Cytotechnology最新文献

Acute infective endocarditis (AIE) is an inflammatory reaction caused by the infection of the endocardial surface, accompanied by the formation of vegetation with different shapes and sizes. miR-361-5p and secretory leukocyte protease inhibitor (SLPI) were reported to mediate the process of staphylococcus aureus (SA) infection in AIE, but their roles in AIE are unclear. In this study, the AIE cell model was induced by hypoxia and infection of SA, and the AIE rat model was established. Dual-luciferase experiment was performed to verify the interaction between miR-361-5p and SLPI. Reverse transcription real-time polymerase chain reaction (RT-qPCR), western blot, Cell counting kit-8, flow cytometry, and Enzyme-linked immunosorbent assay (ELISA) kits were used to detect the role of miR-361-5p and SLPI in AIE cells. Hematoxylin and eosin staining, RT-qPCR, western blot, Terminal deoxynucleotidyl transferase dUTP nick end-labeling staining and ELISA kits were used to detect the function of miR-361-5p and SLPI in AIE rats. Mechanistic research revealed that miR-361-5p targeted SLPI. miR-361-5p was highly expressed in AC16 cells induced by hypoxia and SA, while SLPI expression was decreased. Compared with the NC group, hypoxia and SA infection-induced AIE model cells exhibited significantly reduced cell viability, elevated apoptosis rates, increased concentrations of creatine kinase-MB (CK-MB), cardiac troponin T (cTnT), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α), as well as activation of nuclear factor-kappa B p65 (NF-κB p65). Knockdown of miR-361-5p or overexpression of SLPI increased AC16 cell viability, reduced AIE-induced apoptosis rates, decreased CK-MB, cTnT, IL-1β, and TNF-α levels, and suppressed NF-κB p65 activation. Knockdown of SLPI reversed the effects of miR-361-5p knockdown on AC16 cells. In vivo results demonstrated that knockdown of miR-361-5p suppressed valvular vegetation formation in rats with AIE, reduced myocardial apoptosis, and decreased levels of CK-MB, cTnT, IL-1β, and TNF-α. Our study demonstrates that knockdown of miR-361-5p has a protective effect on AIE by promoting SLPI and inhibiting NF-κB P65 signalling pathway, which may provide a new perspective for treating AIE.

Different extract products of Capparis ovata species have antimicrobial, antidiabetic, antineuroinflammatory, and also hepatotoxicity and oxidative stress inhibitory effects. In our study, we aimed to compare the cytotoxic effects of polysaccharides isolated from Capparis ovata, on two different colon cancer cell lines by VEGF and GSK-3β signaling. Caco-2 and HT-29 colon cancer cells were treated with Capparis ovata polysaccharides and doxorubicin at varied concentrations for 24 and 48 h. The proliferation levels were measured by MTT assay, expression of VEGF and GSK-3β molecules determined using qRT-PCR assay and immunocytochemical staining. And the scratch wound model was used the determined the effect of polysaccharides on cell migration. The data was analyzed statistically. It was found that cell proliferation and migration were suppressed by Capparis ovata polysaccharides concentration dependent manner. Also Capparis ovata polysaccharides significantly decreased the expression of VEGF and GSK-3β molecules in both cells compared with the untreated cells. According to our findings, Capparis ovata polysaccharides showed cytotoxic effect on Caco-2 and HT-29 cells concentration dependent manner, by inhibiting the expression of VEGF and GSK-3β which play a role in tumor formation and metastasis. So, cell migration was repressed.

Secreted phosphoprotein 1 (SPP1) has been identified as a driver of fibrosis and inflammation, while its contribution to calcific aortic valve disease (CAVD) has not been clarified. In this research, we explored the possible role of SPP1 and the underlying molecular mechanism in CAVD. C57BL/6J mice were subjected to a high-fat diet (HFD), and human valve interstitial cells (VICs) were induced with the osteogenic medium (OM) for modeling. SPP1 was highly expressed in the human calcific aortic valve and VICs induced with OM, and its transcription was regulated by EP300/CREBBP. Knockdown of SPP1 inhibited the osteogenic differentiation of VICs, and overexpression of SPP1 reversed the repressive effect of EP300/CREBBP inhibitor on the osteogenic differentiation of VICs. EP300/CREBBP inhibitor ameliorated the aortic valve thickening and calcification in mice induced with HFD, and combined overexpression of SPP1 reversed the effect. In conclusion, the present findings suggest that EP300/CREBBP is dependent on SPP1 to induce osteogenic differentiation of VICs, thereby promoting the development of CAVD.

The tight junctions (TJs) between nasal mucosal epithelial cells are a crucial component of the nasal barrier function. Incomplete formation or reduced expression of TJs is a primary contributor to the onset and progression of allergic rhinitis (AR). Therefore, an in-depth investigation into the mechanisms affecting the barrier function of human nasal mucosal epithelial cells (HNEpCs) may facilitate the identification of new therapeutic approaches for AR treatment. Bioinformatics analysis found RGS1 is upregulated in AR, but its impact on the nasal mucosal epithelial barrier function remains unclear. This study aims to explore the mechanism of RGS1 regulating epithelial barrier function in AR. Differentially expressed genes in AR were analyzed using GSE43523 from GEO database. RGS1 expression level was validated in AR clinical samples and IL-13-induced HNEpCs. Loss and function of RGS1 or/and AQP5 was performed in IL-13-induced HNEpCs to detect the activation of NF-κB signal pathway. The epithelial barrier function of HNEpCs was measured by trans-epithelial electrical resistance (TER) and FITC-Dextran 4(FD4) assay. TJs, such as ZO-1, Occludin and Claudin-1 were also detected by western blot and Immunofluorescence. Bioinformatics analysis, AR clinical samples and IL-13-induced HNEpCs consistently found up-regulated RGS1 expression in AR. RGS1 silencing can protect HNEpCs against IL-13-induced epithelial barrier dysfunction, evidence by increased TER value, decreased FD4 and elevated expression of ZO-1, Occludin and Claudin-1. RGS1 silencing can also suppress the activation of NF-κB signal pathway and increase AQP5 expression, which such expression pattern can be nullified in response to AQP5 silencing. RGS1 was found to be elevated in AR. Silencing of RGS1 can suppress NF-κB signal pathway to increase AQP5 expression, thereby attenuating epithelial barrier dysfunction in HNEpCs.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00825-4.

Oxymatrine is a quinolizidine alkaloid derived from Sophora roots that has demonstrated significant antitumor activity against various cancers, including lung cancer. Recently, combination therapies involving anticancer agents and targeted interventions for dysregulated genes have emerged as a promising strategy to enhance treatment efficacy and overcome drug resistance. This study investigates the synergistic effects of oxymatrine and GIMAP8 in modulating the progression of lung adenocarcinoma (LUAD). Through bioinformatics analysis and qRT-PCR, GIMAP8 was identified as a key downregulated gene in LUAD, with reduced expression confirmed in LUAD samples and cell lines. Functional cell experiments showed that GIMAP8 overexpression effectively suppressed the malignant characteristics of LUAD cells by regulating the Wnt/β-catenin pathway and inducing cell cycle arrest. Similarly, oxymatrine treatment significantly inhibited these malignant properties, and an even stronger synergistic effect was observed when oxymatrine treatment was combined with GIMAP8 overexpression, both in vitro and in vivo. In conclusion, these findings demonstrate that the combination of oxymatrine and GIMAP8 exerts synergistic antitumor effects in LUAD cells by modulating the Wnt/β-catenin pathway, supporting its potential as an effective therapeutic strategy for LUAD.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00839-y.

Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive interstitial pneumonia of unknown etiology, characterized by recurrent acute lung injury. This leads to worsening dyspnea and lung function decline. Patients with idiopathic pulmonary fibrosis (IPF) generally have a poor prognosis, often succumbing within 2-3 years of diagnosis, and the five-year survival rate is below 40%. Analysis of gene differences between raw and normal samples in the database, along with gene ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, suggests that ferroptosis may play a role in the development of pulmonary fibrosis. To test this hypothesis, transforming growth factor beta 1 (TGF-β1) was used to induce fibrosis in vitro, and levels of pulmonary fibrosis markers such as alpha-smooth muscle actin (α-SMA), fibronectin (FSN), and collagen (COL) were measured by enzyme-linked immunosorbent assay (ELISA). The results confirmed successful establishment of the pulmonary fibrosis model in vitro and indicated increased levels of ferroptosis markers, including glutathione (GSH) and iron accumulation. Salidroside, an extract from Rhodiola rosea, has been found to enhance glutathione levels in lung tissue, reduce oxidative stress, and decrease levels of α-SMA, FSN, COL, GSH, and iron in the model. These findings suggest that salidroside can mitigate pulmonary fibrosis by reversing ferroptosis. Additionally, we observed high expression of phospholipase A2 group IVA (PLA2G4A) in the fibrosis model, which was reduced by salidroside, indicating that salidroside may regulate ferroptosis via the PLA2G4A gene.

To investigate the function and mechanism of action of LncRNA NR2F1-AS1 involved in osteogenic differentiation process. An in vitro model was constructed by osteogenic differentiation-induced stimulation (OS) on the hFOB1.19 cell line. Real-time fluorescence quantitative PCR (RT-qPCR) was performed to detect the expression of NR2F1-AS1, miR-423-5p and osteogenic differentiation markers (RUNX2, OCN, OPN). Cell Counting Kit-8 (CCK8) method and flow cytometry were observed cell proliferation and apoptosis, respectively. Enzyme linked immunosorbent assay (ELISA) tested alkaline phosphatase (ALP) activity. Dual-Luciferase Report (DLR) assay and RNA immunoprecipitation (RIP) verified gene interactions. Bioinformatics methods predicted downstream target genes and their pathways of action. OS increased osteogenic differentiation markers and NR2F1-AS1 expression and decreased miR-423-5p levels. Transfection of si- NR2F1-AS1 promoted OS osteoblast apoptosis, but inhibited cell proliferation, ALP activity and osteogenic differentiation marker expression. NR2F1-AS1 is mostly present in the cytoplasm and is involved in the osteogenic differentiation process by down-regulating miR-423-5p. The use of miR-423-5p inhibitor can resist apoptosis induced by silencing NR2F1-AS1, promote osteoblast proliferation, activate ALP activity, and induce osteogenic differentiation process in osteoblasts. Bioinformatics prediction identified 82 target genes that might be involved in osteogenic differentiation, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis suggested that they were mainly associated with inter-synaptic formation and signaling. NR2F1-AS1 may promote osteoblast proliferation stimulate ALP activity, and induce osteogenic differentiation by down-regulating miR-423-5p.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00786-8.

Preeclampsia (PE) is one of the most common and serious documented gestational complications, and it is threatening the mother and the fetus, which is a notable burden on healthcare systems. Aldehyde dehydrogenase 1A1 (ALDH1A1), a cytosolic enzyme, shows vital physiological and pathophysiological functions in many areas. In the majority of cancer types, obesity-associated protein (FTO) is upregulated and exhibits an essential tumor-promoting role. We speculate that FTO and ALDH1A1 may play a significant role in the pathogenesis of PE by affecting the trophoblast cell biological behaviors. We analyzed differential expression genes (DEGs) in PE and non-PE groups in the GSE234726 dataset. The reverse-transcription quantitative polymerase chain reaction (qRT-PCR) and western blot assay were performed to test the mRNA and protein levels. The cell proliferation and apoptosis were examined using 5-Ethynyl-2'-deoxyuridine (EdU) and flow cytometry. The cell migration was investigated by wound healing assay and transwell assay. The ability of angiogenesis was tested by angiogenesis assay. The Spearman's rank correlation coefficient was used to analyze the correlation between ALDH1A1 expression and FTO expression. The m6A methylation site of ALDH1A1 mRNA was predicted using SRAMP website. The RNA immunoprecipitation (RIP) and m6A RNA immunoprecipitation (MeRIP) assay were performed to examine the binding relationship between ALDH1A1 and FTO. In PE, ALDH1A1 level is decreased. Silencing ALDH1A1 suppressed cell proliferation, migration, and angiogenesis and induced cell apoptosis. ALDH1A1 knockdown inhibited the expression of cyclin D1, anti-matrix metalloproteinase 9 (MMP9), and vascular endothelial growth factor (VEGF) and facilitated c-casp3 levels. The FTO expression was increased in PE placentas. Besides, the ALDH1A1 expression was negatively correlated with FTO levels, and FTO could target ALDH1A1. Mechanically, FTO repressed the biological behaviors of HTR-8/SVneo cells via ALDH1A1 down-regulation. FTO retards the HTR-8/SVneo cell biological function through knockdown of ALDH1A1. These results suggest that FTO and ALDH1A1 may play an important role in the pathogenesis of PE.

Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are respiratory conditions associated with high mortality rates, primarily due to intense pulmonary inflammation. Lipopolysaccharide (LPS), an essential constituent of the wall of gram-negative bacteria, has the potential to trigger inflammation, ultimately leading to ALI. This study assessed the protective role of selegiline in mitigating LPS-induced inflammation in A549 cells. A549 cells were divided into seven groups: the negative control (PBS, 100 μM), the positive control (H₂O₂, 100 μM), the LPS group (1 µg/ml), groups treated with selegiline (Sel) (20, 30, and 40 μM) + LPS, and a group receiving Sel (40 μM). Parameters, including cell viability, iNOS activity, levels of SOD, CAT, and GPx, along with the pro-inflammatory cytokines IL-1β and TNF-α, as well as the protein expressions of NF-kB, NLRP3, and Caspase-1, were analyzed. Findings revealed that exposure to LPS increased oxidation and inflammation in A549 cells. Conversely, Sel markedly enhanced antioxidant capacity and diminished the iNOS activity, IL-1β, and TNF-α levels, as well as reduced the expression of inflammatory markers, including NF-kB, NLRP3, and Caspase-1 proteins. These results indicate that Sel may ameliorate LPS-induced pulmonary inflammation through its beneficial properties, including antioxidant and anti-inflammatory effects.

Sevoflurane, a widely utilized inhalational anesthetic, has been shown in previous studies to inhibit the proliferation and invasion of non-small cell lung cancer (NSCLC) cells. However, whether sevoflurane affects the metastasis of small cell lung cancer (SCLC) cells remains unexplored. This study's primary objective was to investigate the mechanism by which sevoflurane induces calcium overload and EFHD1 upregulation, thereby contributing to pyroptosis in SCLC cells. NCI-H446 cells were treated with sevoflurane at concentrations of 0%, 0.6%, 3% and 15%. Cell proliferation was assessed using the CCK8 assay, while cell migration and invasion were analyzed using the Transwell assay. EFHD1 expression was determined by q-PCR and Western blot. Calcium content, ROS levels, and CCO content were measured using respective assay kits, and mitochondrial morphology was observed using transmission electron microscopy. Western blotting was performed to detect the expression of caspase-1 and GSDMD. All procedures were repeated under conditions of EFHD1 overexpression or silencing. Sevoflurane inhibited the proliferation, migration, and invasion of NCI-H446 cells, concurrently exerting a significant pro-pyroptotic effect. This was mediated by mitochondrial calcium overload, EFHD1 upregulation, and subsequent mitochondrial damage-induced pyroptosis. Sevoflurane significantly enhances pyroptosis in NCI-H446 cells, thereby potentially reducing cancer cell dissemination during surgery.