Search Results (627)

Background: Oxidative stress, neuroinflammation, impaired autophagy, and mitochondrial dysfunction are major contributors to ageing and neurodegenerative diseases. This study investigated whether AgeViva could counteract these processes by modulating redox balance, inflammation, autophagy, and mitochondrial function. Methods: The effects of AgeViva were evaluated using two complementary experimental models: lipopolysaccharide (LPS)-stimulated BV2 microglial cells and Caenorhabditis elegans (C. elegans) nematodes. In BV2 cells, the expression of inflammatory, autophagy-related, and antioxidant response genes, including NRF2, SOD, and GPX, was evaluated by RT-qPCR, while cell viability was assessed by Trypan Blue exclusion assay. In C. elegans, lifespan, healthspan parameters, ROS accumulation, mitochondrial integrity, membrane potential, and the expression of stress-response, longevity, and autophagy-related genes were analyzed following AgeViva supplementation. Results: In LPS-stimulated BV2 cells, AgeViva significantly reduced the expression of mRNA the pro-inflammatory cytokines Interleukin-1 beta (IL-1β) and Tumor Necrosis Factor alpha (TNF-α) while increasing Interleukin-10 (IL-10) levels, AgeViva also induced changes in autophagy-related transcripts, such as modulation of microtubule-associated protein 1a/1b-Light Chain (LC3) and Sequestosome 1 (p62) expression, activated antioxidant-related gene expression, increasing the expression of Superoxide Dismutase 1(SOD1) and Glutathione Peroxidase (GPX). In C. elegans, AgeViva supplementation extended lifespan and improved healthspan parameters, including locomotor activity and pharyngeal pumping. Treated nematodes showed reduced cytosolic and mitochondrial ROS accumulation, preservation of mitochondrial network integrity, and maintenance of mitochondrial-associated fluorescence, reflecting mitochondrial content and/or membrane potential during ageing. Molecular analyses revealed modulation of key pathways involved in stress resistance and longevity, including Insulin-like Growth Factor 1 (Insulin/IGF-1) signaling Dauer Formation-2 and 16 (DAF-2/DAF-16), Skinhead-1 (SKN-1/Nrf2) signaling, and autophagy-related genes, like Ligating (lgg-1), Autophagy-Related-7 (atg-7), Autophagy Related-18 (atg-18), uncoordinated-51 (unc-51), and ectopic p-granules autophagy protein 5 (epg-5). Conclusions: AgeViva promotes healthy ageing by modulating oxidative stress, inflammation, autophagy, and mitochondrial homeostasis. Full article

Red algae (Rhodophyta) are an ancient lineage of photosynthetic eukaryotes that play important roles in marine ecosystems. However, genomic information for many species within the order Rhodymeniales remains limited, particularly for mitochondrial genomes. In this study, we sequenced, assembled, and analyzed the complete mitochondrial genome (mitogenome) of Rhodymenia intricata to investigate its genome organization, gene content, and phylogenetic position within Rhodymeniales. The mitogenome of R. intricata is a circular DNA molecule of 26,213 bp containing 49 genes, including 25 protein-coding genes (PCGs), 21 tRNA genes, and 3 rRNA genes. The genome shows a strong A + T bias (70.9%) and positive AT and GC skews, typical of red algal mitogenomes. Comparative analysis with other Rhodymeniales mitogenomes revealed generally conserved gene content and organization, with several lineage-specific features such as the presence of the rpl20 gene, an additional open reading frame (orf148), and three rRNA genes (rnl, rns, and rns5). Codon usage analysis indicated a preference for leucine and isoleucine codons and dominant start and stop codons (ATG and TAA). Phylogenetic analysis based on a concatenated dataset of 23 mitochondrial PCGs strongly supported the monophyly of Rhodymeniales and confirmed the close relationship between R. intricata and R. pseudopalmata. Overall, this study presents the first complete mitogenome of R. intricata and expands mitogenomic resources for Rhodymeniales, providing new insights into mitogenome evolution and phylogenetic relationships in red algae. Full article

Cumulus expansion is important for oocyte maturation, but its molecular regulation in yak remains unclear. This study examined the association of Yes-associated protein 1 (YAP1) with cumulus expansion-related and autophagy-related molecular markers in cumulus cells isolated from yak cumulus–oocyte complexes collected from ovarian follicles. YAP1 was overexpressed or knocked down, and YAP1-overexpressing cultures were additionally treated with 3-methyladenine (3-MA). Transcript abundance was assessed by RT-qPCR and interpreted descriptively, whereas protein abundance was evaluated by Western blotting; representative immunofluorescence and mCherry-GFP-LC3B images were also examined. YAP1 overexpression significantly increased HAS2 and PTGS2 protein abundance, while PTX3 and TNFAIP6 showed non-significant upward trends; YAP1 knockdown significantly reduced all four proteins. YAP1 manipulation was also associated with changes in Beclin-1, ATG5, p62, and the LC3B-II/LC3B-I ratio. Compared with YAP1 overexpression alone, 3-MA co-treatment significantly reduced HAS2, PTGS2, PTX3, TNFAIP6, Beclin-1, and ATG5, increased p62, and numerically reduced the LC3B-II/LC3B-I ratio without statistical significance. These findings support an association between YAP1 and cumulus expansion-related molecular programs and are consistent with partial involvement of 3-MA-sensitive, autophagy-associated signaling. Direct functional assays and pathway-specific validation are required to establish causality. Full article

Postmenopausal ovarian hormone deficiency (OHD) contributes to overactive bladder (OAB) through oxidative stress, mitochondrial dysfunction, and dysregulated estrogen receptor (ER) signaling. This study investigated whether estradiol (E2) alleviates dysfunction by modulating the ER/Smad/miRNA-200/KLF4/mTOR signaling pathway to restore mitochondrial quality control. Thirty female Sprague-Dawley rats were initially allocated to Sham, Ovariectomy (OVX) and OVX + E2 group; after attrition during the 12-month protocol, six surviving animals per group were included in the principal analyses. The OVX + E2 group received daily intramuscular E2 (IM, 30 μg/kg/day) for one month. Bladder function was assessed via micturition volume and frequency by metabolic cages, cystometrograms, and contractility assays. Mechanisms were analyzed using immunofluorescence, Western blotting, transmission electron microscopy (TEM), and miRNA sequencing. OVX rats exhibited significant overactivity and compromised contractility, accompanied by upregulated ERα and downregulated ERβ/GPER. Molecularly, OHD was associated with the upregulation of the miRNA-200 family (miRNA-200a-3p, miRNA-200b-3p and miRNA-200b-5p), which was accompanied by reduced levels of KLF4 and autophagy proteins (ATG7, ATG12 and Beclin-1), as well as elevated p-mTOR expression. TEM revealed the accumulation of damaged mitochondria with ultrastructural features associated with impaired mitophagy. However, E2 treatment ameliorated these abnormalities. These improvements were associated with the restoration of TGF-β/Smad signaling, downregulation of selected miRNA-200 family members, recovery of KLF4 level, and increased expression of autophagy-related markers, accompanied by improved mitochondrial ultrastructural integrity. This cellular restoration might be correlated with improved urodynamic parameters. E2 might exert therapeutic effects by improving mitochondrial quality, potentially via the ER/Smad/miRNA-200/KLF4 signaling pathway. These findings provided mechanistic insights into estrogen-mediated protection and highlight this proposed signaling pathway as a therapeutic target for postmenopausal OAB. Full article

Mechanical stress is thought to be involved in the pathogenesis and pathophysiology of osteoarthritis (OA). However, much remains to be elucidated regarding how chondrocytes sense and respond to mechanical stress (stress sensing and response factors). Additionally, it still remains unclear whether there are defensive responses and mechanisms to protect against pathological agents and mechanical stress in articular cartilage tissue. This study was designed to determine whether repetitive mechanical force, at physiologic levels, affects the expression of factors regulating autophagy such as the autophagy-related proteins ATG5, Beclin-1, and Parkin, and the autophagy process as well as cellular activity in cultured chondrocytes. Three-dimensional cultured tissue was generated from human chondrocytes using a collagen sponge scaffold. After physiological mechanical loading of the 3D cell–collagen sponge construct, comparative analyses of expression levels of ATG5, Beclin-1, and Parkin were performed in human chondrocytes. Chondrocyte activity and Transmission Electron Microscopy (TEM) analysis for detecting autophagy process were also analyzed with or without repetitive compressive loading. In chondrocytes, 60 min or 180 min repetitive compressive loading significantly decreased the expression of ATG5, Beclin-1 and Parkin in comparison with the non-loading group. TEM analysis indicated that, in normal chondrocytes of the non-loading group, the autophagy process was shown to be progressing. In contrast, repetitive loading decreased the number of autophagosomes and autolysosomes in chondrocytes. In addition, numerous degenerated organelles that had not undergone autophagy were observed within the chondrocytes under repetitive loading. The ATG5 and Beclin-1 proteins are known to play crucial roles in regulating cellular autophagy. Furthermore, repetitive mechanical loading caused a decreased expression of Parkin, a mitophagy regulator in chondrocytes. Our results indicate for the first time that a decrease in mitophagy, as well as cellular autophagy, in response to mechanical stress, even at the physiologic level, leads to the accumulation of defective mitochondria and abnormal cellular proteins, resulting in reduced chondrocyte activity and affecting the maintenance of cartilage tissue homeostasis, ultimately contributing to the progression of OA. Full article

β-propeller protein-associated neurodegeneration (BPAN) is the most prevalent subtype of neurodegeneration with brain iron accumulation (NBIA) and is caused by mutations in the WDR45 gene encoding the autophagy-related protein WIPI4. However, many WDR45 missense variants remain classified as variants of uncertain significance (VUS), highlighting the need for reliable functional assays to support their clinical interpretation. In this study, we identified the yeast ortholog of WDR45 and established a Saccharomyces cerevisiae-based functional complementation assay to assess the pathogenicity of WDR45 variants. We first showed that deletion of the β-propellers that bind polyphosphoinositides (PROPPIN)-encoding genes ATG18 or HSV2 causes mitochondrial dysfunction and impaired respiratory growth. Human WDR45/WIPI4 specifically rescued the respiratory defect of the atg18Δ strain, whereas WDR45B/WIPI3 complemented the hsv2Δ phenotype, establishing yeast Atg18 as the closest functional counterpart of WIPI4, thus clarifying PROPPIN orthology. We then evaluated a panel of WDR45 variants and found that benign variants restored normal growth, whereas truncating and pathogenic missense variants failed to complement the atg18Δ phenotype, validating the assay for functional variant classification. Finally, we analyzed several VUS identified in patients with clinically compatible BPAN and obtained functional evidence supporting their pathogenicity. Overall, our study establishes a simple, robust, and scalable yeast model that enables functional interpretation of WDR45 variants and improves molecular diagnosis of BPAN. Full article

Identifying physiological changes during the transition period is essential for improving the health and productivity of dairy goats. This study evaluated hematological, biochemical, hormonal, oxidative stress, and molecular alterations in Shami goats during the pre-pregnancy, late pregnancy, and early lactation periods. Eighty clinically healthy goats were examined, and blood samples were analyzed for hematological indices, metabolic and hormonal profiles, oxidative stress biomarkers, and relative expression of genes associated with energy metabolism, antioxidant defense, inflammation, and autophagy. Late pregnancy was characterized by significant (p < 0.05) increases in red blood cell count (RBCs), hemoglobin concentration (Hb), neutrophils, albumin, globulin, urea, insulin-like growth factor-1 (IGF-I), and malondialdehyde (MDA), accompanied by decreased glucose, cholesterol, total protein (TP), antioxidant markers, total leukocyte count, packed cell volume, and monocytes. Early lactation was associated with higher non-esterified fatty acid, triiodothyronine (T3), and thyroxine (T4) levels. Genes involved in lipid mobilization and oxidation, ketogenesis, inflammation, cellular stress, and autophagy; sirtuin 1 (SIRT1), peroxisome proliferator-activated receptor alpha (PPARA), carnitine palmitoyltransferase 1a (CPT1A), 3-hydroxy-3-methylglutaryl-coenzyme a synthase 2 (HMGCS2), cluster of differentiation 36 (CD36), lipase E (LIPE), protein kinase amp-activated catalytic subunit alpha 1 (PRKAA1), solute carrier family 2 member 1 (SLC2A1), haptoglobin (HP), interleukin 6 (IL6), heat shock protein 70 (HSP70), heme oxygenase 1 (HMOX1), beclin 1 (BECN1), and autophagy-related protein 5 (ATG5) were significantly upregulated, whereas antioxidant- and glucose transport-related genes nuclear factor erythroid 2-related factor 2 (Nrf2), glutathione peroxidase 1 (GPX1), catalase (CAT), thioredoxin (TXN), and solute carrier family 2 member 4 (SLC2A4) were downregulated during the transition period. The results indicate well-orchestrated metabolic and molecular adaptations that can be employed as biological indicators to track the physiological status of Shami goats. These findings fulfilled the study objective and identified potential biomarkers of the transition period in Shami goats. Full article

The chemical composition of donkey and sheep milk showed significant differences, with donkey milk containing less fat and protein and a higher level of lactose, while sheep milk had more dry matter and a higher proportion of casein and saturated, unsaturated, monounsaturated, and polyunsaturated fatty acids. This study investigated the biological effects of in vitro-digested donkey and sheep milk, and their blends at different ratios, on differentiated human intestinal Caco-2 cells. Gene expression analysis showed that treatment with digested donkey milk (100%) significantly altered the expression of the autophagy-related gene (SQSTM1), tight junction (CLDN4), and innate defense genes (DEFB1, MUC2, and MUC5), showing a non-cytotoxic response and a profile consistent with a barrier-related transcriptional response. In contrast, sheep milk (100%) and certain milk blends (70% and 50% sheep’s milk) down-regulated genes involved in the autophagy process (ULK1, AMBRA, BECN1, ATG5, GABARAP, and SQSTM1), tight junction genes (OCLN and CDH1), and innate defense genes (DEFB1 and MUC2), suggesting a distinct epithelial response that warrants further confirmation. Metabolomic profiling revealed clear compositional and functional distinctions among the digested milks, identifying 166 metabolites that were differentially regulated (136 compounds were down-regulated and 30 up-regulated). Donkey milk digestion yielded a metabolite profile enriched in polar peptides and compounds with reported antioxidant associations, whereas sheep milk digestion showed a different pattern dominated by hydrophobic peptides and lipid-related metabolites. These findings indicate that, under the tested in vitro conditions, donkey milk digestion is associated with gene expression and metabolomic patterns consistent with epithelial barrier support. Full article

Background/Objectives: CRISPR/Cas9-mediated genome editing enables efficient generation of knockout mouse models; however, frameshift mutations do not always result in complete loss of function. The factors influencing functional inactivation following frameshift mutations remain incompletely understood. Here, we tested whether frameshift-dominant targeting of exon 1 is sufficient to generate a null allele of Fgf10, a gene essential for limb formation. Methods: Guide RNAs (gRNAs) were selected using a machine learning-based pipeline to favor microhomology-mediated end joining (MMEJ)-dominant repair. Editing efficiency and indel profiles were assessed via amplicon sequencing in mouse embryonic stem cells (mESCs) and preimplantation embryos. Edited embryos were transferred to surrogate females and analyzed at embryonic day 15.5 (E15.5). Results: Amplicon sequencing confirmed >97% editing efficiency and >80% frameshift alleles in both mESCs and preimplantation embryos, with a predominant 7 bp deletion. Despite highly efficient frameshift-dominant editing, most of the E15.5 embryos were morphologically normal, indicating that exon 1 targeting did not reliably produce null phenotypes. In silico analysis suggested the possible presence of alternative downstream translation initiation sites, and our secretion assay supported this possibility. Initiation from a downstream ATG in a +2 reading frame (e.g., 7 bp deletion) may restore the downstream coding sequence and partially preserve protein function. Conclusions: Frameshift mutations in exon 1 of Fgf10 do not consistently result in functional knockout. The functional outcome depends on the specific reading frame and may be influenced by alternative translation initiation and protein domain architecture. These findings highlight important considerations for designing genome editing strategies to achieve complete gene inactivation. Full article

Sugar starvation during seed germination requires coordinated regulation of reserve mobilization, redox homeostasis, and intracellular recycling. In lupin seeds, asparagine is a major nitrogen-rich metabolite, but its role in starvation-induced autophagy and redox regulation remains unclear. Here, isolated embryonic axes of white lupin (Lupinus albus L.) and Andean lupin (Lupinus mutabilis Sweet) were cultured in vitro under sucrose-fed or sugar-starved conditions, with or without asparagine supplementation. Using transcriptomic, proteomic, immunoblot, enzymatic, antioxidant activity, and confocal microscopy analyses, we show that sugar starvation induced redox- and autophagy-related reprogramming, including changes in reactive oxygen species (ROS)-related proteins, catalase accumulation, autophagy-related (ATG) gene expression, vacuolar hydrolase-related responses, and proteolytic activity. Peroxisome-associated components, including glycolate oxidase, acyl-CoA oxidase, and catalase, were strongly affected, indicating dynamic remodeling of peroxisome-related metabolism during starvation. Asparagine modified this response by increasing antioxidant capacity and catalase accumulation under sugar starvation, while reducing detectable autophagosome number, many ATG and vacuolar hydrolase transcripts, and proteolytic activity. Together with previous evidence for asparagine-induced accumulation of autophagic bodies in vacuoles, these results are consistent with asparagine-dependent modulation of several autophagy-related processes rather than with an effect restricted to a single autophagic step. White and Andean lupin shared the same general regulatory framework but differed in response intensity. Thus, asparagine links nitrogen status with redox stabilization, vacuolar catabolism, and autophagy-related dynamics in sugar-starved lupin embryonic axes. Full article

Peptides as signaling molecules play an important role in intercellular communication. Exogenous peptides AlaGluAspLeu (AEDL) and AlaGluAsp(OMe)Leu (AED(OMe)L) at a concentration of 10⁻⁷ M stimulate the growth and development of Nicotiana tabacum. A detailed study of Nicotiana tabacum root cells grown in the presence of exogenous peptides AEDL and AED(OMe)L using transmission electron microscopy revealed characteristic differences in the ultrastructure of some cytoplasmic organelles compared to control cells. Importantly, vacuoles and autophagosomes differing in size and content, as well as amyloplasts and proteinoplasts never previously described in the literature, were found in the cells of the outer and inner root cortex. Only lytic vacuoles were detected in the cytoplasm of control cells, whereas in the presence of peptides, predominantly protein-storing vacuoles were found in root cells. In the presence of the exogenous short peptide AEDL, tobacco root cells contained amyloplasts with numerous large starch granules in the stroma, which were not detected in control cells. In the presence of the modified short peptide AED(OMe)L, leukoplasts contained protein bodies. Moreover, in contrast to control cells and cells grown in the presence of the AEDL peptide, a megaphagic (pexophagic) variant of autophagosomes with peroxisomes was detected for the first time in tobacco cells treated with AED(OMe)L. Characteristic types of phagophores were identified, forming numerous small autophagosomes with cytoplasmic regions, multivesicular bodies or concentric membranes, and cytoskeletal elements. Data on the expression of the ATG, TOR, and FREE1 genes confirmed the pattern of the existence of a large number of small autophagosomes. Based on the obtained data, a scheme for the regulation of the formation of root architecture of Nicotiana tabacum in the presence of AEDL and AED(OMe)L was proposed. Full article

Lead (Pb) pollution is a global public health issue, yet the mechanisms underlying Pb-induced kidney aging remain poorly understood. Selenium (Se), an essential trace element critical for kidney health, primarily exerts its physiological roles via selenoproteins; among them, membrane-bound selenoproteins strategically localized at the endoplasmic reticulum and plasma membrane, have emerged as potential molecular links in Se-mediated protection. In this study, Hyline chicken models (treated with 350.00 mg/L Pb or/and 1.00 mg/kg Se) and HK-2 cell models (treated with 200 μM Pb or/and 2.5 μM Se) were established to investigate the protective mechanism of Se to Pb poisoning in kidneys, with emphasis on membrane-bound selenoproteins and kidney aging. Results showed that Pb significantly decreased (p < 0.05) membrane-bound selenoproteins and induced kidney aging. Inflammation and autophagy were involved, as Pb significantly increased (p < 0.05) pro-inflammatory cytokines interleukin-4 (IL-4) and interleukin-12β (IL-12β), and autophagy-related genes autophagy related 5 (ATG5), BCL2 interacting coiled coil protein 1 (Beclin 1), and microtubule-associated protein 1A/1B-light chain 3 (LC3-II) while significantly suppressing (p < 0.05) interleukin-2 (IL-2) and mammalian target of rapamycin (mTOR). Se had a relieving effect on it, as evidenced by significantly reversing (p < 0.05) the changes in the above indicators. Interestingly, Selenoprotein T (SELENOT), as a hub membrane-bound selenoprotein, is sensitive to Pb poisoning (Pb exposure decreased SELENOT mRNA expression to 56%, 34%, and 19% of the control levels at 30, 60, and 90 days, respectively). Knockdown of SELENOT aggravated Pb-induced inflammation, autophagy, and cellular senescence, meaning that SELENOT protected against kidney aging via suppressing inflammation and autophagy under Pb stress. These findings establish membrane-bound selenoproteins as essential molecular hubs that orchestrate inflammation resolution and autophagy in Se’s antagonism against Pb-induced kidney aging, highlighting promising therapeutic targets for nephropathy. Full article

Autophagy is a crucial mechanism in the host response to intracellular bacterial pathogens during which microorganisms may undergo direct degradation in autophagolysosomes. As a highly virulent intracellular pathogen, Francisella tularensis has developed survival strategies to escape from the phagosome, replicate in the cytosol of mononuclear cells, and avoid degradation within the double-membrane vacuole during the autophagy-mediated response. The aim of this study was to investigate the role of the ATG5 autophagy protein in the host immune response to Francisella tularensis subsp. holarctica, live vaccine strain (LVS), since ATG5 plays an important role in autophagosome formation during canonical autophagy. In vitro experiments were conducted on immortalized bone marrow macrophages subjected to starvation-induced autophagy. Transgenic mice deficient in ATG5 of cells of the myeloid lineage (monocytes/macrophages and granulocytes) were used to analyze the immunological responses after intradermal infection. Cytokine levels were analyzed using Luminex, RT-qPCR, and ELISA, while inflammatory cell infiltration in the lung was analyzed by immunohistochemistry. Our results demonstrate that induced autophagy decreased bacterial replication in vitro. However, ATG5 deficiency in myeloid cells in vivo significantly diminished levels of pro-inflammatory cytokine IFN-γ in the sera, spleen, liver, and lung during Francisella infection. The attenuated pro-inflammatory response also led to significantly reduced macrophage and T cell infiltration in the lung tissue. Our findings also reveal that neutralization of IL-1β in myeloid ATG5ΔMye mice increased susceptibility to tularemia by increasing bacterial burden in organs. Full article

Objective: The present study aimed to investigate the neuroprotective effects of agomelatine in a rat model of cerebral ischemia/reperfusion injury, a major pathological event underlying ischemic stroke. Methods: Male Sprague Dawley rats included in the study were divided into four groups (n = 10/group): sham, CI/R, CI/R + 20 mg/kg Agm and CI/R + 40 mg/kg Agm. Sixty minutes of ischemia was induced in all groups except the sham group. One hour after ischemia, hydroxyethyl cellulose was administered intraperitoneally to the CI/R group, while 20 and 40 mg/kg agomelatine was administered to CI/R + Agm groups. During the three-day reperfusion period, the rats underwent neurological deficit score (NDS), rotarod, adhesive removal, and grip strength tests. At the end of the experiment, animals were decapitated and brain tissues were collected. In the collected brain tissues, infarct area was determined by TTC staining, and levels of apoptosis (Bcl-2, Bax) and autophagy (Beclin-1, ATG5, ATG7, p62) proteins were determined by Western blot. Statistical analysis of the obtained data was performed. Results: Compared with the CI/R group, agomelatine-treated groups showed significantly lower NDSs and adhesive removal times, as well as higher rotarod retention times and grip strength values (p < 0.05). Infarct area was significantly reduced following agomelatine treatment (p < 0.05). Agomelatine increased Bcl-2, Beclin-1, ATG5, and ATG7 protein levels while decreasing Bax and p62 expression compared with the CI/R group (p < 0.05). Conclusions: Agomelatine attenuated neurological deficits and infarct formation following CI/R injury. These neuroprotective effects may be associated with suppression of apoptosis and enhancement of autophagy-related pathways. Full article

Goose astrovirus (GoAstV) is a newly identified virus affecting geese, leading to goose gout, which is marked by urate deposits in organs and joints. This research investigates the role of autophagy in renal damage caused by GoAstV in goslings. The findings indicated that GoAstV infection in goslings resulted in characteristic clinical manifestations, with renal tissues displaying tubular swelling, inflammatory infiltration, and autophagosome formation. In vivo experiments demonstrated a significant upregulation of mRNA levels for autophagy-related factors, including AMPK, LC3A, ATG5, ATG7, P62, Beclin1, AMBRA1 and GABARAPL1, while mTOR and LC3B levels were notably decreased. At 3 dpi, the protein expression levels of ATG5, Beclin1, and LC3B II/I increased, while P62 levels decreased, suggesting autophagy activation. In vitro analyses revealed that GoAstV infection led to enhanced autophagy; however, the concurrent upregulation of LC3B II/I and P62 proteins suggested an obstruction in the autophagic flux. Upon the inhibition of autophagy with 3-methyladenine (3-MA, autophagy inhibitor), there was a significant reduction in the expression of autophagy-related factors, accompanied by a marked decrease in viral replication rates. In conclusion, GoAstV infection in gosling kidney cells initially triggers enhanced autophagy and subsequently causes a blockage in the autophagic flux. The virus exploits autophagosomes for replication, ultimately resulting in kidney damage. The application of 3-MA effectively inhibits this autophagic process and diminishes viral replication. Full article