Experimental cell research最新文献

Oocyte quality plays a fundamental role in fertilization and embryonic development. Emerging evidence indicates that ferroptosis may impair oocyte quality. Ferroptosis suppressor protein 1 (FSP1), a known ferroptosis inhibitor, has an uncharacterized function in regulating oocyte quality regulation during meiotic maturation. This study identified FSP1 expression across all stages of meiotic maturation with localization to the cytoplasm of mouse oocytes. Aged mice exhibited a marked reduction in FSP1 expression within the ovaries and oocytes. Pharmacological inhibition of FSP1 disrupted germinal vesicle breakdown and polar body emission, leading to spindle defects and chromosome misalignment. Additionally, FSP1 inhibition persistently activated the spindle assembly checkpoint, resulting in meiotic arrest. At the mechanistic level, inhibition of FSP1 led to an increase in intracellular Fe²⁺ levels, enhanced dihydroethidium fluorescence, excessive accumulation of reactive oxygen species, and intensified lipid peroxidation. Disruptions in ferroptosis-associated gene expression further indicated that oocytes underwent ferroptosis. Moreover, mitochondrial dysfunction was evident following FSP1 inhibition, as reflected by aberrant mitochondrial distribution, diminished ATP production, and an elevated mitochondrial membrane potential. Collectively, these results establish FSP1 as a key regulator of oocyte meiotic maturation by modulating iron homeostasis and mitochondrial function, while its inhibition triggers ferroptosis-dependent meiotic failure.

Background: Uveal melanoma (UVM) seriously affects people's health and quality of life. Here, the mechanism of a disintegrin and metallopeptidase domain 10 (ADAM10) was elucidated in UVM.

Methods: The clinical prognosis and potential biological function of ADAM10 gene in UVM patients were assessed using a series of bioinformatics methods. RT-qPCR and Western blot assay were employed to detect genes expression. Cell apoptosis and viability were examined by flow cytometry, clone formation and CCK-8 assays. The migrated and invasive abilities were analyzed by wound healing and transwell assays. Tumor growth was performed in Xenograft mouse model.

Results: We found that ADAM10 expression was significantly associated with poor prognosis of UVM patients, and its prognostic significance for UVM patients was determined by distinct clinical characteristics. In vitro, ADAM10 expression was upregulated in MUM-2B and C918 UVM cell lines. More importantly, ADAM10 downregulation discouraged cell viability, metastasis but triggered apoptosis of UVM cells. Moreover, ADAM10 upregulation can promote the tumor growth of UVM in vivo. Mechanically, ADAM10 downregulation blocked the Wnt/β-catenin pathway in UVM.

Conclusion: Upregulation of ADAM10 stimulates the malignant behaviors of UVM through activating the Wnt/β-catenin pathway.

Estrogen receptor β (ERβ) plays a pivotal role in regulating intestinal epithelial function and inflammation. Its involvement in inflammatory bowel diseases (IBD), particularly in ulcerative colitis (UC), remains poorly understood, despite emerging evidence pointing to its anti-inflammatory properties. This study investigated ERβ expression in UC patients using quantitative PCR, Western blot, and immunofluorescence. To investigate the functional role of ERβ, a DSS-induced colitis mouse model and LPS-treated HT-29 cells were used. Autophagy activity was evaluated through Western blot, transmission electron microscopy (TEM), and autophagy inhibitors. Co-immunoprecipitation (Co-IP) and dual luciferase reporter assays were employed to explore the interaction between ERβ and hypoxia-inducible factor-1α (HIF-1α), as well as the regulation of ATG-9a expression. The results demonstrated that ERβ expression was significantly downregulated in the inflammatory colons of UC patients. In vivo, ERβ activation by ERB041 alleviated DSS-induced colitis in mice, reducing weight loss, histopathological damage, and inflammatory cytokine levels. In vitro, ERB041 enhanced autophagy in LPS-treated HT-29 cells, accompanied by a reduction in pro-inflammatory cytokines. Furthermore, ERβ activation promoted the expression of tight junction proteins and preserved epithelial barrier integrity. Co-IP and dual luciferase assays revealed that ERβ interacted with HIF-1α and modulated ATG-9a-mediated autophagy. These results indicate that ERβ alleviates intestinal inflammation and activates HIF-1a and ATG-9a-mediated autophagy, providing new insights into the therapeutic potential of targeting ERβ in UC and highlighting its role in maintaining intestinal homeostasis.

Background: Non-alcoholic fatty liver disease (NAFLD), along with non-alcoholic steatohepatitis (NASH), lacks definitive therapy and typically remains asymptomatic until reaching advanced stages. Lipid metabolism and inflammation management using probiotics such as Bifidobacterium adolescentis is suggested to alleviate or suppress NAFLD development. Hence, this study aims to investigate the effects of Bifidobacterium adolescentis treatment on mitigating pyroptosis, an inflammatory cell death pathway, in the liver of rats with NAFLD induced by high-fat diet (HFD) and streptozotocin (STZ) administration.

Methods: Forty 8-week adult male Sprague Dawley rats were divided into four groups. Bifidobacterium adolescentis was administered for 8 and 16 weeks at 4×10¹⁰ CFU/day to rats fed a high-fat diet (HFD). Subsequently, the mRNA expression levels of pyroptotic-related genes including Cas1, Cas3, Cas11, NLRP3, GSDMD, IL-1β, and NF-κB were quantified in liver tissue using quantitative polymerase chain reaction (qPCR). Histopathological alterations and stereological changes in liver structure, as well as lipid profile (FBG, TG, TC, HDL, LDL), and liver indices (ALT, AST, ALP, LDH), were also evaluated across the different groups.

Results: Bifidobacterium adolescentis administration significantly reduced the expression levels of NF-κB and pyroptotic-related genes. Additionally, this probiotic effectively reversed the adverse effects of the high-fat diet (HFD) on liver volume, Kupffer cell numbers, and hepatocyte nuclei. Furthermore, it improved the lipid profile and liver indices of rats fed with the HFD.

Conclusion: This study demonstrates that B. adolescentis supplementation prevents diabetes-induced liver injury by attenuating pyroptosis. These findings suggest that Bifidobacterium adolescentis may be a promising therapeutic approach for managing NAFLD and its associated complications, primarily by modulating key genes associated with pyroptosis and inflammation in rats fed with a high-fat diet.

Our study aimed to elucidate the mechanism by which circTHADA competitively adsorbs miR-494-3p to regulate CASP1-mediated endothelial cell (EC) pyroptosis in diabetic retinopathy (DR). To be specific, we used high glucose (HG)-induced human retinal microvascular endothelial cells (HRMECs) as DR cell models and streptozotocin (STZ)-treated mice as DR mouse models. The expression levels of circTHADA, miR-494-3p, CASP1, NLRP3, GSDMD-N and IL-1β were detected and flow cytrometry was applied to measure cell pyroptosis rate and dual luciferase reporter assays were utilized to determine the direct binding sites. As a result, exacerbated EC pyroptosis in DR was detected in DR cell and mouse models. Based on DE-circRNA profiles by microarray and experimental verification, circTHADA was filtered and identified to regulate CASP1-mediated EC pyroptosis. miR-494-3p was then proven to be involved in circTHADA-mediated ceRNA network by bioinformatics analysis and experimental verification. Further gain- and loss-of-function experiments and rescue experiments revealed the function of the circTHADA/miR-494-3p/CASP1 axis in pyroptosis.

EtOH (Ethanol)-induced cardiotoxicity (EIC) is intimately associated with perturbed lipid metabolism. Lipid droplet-Mitochondria contacts (LD-Mito contacts) are important nodes in lipid metabolism. However, the roles of LD-Mito contacts in EIC have yet to be clarified. In the present study, EtOH exposure induced a significant build-up of LD in cardiomyocytes accompanied by the disturbances in lipogenesis and lipolysis. Upon EtOH treatment, we also observed a substantial decrease in LD-Mito contacts, downregulation of the tethering protein PLIN5 (Perilipin 5), and reduced fatty acid (FA) flux from LD to mitochondria. Overexpression of full-length PLIN5, but not its truncated form (PLIN5Δ), reversed the reduction in LD-Mito contacts and restored FA flux. A synthetic LD-Mito-Linker was generated to exclude the influence of PLIN5's versatile functions and investigate the specific role of LD-Mito contacts in EIC. Tethering LD to mitochondria by the synthetic linker restored the LD-Mito contacts and FA flux in EtOH-treated cardiomyocytes. Inflammation and cardiomyocyte death were measured to indicate lipotoxicity in EIC. Our results demonstrated that overexpression of PLIN5Δ ameliorated EtOH-induced cardiomyocytes death and inflammation whereas restoration of LD-Mito contacts by the synthetic linker aggravated apoptosis, inflammatory response, oxidative stress and Mitochondrial membrane potential depolarization . These findings indicated that loss of LD-Mito contacts and the blocked FA flux may act as a cellular adaptive response to EtOH exposure, thus targeting LD-Mito contacts may serve as a potential therapeutic strategy to combat EIC.

Ovarian cancer (OC) is one of the most fatal gynecological carcinomas, causing significant detriment to women's health. Protein regulator of cytokinesis 1 (PRC1) is a microtubule-associated protein that is found to be highly expressed in many different cancers. Despite this, the exact way in which PRC1 stimulates the growth of OC has yet to be completely understood. Our research demonstrated that PRC1 expression was increased in OC, which was closely related to poor prognosis. Moreover, PRC1 exhibited noteworthy efficacy in enhancing the proliferation and migration capacities of OC cells, as well as affecting the cell cycle in OC cells. Silencing PRC1 significantly suppressed OC growth in vivo. Mechanically, PRC1 could interact with RPL4, which caused a decrease in RPL4/MDM2 complex formation, resulting in the enhanced ubiquitination of p53 and a reduction of p53 proteins. These findings revealed that PRC1 was involved in the RPL4-MDM2-p53 pathway thus playing a tumorigenic role on OC.