The cGAS-STING pathway has been extensively studied as a driver of neuroinflammation in the context of brain aging and neurodegeneration. However, whether cGAS plays a homeostatic role in microglial function under physiological conditions — and how its expression levels evolve at progressive ages in specific brain immune populations — remains poorly understood. Given that microglia represent the predominant cGAS-expressing cell type in the central nervous system, we hypothesized that its constitutive absence may compromise microglial competence and neuro-immune integrity across different ages, independently of overt pathology. Flow cytometric analysis of WT mouse brains revealed that cGAS expression levels in CD45low/CD11bhigh microglia, CD45high/CD11bhigh (macrophages), and CD45high/CD11b− (lymphocytes) cells undergo the most notable transition occurring between 3 and 6 months, suggesting that high cGAS availability may be restricted to a defined window of early adulthood. Western blot and immunofluorescence analyses confirmed a marked reduction in total cGAS and STING protein levels in aged WT brains. In parallel, cGASKO mice exhibited persistent alterations in exploratory behavior across all ages examined, alongside progressive deficits in recognition and spatial memory emerging from adulthood. These behavioral alterations were accompanied by an increased area of neurodegeneration in the somatosensory cortex and hippocampus from 3 months of age. Microglial analysis revealed a consistent reduction in cell density, morphological profile, and reduced Iba1 signal in both brain regions. Flow cytometric profiling showed a sustained downregulation of MHCII, CD86, and CD206 in cGASKO microglia, suggesting functional hyporesponsiveness. Finally, while macrophage infiltration remained unaffected in cortex but reduced in hippocampus, cGASKO mice failed to sustain the age-related increase in lymphocytes population recruitment observed in WT animals, despite showing elevated lymphocyte levels at young age. These findings suggest that cGAS may play an essential homeostatic role in maintaining microglial competence and neuro-immune coordination throughout the lifespan. The early physiological decline of cGAS in brain immune populations, combined with the broad neuroimmune dysfunction observed in cGASKO mice, raises the possibility that this early window of cGAS expression may contribute to the brain's capacity to sustain immune surveillance and resist age-related neurodegeneration. Collectively, these data position cGAS deficiency as a potential model of accelerated microglial aging with implications for understanding the cellular mechanisms underlying brain vulnerability during physiological senescence.
Type 2 diabetes mellitus (T2DM) represents a major and widespread metabolic disorder affecting millions of people worldwide. A key mechanism underlying the onset and progression of T2DM is insulin resistance.The main causes of insulin resistance are a sedentary lifestyle and obesity. Obesity promotes adipose tissue inflammation by increasing immune cell infiltration and inflammatory mediator production, alters gut microbiota toward a pro-inflammatory profile, and induces metabolic endotoxemia, resulting in low-grade systemic inflammation. Together with oxidative stress and elevated free fatty acids, these processes promote insulin resistance, hepatic gluconeogenesis, and pancreatic β-cell dysfunction, thereby exacerbating T2DM. Interestingly, berberine, which is a natural alkaloid with antioxidant and anti-inflammatory properties, affects all of the aforementioned events, thereby potentially contributing to the alleviation of insulin resistance and improvement of T2DM. This article provides a thorough narrative review of the available mechanistic and clinical evidence regarding the potential effects of berberine on T2DM by integrating findings from in vitro, animal, and clinical studies. In addition, this review critically examines the limitations, and translational relevance of the existing evidence. It also discusses the current status of berberine-containing pharmaceutical and nutraceutical formulations and highlights key pharmaceutical and translational challenges. The available evidence indicates that berberine has the potential to influence multiple pathogenic pathways implicated in T2DM and may thereby improve insulin sensitivity. Nevertheless, most mechanistic evidence remains preclinical, and further formulation standardization together with well-designed clinical trials are required to define its efficacy, safety, and optimal dosing before routine clinical use.
Climate change is increasingly threatening the sustainability and productivity of fruit tree systems worldwide. Rising temperatures, altered precipitation patterns, more frequent extreme weather events, and elevated atmospheric CO2 are affecting physiological and phenological processes that regulate tree growth, reproduction, and fruit quality. Key impacts include reduced winter chilling, disrupted dormancy release, irregular flowering, impaired pollination, and altered fruit development, leading to lower yield stability and quality. Due to their long life cycles, fruit trees are particularly vulnerable to cumulative environmental stresses such as heat, drought, salinity, and irregular rainfall, which can impair photosynthesis, growth, and reproductive performance while increasing susceptibility to pests and diseases. This review synthesizes current knowledge on the effects of major climate-related factors, including temperature, water availability, and elevated CO2, on the physiology, phenology, pollination, and productivity of fruit trees. Particular emphasis is placed on the role of genotype diversity and rootstock–scion interactions in improving climate resilience. The review also highlights key adaptation strategies, including the use of climate-resilient cultivars, optimized rootstock selection, efficient water and nutrient management, and climate-smart orchard practices. By integrating physiological, ecological, and production-level responses, this review provides a framework for enhancing the resilience and sustainability of fruit production under changing climatic conditions.
Abstract Background Genetic improvement of rice for biotic stress is a major and continuous breeding objective owing to changing pest and disease scenarios. In response to climate change, an outbreak of new pathotypes and biotypes results in huge yield loss which directly impacts the economic stability and food security. The development of varieties with multiple resistance genes for a particular disease is the most effective approach to the combating mechanism against evolving pathogens and insects. Multiple resistance genes against blast ( Pi54 ), bacterial leaf blight ( xa5 , xa13 , Xa21 ), and brown plant hopper ( Bph17 , Bph3 , bph2 ) were pyramided through marker-assisted forward breeding by attempting multiple crosses involving six parental lines. The F 4 lines were screened and the resistant lines were further reconfirmed at the F 5 stage through precise phenotyping and also by genotyping with trait-specific markers. Finally, the selected lines were evaluated for the agronomic performance. Results Four lines were selected from the population which carrying seven resistance genes against BL, BB, BPH with superior agronomic performance. Another line, x21302-239, harboring all the resistant genes and showing a resistant response in screening experiments for all three stresses, with marginal agronomic performance (single-plant yield: 33 g), can be utilized as a desirable donor to develop elite rice cultivars with multiple biotic stress resistance. Conclusions Homozygous genetic background is more favorable for epidemic outbreak in a short period of time in comparison with the population of multiple genetic background. Since this complex genetic background disturbs the infectious cycle of the pathogen. So that a variety or a hybrid which developed from multiple parental lines are notable for their durable resistance than the monogenic resistance variety.
As a commonly used immunosuppressant, mycophenolate mofetil (MMF) is widely prescribed after organ transplantation because it is not toxic to the liver and kidney. However, its use is associated with certain gastrointestinal toxicity and increased risk of opportunistic infections. Meanwhile, whether it affects vascular development is unclear. Herein, a zebrafish model was utilized to evaluate whether MMF affects vascular development, and the results showed that MMF exhibited teratogenicity, shortened the body length, caused pericardial edema, and delayed yolk sac absorption in zebrafish. In addition, although Intersegmental vessels showed hyperbranching, opposite changes were observed in cerebrovascular, with cerebrovascular inhibition. Moreover, MMF exposure upregulated oxidative stress levels, inhibited notch signaling and mitogen-activated protein kinase (MAPK) signaling, and affected the expression of vascular endothelial growth factor and receptor tyrosine kinase. Treatment with the antioxidant astaxanthin and the notch signaling pathway activator sodium valproate did not rescue the vascular development defects, indicating that MMF affects multiple signal pathways. However, the treatment of MMF-exposed zebrafish with guanosine and the MAPK signaling activators diprovocim significantly rescued the angiogenesis-related defects, suggesting that MMF induces ISV hyperbranching in zebrafish by inhibiting MAPK signaling and inosine 5′-monophosphate dehydrogenase (IMPDH).
Telomeres play a crucial role in maintaining genomic stability in healthy cells. However, they gradually shorten during the cell cycle, leading to chromosomal instability. Telomere length and telomerase activity are vital factors that counteract cellular degradation in cancer development and tumor persistence. Telomerase, which is activated in most cancer cells due to telomerase catalytic subunit (hTERT) overexpression, serves as a universal biomarker that is essential for cancer cell growth and survival. The upregulation of hTERT, often associated with G > A mutations in its promoter region, is frequently implicated in cancer progression. Consequently, anti-telomerase therapy has been proposed as a potentially more efficacious alternative to conventional treatment. Small-molecule inhibitors have garnered significant attention owing to their selectivity or ability to modulate multiple proteins. However, challenges, such as low response rates, brief response durations, toxicity, and resistance persist. Several strategies have been proposed to target telomerase activity and the telomere structure. These include the utilization of G-quadruplex-stabilizing compounds and telomere-specific oligonucleotide inhibitors of telomerase such as GRN163L and T-oligos. Another therapeutic approach involves the use of biological antisense oligonucleotides that specifically inhibit hTERT and human telomerase RNA component genes, potentially reducing telomerase activity and generating robust DNA signals in cancer cells. Immunotherapy targeting hTERT represents a recent advancement in cancer treatments. This approach leverages the immune system to target cancer cells with high hTERT expression, thereby offering a potentially more reliable treatment strategy. This review provides an overview of current research on telomerase-targeting small-molecule inhibitors, antisense oligonucleotides, and immunotherapy, discussing their mechanisms, clinical applications, and prospects in cancer treatment.
Telomeres play a crucial role in maintaining genomic stability in healthy cells. However, they gradually shorten during the cell cycle, leading to chromosomal instability. Telomere length and telomerase activity are vital factors that counteract cellular degradation in cancer development and tumor persistence. Telomerase, which is activated in most cancer cells due to telomerase catalytic subunit (hTERT) overexpression, serves as a universal biomarker that is essential for cancer cell growth and survival. The upregulation of hTERT, often associated with G > A mutations in its promoter region, is frequently implicated in cancer progression. Consequently, anti-telomerase therapy has been proposed as a potentially more efficacious alternative to conventional treatment. Small-molecule inhibitors have garnered significant attention owing to their selectivity or ability to modulate multiple proteins. However, challenges, such as low response rates, brief response durations, toxicity, and resistance persist. Several strategies have been proposed to target telomerase activity and the telomere structure. These include the utilization of G-quadruplex-stabilizing compounds and telomere-specific oligonucleotide inhibitors of telomerase such as GRN163L and T-oligos. Another therapeutic approach involves the use of biological antisense oligonucleotides that specifically inhibit hTERT and human telomerase RNA component genes, potentially reducing telomerase activity and generating robust DNA signals in cancer cells. Immunotherapy targeting hTERT represents a recent advancement in cancer treatments. This approach leverages the immune system to target cancer cells with high hTERT expression, thereby offering a potentially more reliable treatment strategy. This review provides an overview of current research on telomerase-targeting small-molecule inhibitors, antisense oligonucleotides, and immunotherapy, discussing their mechanisms, clinical applications, and prospects in cancer treatment.
The reliability and accuracy of relative quantification of transcripts through quantitative real-time PCR (qPCR) is dependent on the use of stable reference genes for normalization. However, the expression stability of various reference genes may differ across the tissues and patho-physiological conditions. The present study investigated the stability of multiple candidate reference genes in Japanese quail (Coturnix japonica) embryos, subjected to embryonic thermal conditioning (ETC). Fertilized quail eggs (n = 200) were incubated under standard conditions and heat exposure (ETC) of incubated eggs was undertaken at 39.5 °C for 10 h daily in the embryonic days 6–8 and 12–14, maintaining constant humidity (55
The targeted, substrate-specific degradation of paternal mitochondria inside the zygote, known as post-fertilization sperm mitophagy, is a crucial and evolutionarily conserved early embryonic event. It ensures the exclusive maternal inheritance of the mitochondrial genome. Post-fertilization sperm mitophagy was initially thought to only be achieved via the ubiquitin–proteasome system. Until pro-autophagic receptor proteins such as SQSTM1, GABARAP, as well as the proteasome-interacting ubiquitinated protein dislocase VCP, were identified as contributors to the degradation of the sperm mitochondria early after mammalian fertilization. This synergy of proteasomal and autophagic pathways ensures a timely degradation of sperm mitochondria shortly after fertilization. The discovery of these autophagic receptors lead researchers to believe there might be other autophagic receptors and determinants necessary for proper post-fertilization sperm mitophagy. Based on the established inventory of proteins from mass spectrometry trials of boar spermatozoa exposed to porcine oocyte extracts in an intra-specific porcine cell-free system (CFS), five candidate mitophagy determinants were further investigated in this study, namely LACTB, PRDX3, PSMA8, TOMM34, and FUNDC1. These proteins of interest were studied and validated by using in vitro fertilization (IVF) protocols, cell imaging of spermatids, spermatozoa, oocytes and zygotes, protein interactome analysis, and the porcine CFS. The proteins PSMA8 and TOMM34 behaved in accordance with our proteomic study predictions. The PSMA8 labeling increased after exposure to CFS; in agreement with the classification PSMA8 was given from the mass spectrometry findings. TOMM34 underwent a visible decrease in labeling after exposure to CFS, which also agreed with its proteomic classification; this labeling persisted in IVF zygotes. Except for LACTB, the examined proteins showed mutual interactions as well as interactions with previously identified sperm mitophagy factors in the STRING interactome analysis. Results from this study validate the novel porcine CFS as a valuable tool for the exploration of early fertilization events at a molecular level. Future phenotyping and functional studies using porcine CFS will advance the understanding of mitochondrial inheritance and zygotic development and potentially shed light on the origins of certain mitochondrial diseases arising from the failure of post-fertilization sperm mitophagy.
Salvadora persica (miswak) has many useful biological activities. Some studies have reported that miswak can be used as a contraceptive agent. However, no available investigations explain the histomorphological structure of ovarian follicles after miswak aqueous extract (MAE) administration. Twelve female Wistar albino rats were divided into two equal groups. In the control group (CG), the animals received normal saline daily for 4 weeks. While in the miswak treated group (MTG), the animals received orally 900 mg/kg of body weight of the MAE daily for the same period. At the end of the experiment, the rats were anesthetized and then euthanized by cervical dislocation. The ovaries were dissected, removed, weighted, fixed, and processed for histological examination by light and electron microscopy. Results revealed that the ovaries of CG showed various stages of follicular development. While in the MTG, the ovaries exhibited follicular atresia. The immunoexpression of caspase-3, progesterone receptors (PR), and estrogen receptors alpha (ERA) in the MTG and CG were reported. Our preliminary animal research indicates that MAE can inhibit follicular development and induce follicular atresia. These findings suggest a potential antifollicular and/or antiovulatory effect that requires confirmation through dose-response, fertility, toxicity, and clinical studies.
Abstract Sour orange ( Citrus aurantium L.) is a hybrid species of considerable agronomic, medicinal, and industrial importance; however, its wild genetic resources remain largely underexplored. This study evaluated 24 naturally occurring wild sour orange genotypes collected from the Hassa district of Hatay, Türkiye, using an integrated framework combining morphological, pomological, biochemical, and ISSR molecular markers. The combined use of multi-trait phenotypic measurements and molecular markers enabled a comprehensive assessment of genetic and phenotypic diversity within this localized germplasm. Substantial variation was observed in fruit and leaf traits, indicating strong potential for the selection of contrasting ideotypes. Biochemical analyses revealed marked diversity in total phenolic and flavonoid contents, antioxidant capacity, and vitamin C levels, allowing the identification of genotypes that combine high nutritional quality with commercially acceptable fruit size. ISSR analysis produced a high polymorphism rate (75.72%), confirming pronounced genetic diversity within this micro-region. Multivariate analyses, including correlation, regression, principal component analysis, and hierarchical clustering visualized through heat maps, clearly discriminated genotypes according to fruit size, seed productivity, and phytochemical composition. Overall, the results highlight the presence of considerable genetic diversity within the Hassa district; however, given the geographically restricted sampling, these findings should be interpreted as representative of a localized germplasm rather than the entire species distribution. The study provides a practical basis for selecting superior genotypes for rootstock breeding, fresh fruit utilization, and nutraceutical applications.
Abstract Background Endometrial cancer (EC) is a common malignancy of the female reproductive tract worldwide. While comprehensive genomic analyses have identified multiple genetic alterations in EC, disease models remain relatively rare, limiting the ability to conduct disease studies. Establishing a hybrid in vitro and in vivo model of EC and comparing it to its in vivo counterpart could greatly advance research into EC mechanisms and treatment strategies. Methods We established genetically engineered EC mice ( Pten flox/+ ; Stk11 flox/flox ; LTF-Cre (PSC)) and constructed a corresponding epithelial cell line from isolated uterine tumor tissues using conditional reprogramming techniques. The cell line was successfully passaged, frozen, and stored in a biobank, providing a valuable resource for future EC research. Results The PSC-derived cell line stably retained the mutant alleles of Pten and Stk11 during passaging, preserving the genetic profile of the original tumor. In addition, we performed various functional assays on isolated mouse cell linesand formed tumors after subcutaneous implantation of cells in immunodeficient mice and C57BL/6 mice with an intact immune system. Hematoxylin and eosin (H&E) staining and immunohistochemical staining were used to characterize the histopathology and biomarkers of the subcutaneously implanted tumors, which were highly similar to those of the parental mouse uterine tumors. Conclusions The isolated mouse cell line provides a powerful platform for basic and translational EC studies, matching well with the corresponding in vivo models. The fidelity of the cell line to human-relevant genetic alterations and tumor biology makes it a valuable tool for investigating EC pathogenesis and evaluating novel therapeutic approaches.
Abstract Background Abnormal syncytialization of cytotrophoblast (CTB) cells is known to be associated with preeclampsia (PE), however, the underlying molecular mechanisms remain elusive. Results In this study, proteomic analysis of placental tissues was performed to identify proteins involved in PE pathogenesis. NCK Adaptor Protein 1 (NCK1) was found to be downregulated in placental tissues from patients with early-onset PE. In vitro functional assays revealed that NCK1 was markedly increased during spontaneous syncytialization of CTB cells. NCK1 overexpression alleviated hypoxia-induced defects in CTB cell fusion. Mechanistically, NCK1 upregulated both the expression and stability of Peroxisome Proliferator-Activated Receptor Gamma (PPARG), accompanied by increased m6A modification of its mRNA. Overexpression of PPARG significantly rescued hypoxia-induced syncytialization impairment, whereas PPARG deficiency abolished the promotive effects of NCK1 on CTB syncytialization. Additionally, phosphorylation of Eukaryotic Translation Initiation Factor 3 Subunit (EIF3D) was significantly increased during spontaneous syncytialization, and NCK1 overexpression partially reversed hypoxia-induced suppression of EIF3D phosphorylation. EIF3D knockdown significantly reduced m6A levels on PPARG mRNA, likely due to impaired NCK1-mediated suppression of Alpha-Ketoglutarate-Dependent Homolog 5 (ALKBH5), an m6A demethylase. Furthermore, NCK1 enhanced EIF3D phosphorylation by inhibiting its O-GlcNAcylation. Conclusions Our findings demonstrate that NCK1 stabilizes PPARG by modulating the crosstalk between O-GlcNAcylation and phosphorylation of EIF3D, thereby restoring hypoxia-impaired syncytialization in CTB cells. This study identifies the NCK1/EIF3D/PPARG axis as a molecular pathway potentially relevant to trophoblast fusion dysfunction in PE.
Abstract Background The incidence of erectile dysfunction (ED) rises sharply with age, which has brought a huge social burden. The exploration of aging-related ED is hindered due to the limitations of natural aging models. As an ideal inducer of accelerated aging models, the mechanism of D -galactose ( D -gal) in ED has not been fully elucidated. Therefore, this study aimed to verify the feasibility of aging-related ED induced by D -gal and explore its potential mechanism. Methods Primary corpus cavernosum smooth muscle cells (CCSMCs) and human umbilical vein endothelial cells (HUVECs) were incubated with concentration gradients of D -gal. Meanwhile, aging rat models were established via intraperitoneal and subcutaneous injection of D -gal, and the erectile function of all subjects was measured by electrical stimulation. Samples from both cells and animals were then collected for subsequent detection. Results Cellular senescence was confirmed in both cells and was accompanied by oxidative stress, mitochondrial dysfunction, and cell cycle arrest in vitro. The erectile function of rats treated with D -gal was affirmed to be impaired compared with normal rats. In addition to the above-mentioned pathological alterations, tissue fibrosis was also associated with D -gal administration in vivo. Moreover, no significant difference was found between the two administration methods in inducing senescence of rats. Conclusions D -gal could promote the progression of aging-related ED, accompanied by the exacerbation of oxidative stress, mitochondrial dysfunction, cell cycle arrest, and fibrosis. Our data suggested that D -gal may be an ideal mediator for the induced model of aging-related ED, which could facilitate preclinical exploration of this disease.
Ovarian theca cells constitute essential components of the follicular microenvironment and play central roles in follicular development, steroidogenesis, and endocrine regulation. Despite their significance, the developmental origins, differentiation processes, and functional dynamics of theca cells remain incompletely defined, particularly in humans. This review provides an updated synthesis of current knowledge on the ontogeny, molecular signaling pathways, and intercellular interactions of theca cells. It also presents recent findings on the role of theca stem or progenitor cells and their relevance to reproductive disorders, including polycystic ovary syndrome (PCOS), hyperthecosis, and ovarian insufficiency. A comprehensive literature search was conducted using PubMed, Scopus, and Web of Science through May 2025. Search terms included "theca cells," "theca progenitors," "follicular development," "ovarian differentiation," "steroidogenesis," and "reproductive disorders." Original research articles and reviews providing mechanistic insights were included. Current evidence indicates that several signaling pathways, including IGF, SCF, FGF, members of the TGF-β superfamily, GDF9, BMPs, and Hedgehog proteins, are involved in the recruitment, proliferation, and differentiation of theca cells. Bidirectional communication among oocytes, granulosa cells, and theca cells remains essential for folliculogenesis. Increasing evidence links theca cell dysfunction to the pathophysiology of PCOS, primary ovarian insufficiency, and reproductive aging. The identification of theca stem cells (TSCs) and their proposed roles in ovarian regeneration represents an important conceptual advance. A deeper understanding of theca cell biology may guide the development of targeted strategies for infertility and ovarian dysfunction. In contrast, the integration of TSC biology offers new directions for reproductive medicine and reproductive health.
Mesenchymal stem cells (MSCs) hold great potential for regenerative medicine and tissue engineering. However, their clinical use is limited by low in vivo availability and rapid senescence during ex vivo expansion, posing a major challenge for scaled production. Growing evidence indicates that microenvironmental oxygen tension critically regulates MSCs fate. This study aims to systematically evaluate how different oxygen tensions modulate MSCs aging and function, providing evidence to optimize MSCs expansion protocols for clinical applications. Human umbilical cord derived-mesenchymal stem cells (hUC-MSCs) were maintained under normoxic (20
Abstract Plums ( Prunus spp.) are among the most diverse fruit tree species, yet their wild relatives remain poorly characterized despite their importance for breeding, conservation, and understanding evolutionary diversity. Here, we investigated a wild plum population from Mount Erciyes, located in Central Anatolia, Türkiye, through integrated morphological, textural, total content-based biochemical, and plastome analyses. Floral morphology of the Erciyes wild plums displayed intermediate features between apricot ( P. armeniaca ) and cultivated plum ( P. domestica ), while fruit texture exhibited significantly higher peel hardness and flesh resistance compared with the reference cultivar ‘Papaz Eriği’. Illumina sequencing of the chloroplast genome produced a 157,840 bp plastome with a conserved quadripartite structure, encoding 125 genes. Comparative analyses revealed minor structural differences in the LSC and SSC regions, whereas the IR regions remained highly conserved. Phylogenetic reconstruction based on complete plastomes clustered the Erciyes wild plums with P. domestica and P. cerasifera , reflecting plastome-level phylogenetic affinity. This study identified significant phenotypic and total content-based biochemical diversity in wild plum genotypes. Key traits such as fruit and seed weight, color, and bioactive compound contents showed wide variability. Based on the heatmap hierarchical clustering analysis, the genotypes and the examined traits were divided into two main groups exhibiting significant positive and negative correlations among the traits. Together, these findings provide a characterization of a previously undocumented wild plum from Mount Erciyes and establish a baseline reference for future studies on plum evolution, conservation, and breeding.
Inflammatory bowel disease (IBD) is a chronic inflammatory disease caused mainly by immune overactivation and intestinal mucosal barrier disruption. Coffea arabica pulp aqueous extract (CPE) contains a number of bioactive phenolic compounds that exhibit antioxidant and anti-inflammatory effects with previously unexplored application in experimental colitis. The aim of this study is to determine whether CPE produces anti-colitogenic effect and its possible mechanism of action. We found that CPE attenuated all IBD-related phenotypes and increased survival rates in dextran sulfate sodium (DSS)-induced colitis mice. In addition, CPE significantly suppressed mRNA and protein expression of myosin light-chain kinase (MLCK). Furthermore, CPE also inhibited MLCK recruitment to apical junction of colonic tissues of colitis mice. Although CPE had no effect on mRNA expression of tight junction genes, it reversed inflammation-mediated downregulation of ZO-1, occludin, and claudin-4 proteins in colitis mice. Importantly, CPE was able to recover ZO-1 and occludin localization to apical junction and suppressed tight junction-dependent leak pathway permeability in colitis mice. Indeed, CPE was also capable of stimulating sirtuin-1 (SIRT-1) in colonic tissues obtained from DSS-induced colitis mice, which is known to suppress inflammation and enhance intestinal barrier function. Therefore, SIRT-1 has been shown to be associated with CPE treatment in IBD model.
BACKGROUND:Filamin A (FLNA) is an actin-binding protein that regulates mechanosensitivity and functions as an intracellular signaling scaffold in various cell types. It has also been implicated in tumor growth. We recently reported FLNA expression in human ovarian granulosa cells and in KGN cells, a granulosa cell tumor (GCT) line. RESULTS:Immunohistochemistry analysis of 51 GCT samples revealed heterogeneous FLNA expression, with approximately 20% showing weak, 18% strong, and the majority moderate expression. We therefore conducted functional studies in KGN cells using CRISPR/Cas9 gene editing. A proteomic approach revealed marked changes in protein abundance upon FLNA depletion: proteins with increased abundance were predominantly related to adhesion, cytoskeletal organization, regulation of cell shape, and lipid metabolic process, whereas those with decreased abundance were associated with DNA replication, cell division, and cell cycle regulation. FLNA-knockout cells showed enlarged cell sizes, reduced proliferation, and slightly affected steroidogenesis. Disruption of FLNA further reduced migration velocity, altered actin cytoskeletal alignment under flow, and modified expression of genes involved in cytoskeletal architecture, adhesion, and mechanosensing under shear stress. CONCLUSIONS:Our results identify crucial roles of FLNA in shaping the cellular architecture, motility, and proliferation of KGN cells. Consequently, alterations in FLNA expression may influence intracellular signaling, and responsiveness to mechanical cues in both physiological and pathological contexts.
Abstract Background Follicular disorders, often driven by ROS-induced granulosa cell apoptosis, are a major cause of female infertility. While α-ketoglutarate (AKG), also known as 2-oxoglutarate, can improve follicular development, the underlying mechanisms remain unclear. Given that AKG is the primary substrate of oxoglutarate dehydrogenase (OGDH), this study aimed to investigate how OGDH mediates the protective role of AKG against oxidative stress and in supporting follicular development. Result AKG treatment advanced puberty onset and increased the number of corpora lutea in mice. It alleviated oxidative stress and apoptosis in granulosa cells by upregulating CAT and downregulating P53. Crucially, OGDH physically interacted with CAT and SOD2 and boosted their enzymatic activities, thereby reinforcing AKG’s antioxidative effects. Knockdown of OGDH markedly impaired the ability of AKG to promote follicular development. Conclusions These findings identify OGDH as a key mediator of AKG’s protective role in follicular development through modulation of oxidative stress and apoptosis. This work provides mechanistic insight into AKG function and supports its potential as a therapeutic strategy for follicular disorders.
