Cytotechnology最新文献

Glycation of low-density lipoprotein (LDL) occurs when blood glucose levels are high, as in diabetic patients. This can lead to abnormal cholesterol transport in the body. Glycated LDL (G-LDL) is harmful and triggers oxidative stress in human erythrocytes. The present study investigated the protective effect of crocin, found in saffron, against G-LDL-induced cytotoxicity, oxidative damage, and formation of reactive species in human erythrocytes. These parameters were assessed in isolated human erythrocytes exposed to 6 mg/ml G-LDL, with and without varying concentrations of crocin (0.5, 1.0, 1.5 mM). Increased hemolysis, methemoglobin, and oxoferrylHb were seen in G-LDL alone-incubated cells. A significant increase in reactive species in G-LDL-exposed erythrocytes led to enhanced oxidation of lipids, proteins, and thiols. The activities of certain key antioxidant and membrane-bound enzymes were reduced. The antioxidant capacity of cells was compromised as indicated by a diminished ability to neutralize free radicals and donate electrons. G-LDL significantly altered surface morphology, forming echinocytes and agglutinating the cells. All these characteristics were significantly restored when erythrocytes were pre-treated with crocin, before incubation with G-LDL, in a crocin concentration-dependent manner. Furthermore, erythrocytes incubated with 1.5 mM crocin alone did not show alterations in any of the above parameters, indicating that crocin was not toxic to these cells. These results clearly show that crocin is strongly cytoprotective against G-LDL-induced damage and toxicity in erythrocytes. Hence, it can be used as an efficient dietary antioxidant in various atherosclerotic cardiovascular disorders, as seen in diabetic patients.

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Sepsis, a life-threatening systemic inflammatory response syndrome mediated by dysregulated host-pathogen interactions, frequently precipitates in renal injury. Renal tubular epithelial cell (RTEC) injury is a hallmark of septic acute kidney injury (AKI). Recent studies have demonstrated the involvement of serum response factor (SRF) in septic AKI. Herein, the role and mechanism of SRF as a transcription factor in regulating RTEC dysfunction were explored. HK-2 cells (the human RTEC line) were treated with lipopolysaccharide (LPS) for the establishment of in vitro septic AKI models. HK-2 cell viability was validated using CCK-8 assay. HK-2 cell apoptosis was evaluated by flow cytometry analysis. Measurement of proinflammatory cytokine concentration was conducted using enzyme-linked immunosorbent assay kits. RT-qPCR were required for determining gene levels. Western blotting was prepared for testing the protein levels of proinflammatory cytokines, apoptosis-related markers, NF-κB p65 and NF-κB inducing kinase (NIK). The binding of SRF to NIK promoter was confirmed by chromatin immunoprecipitation and luciferase reporter assays. LPS treatment suppressed HK-2 cell viability and accelerated HK-2 cell inflammation and apoptosis, which was antagonized by SRF depletion. Mechanically, SRF as a transcription factor bound to NIK promoter. SRF silencing inhibited LPS-induced NF-κB signaling activation in HK-2 cells. In rescue assays, NIK overexpression counteracted the restrictive impact of SRF deficiency on LPS-induced HK-2 cell dysfunction. SRF aggravates LPS-elicited HK-2 cell injury via binding to NIK promoter and activating NF-κB signaling.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00854-z.

Diabetic kidney disease (DKD) is a leading cause of kidney failure. It is characterized by damage to renal tubular endothelial cells, which is exacerbated by macrophage-derived exosomes under high-glucose (HG) conditions. Triptolide (TPL), an active ingredient extracted from Tripterygium wilfordii Hook F, has been used in the treatment of DKD. In this study, we aimed to investigate whether TPL protects against renal damage induced by macrophage-derived exosomes under HG conditions and to explore the involvement of the protein kinase B/mammalian target of rapamycin (AKT/mTOR) signaling pathway in this process. Exosomes were isolated from macrophages cultured in normal glucose (NG) and HG environments and characterized using transmission electron microscopy, nanoparticle tracking analysis, and Western blot. Subsequent in vitro experiments on mouse renal tubular epithelial cells (mRTECs) demonstrated that 5 ng/mL TPL enhanced cell viability, reduced apoptosis and inflammation, and promoted autophagy-effects that were reversed by the autophagy inhibitor chloroquine (CQ). Molecular docking and bioinformatic analyses suggested a stable interaction between TPL and AKT, implicating the AKT/mTOR pathway in TPL-mediated autophagy activation. Further in vivo studies using an AKT agonist supported the role of TPL in inhibiting AKT/mTOR signaling and enhancing autophagic activity, ultimately ameliorating renal injury in a DKD model. These findings provide novel insights into the mechanisms by TPL may mitigate exosome-induced renal damage, highlighting its potential as a therapeutic agent for DKD.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00883-8.

Microglial polarization plays a crucial role in Parkinson's disease (PD). This study explores how serpin family A member 1 (SERPINA1) suppresses neuroinflammation and alleviates neuronal damage in PD. Adeno-associated viruses were injected into mice to manipulate the expression of SERPINA1 or runt-related transcription factor 1 (RUNX1) in the substantia nigra, followed by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) modeling. Behavioral tests, histopathology (HE and Nissl staining), immunohistochemistry (IHC), immunofluorescence, and enzyme-linked immunosorbent assay were conducted to evaluate neuroinflammation and neuronal damage in mice. BV2 microglial cells were infected with lentiviruses overexpressing SERPINA1 and treated with 100 µM 1-methyl-4-phenyl-pyridinium (MPP)⁺. MPP⁺ increased pro-inflammatory cytokines and iNOS while decreasing anti-inflammatory cytokines and arginase-1 expression in BV2 cells. SERPINA1 and RUNX1 were upregulated in the SN of MPTP-induced mice. RUNX1 bound to the promoter region of SERPINA1 to induce its transcription. SERPINA1 or RUNX1 overexpression alleviated PD-related neuronal damage and neuroinflammation in mice and MPP⁺-induced inflammation in BV2 cells. SERPINA1 knockdown inhibited M2 polarization in the presence of RUNX1 overexpression. Taken together, RUNX1 transcriptionally activates SERPINA1, promoting microglial M2 polarization, suppressing neuroinflammation, and alleviating neuronal damage in PD.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00878-5.

Oral squamous cell carcinoma (OSCC) is an aggressive malignancy with poor prognosis and limited therapeutic options. Plasminogen activator, urokinase (PLAU) is emerged as a potential key player in OSCC by bioinformatics analysis, but its function in OSCC have not been explored. Therefore, this study investigates its role and mechanism in OSCC progression, with a focus on its interaction with the N6-methyladenosine (m⁶A) writer ZC3H13. Differentially expressed genes in OSCC were identified through analysis of GEO datasets. A candidate oncogene was subsequently selected based on Gene Ontology enrichment analysis and validated using qRT-PCR in patient samples. The role of PLAU was examined by CCK-8, EdU, and transwell assays. In vivo tumorigenicity was evaluated using xenograft models. The relationship amongst PLAU and ZC3H13 was analyzed through correlation analysis, MeRIP, RIP, qRT-PCR, immunoblotting and mRNA stability assays. Finally, rescue experiments were conducted to validate the interactions between ZC3H13 and PLAU. PLAU upregulated in OSCC was identified as a candidate oncogene based on bioinformatic analysis and mRNA expression profiling. PLAU silencing suppressed cell proliferation, migration, invasion, and tumor growth, whereas its overexpression produced the opposite effects. Furthermore, ZC3H13 was positively correlated with PLAU expression. ZC3H13 overexpression enhanced m⁶A levels of PLAU to increase PLAU expression and mRNA stability. In addition, ZC3H13 overexpression partially rescued the suppressive effects of PLAU silencing on OSCC cells. In summary, these results illustrate that ZC3H13-mediated m⁶A modification increases PLAU mRNA stability and expression, thereby enhancing OSCC progression.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00882-9.

Gastric cancer (GC) is one of the most common malignant tumors worldwide. Banxia Xiexin Decoction (BXXXD) has a significant therapeutic effect on digestive system diseases, but its mechanism of action on GC is not yet clear. Using single-cell and bioinformatics to discover prognostic markers and Tumor Microenvironment (TME) components for GC, and further explore the molecular mechanism of BXXXD in treating GC through network pharmacology. KIF2C, KIF20A, KIF11, CDK1, CDC20, FN1, etc. could be used as diagnostic and prognostic markers for GC. The tumor microenvironment of GC mainly includes T cell, Neutrophil, B cell, Macrophage, Epithelial cell, Plasma cell, Stromal cell, Fibroblast, common myeloid progenitor cell. The core active ingredients of BXXXD synergistically regulate the MAPK signaling pathway, oxidative stress response, and apoptosis pathway, and multi-target reverse the pathological process of GC. We have established the correlation between the pathogenesis of GC's "cold and heat disorder" and modern pathological indicators (inflammation, oxidative damage, and cell apoptosis disorder) from the perspective of traditional Chinese medicine syndrome, providing a theoretical basis for the clinical application of BXXXD.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00888-3.

The anti-proliferation, anti-invasion, and anti-migration activities of KLF2 in keloid fibroblasts (KFs) and hypertrophic scar fibroblasts (HSFs) were investigated. KLF2 in normal skin (NF), keloid (KD), hypertrophic scar (HS) tissues, and cell lines was quantified using western blot and quantitative real-time polymerase chain reaction techniques. The processes of cell proliferation, migration, and invasion were observed in both KFs and HSFs, with pathway proteins linked to these processes pinpointed through western blot analysis. KLF2 in KD and HS tissues was lower than that in NF tissues. Enhancing KLF2 expression inhibited KF and HSF proliferation, migration, and invasion. MMP2, MMP-9, PI3K and p-Akt protein levels were inhibited in KFs with KLF2 overexpression. However, inhibition of PI3K and p-Akt protein levels was observed only in KLF2-overexpressed HSFs. In KFs with enhanced KLF2 expression, PI3K agonists eliminated the effect of KLF2 overexpression on cell migration and invasion. KLF2 inhibits the proliferation and migration of KFs by down-regulating MMP-2/9 through the PI3K/AKT pathway, suggesting that KLF2 may be a potential therapeutic target for KD. Hence, these findings offer novel perspectives on the function and molecular pathways of KLF2 in KD, as well as novel strategies for its clinical treatment.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00885-6.

Atherosclerosis (AS) is linked to many cardiovascular disorders. We investigated the potential roles and mechanisms of microRNA (miR)-218-5p in AS. Primary mouse aortic endothelial cells (MAECs) were induced with ox-LDL, followed by various interventions including miR/gene overexpression and knockdown. An AS mouse model was established in ApoE^-/- mice and treated with miR-218-5p agomir. Serum lipid, inflammatory factor, interleukin (IL)-1β, IL-18, and pyroptosis-related protein levels as well as plasma anti-inflammatory IL-10 and TGF-β levels were determined. The results demonstrated that miR-218-5p overexpression decreased tumor necrosis factor receptor superfamily member 11 A (TNFRSF11A) protein level and ameliorated ox-LDL-induced MAEC pyroptosis. Furthermore, miR-218-5p impeded the nuclear factor (NF)-κB signaling and inactivated NOD-like receptor protein 3 (NLRP3)/caspase-1 via TNFRSF11A. NLRP3 activation partially reversed the impacts of miR-218-5p on pyroptosis. We observed augmented levels of serum total cholesterol, triglyceride, low-density lipoprotein cholesterol, and pro-inflammatory proteins, elevated levels of N-terminus of gasdermin D, TNFRSF11A, nuclear NF-kB p65, phosphorylated p65, NLRP3, and cleaved-caspase-1 proteins in aortic tissue, and reduced serum high-density lipoprotein cholesterol level and cytoplasmic NF-kB p65 protein level in AS-rendered mice. miR-218-5p agomir treatment reduced cell pyroptosis in aorta of AS model mice and improved AS. Briefly, miR-218-5p repressed TNFRSF11A, repressed the NF-κB signaling, and disrupted NLRP3/caspase-1 activation, thereby alleviating pyroptosis and contributing to the improvement of AS.

Preeclampsia (PE) is a pregnancy-specific disorder characterized by new-onset hypertension after 20 weeks of gestation, and is associated with abnormal placental development, placental ischemia, and systemic endothelial dysfunction. There is new evidence that ferroptosis, a kind of cell death that depends on iron, plays a role in placental injury in PE. Unfortunately, our understanding of the regulatory mechanisms that underlie ferroptosis in PE is still limited. The purpose of this research was to examine miR-150-3p's function as a diagnostic biomarker and its function in controlling ferroptosis in the placenta of patients with PE via modulating ferritin heavy chain 1 (FTH1). We compared clinical indicators and placental ferroptosis markers in PE and normal pregnant women. Bioinformatics tools were used to predict miRNAs targeting ferroptosis-related genes, and experimental validation was performed in placental tissues and trophoblast cells. Dual-luciferase reporter assays and cell function analyses were conducted to examine the role of miR-150-3p and its target FTH1. PE placentas exhibited increased iron content and reduced expression of GPX4, SLC7A11, and FTH1. Bioinformatics identified miR-150-3p, miR-27a-3p, and miR-214-3p as top regulators of FTH1, SLC7A11, and GPX4, respectively. MiR-150-3p was markedly upregulated in PE and showed high diagnostic accuracy (AUC = 0.868). Functional assays revealed that miR-150-3p promotes ferroptosis in trophoblast cells by directly downregulating FTH1, leading to increased iron accumulation, lipid peroxidation, and oxidative stress. Our findings reveal a novel molecular pathway involving miR-150-3p/FTH1 in the development of PE, offering insights into potential biomarkers and therapeutic strategies targeting ferroptosis.

Endometrial cancer (EC) is one of the most common gynecological malignancies, and its progression is tightly linked to extracellular matrix (ECM) remodeling and metabolic reprogramming. Collagen type XXIII alpha 1 (COL23A1), a transmembrane collagen, has been implicated in several cancers, but its expression pattern, functional role and upstream regulation in EC remain unclear. Public datasets (TCGA, GTEx, cBioPortal) were analyzed to characterize COL23A1 expression, clinicopathological correlations and prognostic value. The biological functions of COL23A1 in EC cells were assessed by qRT-PCR, Western blotting, CCK-8, colony formation, flow cytometry, Transwell assays and Seahorse extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) measurements. Untargeted metabolomics and RNA immunoprecipitation (RIP) were used to interrogate glycolytic metabolism and m⁶A modification. Xenograft models were established to validate the in vivo effects of COL23A1. COL23A1 was significantly upregulated in EC tissues and correlated with advanced clinicopathological features and poor overall survival. Genetic silencing of COL23A1 suppressed EC cell proliferation, clonogenicity, migration and invasion in vitro, and inhibited tumor growth in vivo. At the metabolic level, COL23A1 knockdown disrupted glycolytic metabolism, leading to reduced glucose uptake, lactate production, expression of key glycolytic enzymes and ECAR, accompanied by a compensatory increase in OCR. Integrative bioinformatic and experimental analyses showed that METTL14 installs m⁶A modifications on COL23A1 mRNA, whereas the m⁶A reader YTHDF1 binds and stabilizes the modified transcript, thereby sustaining COL23A1 expression. Rescue experiments demonstrated that COL23A1 is required for METTL14- and YTHDF1-driven glycolytic reprogramming and oncogenic phenotypes in EC cells. COL23A1 acts as a previously unrecognized oncogenic driver in endometrial cancer, promoting tumor progression and glycolysis-dependent metabolic reprogramming through a METTL14-m⁶A-YTHDF1-COL23A1 axis. Targeting this m⁶A-dependent pathway may offer a promising therapeutic strategy for endometrial cancer.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-026-00898-9.