMT were assessed. Overexpression of NRG-1 in mice with AMI ameliorated cardiac dysfunction and reduced interstitial and perivascular fibrosis, whereas NRG-1 deficiency exacerbated these effects. NRG-1 protected against EndMT, as evidenced by changes in myofibroblast and endothelial cell markers. The PI3K/AKT signalling pathway was involved in NRG-1’s protective mechanism against MF. The administration of TXL to mice with AMI improved cardiac function and reduced MF by activating NRG-1. Furthermore, TXL inhibited EndMT post-AMI through the NRG-1/PI3K/AKT pathway. NRG-1 and TXL protect against MF post-AMI by mitigating EndMT through the PI3K/AKT pathway. These findings suggest that targeting NRG-1 or using TXL may be promising therapeutic strategies for MF following AMI.

Intrahepatic cholestasis of pregnancy (ICP) is a liver disease that manifests predominantly in the later stages of pregnancy. The primary treatment currently involves the use of ursodeoxycholic acid (UDCA). In this study, the therapeutic effectiveness of 4-phenylbutyric acid (4-PBA) in the treatment of ICP, as well as the potential mechanisms involved, are investigated to offer new references for clinical treatment decisions using ICP model. The therapeutic effect of 4-PBA on ICP was evaluated by drug therapy on ICP cells and animal models, and corresponding fluorescence immunoassay, electron microscope, WB and other experiments. In addition, the cells and animals treated with GPR30 inhibitor were treated to investigate whether 4-PBA promoted the expression of bile salt output pump (BSEP) protein through GPR30-PI3K pathway, thereby promoting bile acid excretion. Administration of 4-PBA significantly reduced the incidence of stillbirth associated with ICP. 4-PBA was effective in decreasing serum bile acid levels, reducing the activation of the GPR30-PI3K pathway, and increasing the expression of BSEP protein in hepatocytes. 4-PBA was effective in reducing bile acid levels and significantly improving fetal outcomes associated with ICP. The potential mechanism involves the promotion of BSEP localization and expression in the hepatocytes’ microvilli structures via the GPR30-PI3K pathway.

Chemotherapy remains the primary therapeutic strategy for most tumors, particularly those at advanced stages with distant metastases and resistance to molecularly targeted therapy or immunotherapy. There are many manifestations of chemotherapy-induced gastrointestinal toxicity (CIGT), including chemotherapy-induced diarrhea (CID) and chemotherapy-induced constipation (CIC). Although the World Health Organisation and the International Association Against Cancer have different grading criteria and strategies for the prevention and treatment of CIGT, there are still many unanswered questions that need to be clarified. This review critically describes pathological mechanisms and clinical research, analyzing the variability in diagnostic criteria and the absence of standardization in grading severity. We identify a critical gap in understanding the molecular underpinnings of CID and CIC and suggest targeted areas for future research, including developing personalized treatment approaches based on genetic profiling. The findings suggest a comprehensive treatment approach combining pharmacological and non-pharmacological strategies to enhance life quality and treatment adherence. This review will offer a comprehensive bird-eye of pathophysiological mechanisms, clinical manifestations, and therapeutic strategies of CIGT, thereby enriching accessible references to clinicians, and helping them to prevent and control CID and CIC.

Vincamine, a monoterpenoid alkaloid, and an active constituent of plant Catharanthus roseus Linn, has been proclaimed for antioxidant and anti-inflammatory activities. This study was designed to evaluate the gastroprotective activity of Vincamine by ameliorating gastric ulcer in BALB/c mice. The study was also designed to find the possible mechanism of gastric protection by exploring the impact of Vincamine on gastric pH, acidic content, observing histopathology and molecular expression of inflammatory mediators like Interleukin- β (IL-1β), Interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF- α) and oxidative stress markers in the gastric tissue. A total number of 36 BALB/c mice were divided into 6 groups mainly normal control (NC) treated with normal saline, disease control (DC) treated with high dose of absolute ethanol (5ml/kg) while treatment groups involved pretreatment with low- dose Vincamine (VLD) at 10mg/kg body weight, medium-dose vincamine (VMD) at 20mg/kg body weight and high- dose vincamine (VHD) at 40mg/kg body weight before ethanol high dose administration and reference drug control, omeprazole (OMT) at the dose of 20 mg/kg body weight. Molecular expression levels of mRNA expressions of inflammatory cytokines like IL-1β, IL-6 and TNF- α were evaluated by using reverse transcription real time polymerase chain reaction method (RT-PCR). Pre-treatment of DC group with low (VLD), medium (VMD) and high doses (VHD) of vincamine improved gastric ulcer score and ameliorated histopathological parameter such as, infiltration of inflammatory cells, edema, fibrinoid necrosis, hemorrhage, and erosion score when compared to DC group. Induction of gastric model significantly increased (all P < 0.05) the mRNA expression of IL-1β, IL-6 and TNF- α in the gastric tissue when same was compared to normal control group (NC). Pretreatment of DC group with different doses of vincamine (VLD, VMD and VHD) significantly downregulated (all P < 0.05) the mRNA expressions of IL-1β, IL-6 and TNF- α and ameliorated oxidative stress marker MDA and increased antioxidant markers like SOD and GSH in the gastric tissue when same was compared to the DC group. In a nutshell, vincamine provide gastric protection in the BALB/c mice of gastric ulcer group by increasing the gastric pH, attenuated total acidity of the stomach and modulated infiltration of inflammatory cells, edema, fibrinoid necrosis, hemorrhage, and erosion score when compared to the DC group. Furthermore, vincamine possesses antiulcer and gastroprotective activity which may be ascribed to down-regulation the mRNA expression of IL-1β, IL-6 and TNF- α in the gastric tissue of disease control group. Vincamine also provide gastroprotective role by increasing the concentration of SOD and GSH while decreasing the MDA in gastric tissue.

To study sufentanil’s effect on the proliferation, apoptosis, and epithelial-mesenchymal transition (EMT) of ovarian cancer cells by modulating the SMAD3/SNAIL signaling pathway. Ovarian cancer A2780 cells were exposed to sufentanil at varying concentrations of 0 ng/mL, 20 ng/mL, 40 ng/mL, 60 ng/mL, 80 ng/mL, and 100 ng/mL. The proliferation of A2780 cells was assessed using the CCK-8 method; A2780 cells were randomly divided into three groups: CON group (normal culture), SUF group (60 ng/mL sufentanil intervention), and SUF + ACA group (60 ng/mL sufentanil and 100 ng/mL activin A intervention). After 48 h of culture, cell migration was evaluated by the scratch assay; Cell invasion was assessed using the Transwell chamber assay; Cell apoptosis was measured via flow cytometry; The growth status of the cells was observed under an optical microscope; The expression of N-cadherin, E-cadherin, SMAD3, TGF-β, and SNAIL proteins was detected by Western blot. When the concentration of sufentanil was ≥ 20 ng/mL, it dose-dependently inhibited the proliferation of ovarian cancer A2780 cells; In comparison to the CON group, the number of A2780 cells in the SUF group was significantly reduced, some cells detached, cell migration and invasion abilities, and the expression levels of N-cadherin, SMAD3, TGF-β, and SNAIL proteins decreased, while the apoptosis rate and E-cadherin protein expression levels increased (P < 0.05); In comparison to the SUF group, the growth status of A2780 cells in the SUF + ACA group was good, cell migration and invasion abilities, and the expression levels of N-cadherin, SMAD3, TGF-β, and SNAIL proteins increased, while the apoptosis rate and E-cadherin protein expression levels decreased (P < 0.05). Sufentanil may inhibit the proliferation and epithelial-mesenchymal transition of ovarian cancer cells and promote cell apoptosis by inhibiting the SMAD3/SNAIL signaling pathway.

SARS-CoV-2 E and 3a proteins are important for the assembly, budding, and release of viral particles. These two transmembrane proteins have been implicated in forming channels in the membrane that allow the transport of ions to favor viral replication. During an active infection, both proteins generally localize to the endoplasmic reticulum (ER), ER-Golgi intermediate compartment (ERGIC), and the Golgi where viral assembly occurs. The ER and Golgi are critical for the proper packaging and trafficking of cellular proteins along the secretory pathways which determine a protein’s final destination inside or outside of the cell. The SARS-CoV-2 virus primarily infects epithelial cells that are highly secretory in nature such as those in the lung and gut. Here we quantified the distribution of SARS-CoV-2 E and 3a proteins along the secretory pathways in a human intestinal epithelial cell line. We used NaturePatternMatch to demonstrate that epitope-tagged E and 3a proteins expressed alone via transient transfection have a similar immunoreactivity pattern as E and 3a proteins expressed by wild-type viral infection. While E and 3a proteins localized with all selected cellular markers to varying degrees, 3a protein displayed a higher correlation coefficient with the Golgi, early/late endosome, lysosome, and plasma membrane when compared to E protein. This work is the first to provide quantification of the subcellular distribution of E and 3a proteins along the multiple components of the secretory pathway and serves as a basis to develop models for examining how E and 3a alter proteostasis within these structures and affect their function.

Nebivolol (NB), which is a commonly used β₁ adrenoreceptor blocker, shows protective effects against oxidative stress and inflammation-related processes. In this study, we aimed to evaluate the possible protective effects of NB via the vascular cell adhesion molecule-1/matrix metalloproteinases-9 (VCAM-1/MMP-9) signaling pathway on systemic inflammation induced testicular dysfunction. Four groups of 32 male Wistar Albino rats were divided as (n = 8 for each) the control; lipopolysaccharide (LPS; 5 mg/kg on the third day); LPS + NB (NB: 10 mg/kg for three days and 5 mg/kg LPS 30 min following the last NB dose); NB (10 mg/kg for three days). Six hours following the LPS administration, rats were sacrificed, then testicular tissues were collected for evaluating total oxidant status (TOS), total antioxidant status and oxidative stress index (OSI) levels biochemically, VCAM-1 and MMP-9 mRNA expression levels, caspase-3 (cas-3), tumor necrosis factor-alpha (TNF-α) expressions by immunohistochemically. Systemic inflammation caused significant increases in TOS and OSI levels, VCAM-1, MMP-9, cas-3, TNF-α expressions, and a decrease of the spermatozoa count compared to the control group. NB administration successfully restored all these changes significantly. Thus, NB can be a protective drug candidate for testicular dysfunction secondary to systemic inflammation with its potent antioxidant and anti-inflammatory mechanisms.

Dental pulp stem cells (DPSCs), a subset of tooth-derived mesenchymal stem cells (MSCs), demonstrate significant promise in clinical stem cell therapy. However, prolonged in vitro expansion commonly results in compromised stemness, limiting therapeutic efficacy. Thus, maintaining the stemness of DPSCs during expansion and culture is a key challenge for regenerative medicine. In the current study, the impact of simulated microgravity (SMG) on DPSC stemness was investigated using the three-dimensional clinostat Cellspace-3D. After SMG treatment for 3 days, DPSCs demonstrated markedly enhanced replicative activity, proliferation efficiency, self-renewal capacity, and effective inhibition of the senescence process. Under specific differentiation induction conditions, DPSCs in the SMG group exhibited superior osteogenic, adipogenic, chondrogenic, and neural differentiation potentials. Additionally, DPSCs exhibited higher expression levels of the MSC surface markers Stro-1 and CD146 and stemness maintenance-related genes Oct4, Nanog, and Sox2 in the SMG group compared to those from the normal gravity (NG) group. To elucidate the potential molecular mechanisms by which SMG influences the stemness of DPSCs, transcriptome sequencing of total RNA was performed, and identified that differentially expressed genes (DEGs) are closely associated with the MAPK signaling pathway. Further verification experiments demonstrated that the MAPK/ERK signaling pathway was activated in the SMG group. In conclusion, SMG effectively maintains the stemness of DPSCs cultivated in vitro, and its mechanism of action may be associated with the activation of the MAPK/ERK signaling pathway.

Non-small cell lung cancer (NSCLC) is a life-threatening disease that still lacks effective targeted treatment. Family with sequence similarity 64, member A (FAM64A) is dysregulated in different malignancies and participates in the cancer progression. To address the role of FAM64A in NSCLC. The FAM64A expression was detected by reverse transcription quantitative polymerase chain reaction and western blotting. Short-hairpin RNAs (shRNAs) against FAM64A were transfected into A549 and H460 cells. The role of FAM64A in vitro was evaluated by cell counting kit-8, transwell, enzyme-linked immunosorbent assay, immunofluorescence and western blotting. In vivo role of FAM64A was addressed in xenografted mice using immunohistochemistry and western blotting. FAM64A was upregulated in NSCLC that predicted a dismal prognosis for NSCLC patients. Knockdown of FAM64A diminished cell viability, invaded cell numbers, the Vimentin expression and the concentrations of TGF-β and IL-10, but fostered the E-cadherin expression and the concentrations of IL-2 and IFN-γ in NSCLC cells. Mechanistically, silencing of FAM64A declined the expression of JAK/STAT3/PD-L1 pathway, which was restored with the application of RO8191, an activator of JAK/STAT3 pathway. The inhibitory role of FAM64A knockdown in NSCLC cell proliferation, invasion, EMT and immune escape was inverted by the management of RO8191. In vivo, knockdown of FAM64A reduced tumor size and weight, the level of Vimentin and PD-L1, and the expression of JAK/STAT3/PD-L1 pathway, but enhanced the IFN-γ level. Upregulation of FAM64A promoted proliferation, invasion, EMT and immune escape through activating JAK/STAT3/PD-L1 axis.