BACKGROUND:Recent advances in swine gut culturomics have substantially expanded our understanding of the cultivable repertoire and revealed the compositional complexity of the porcine gut microbiota. Nevertheless, how multiple environmental and cultivation-related factors interact to shape cultured communities under controlled nutrient conditions remains poorly understood. This study evaluated the relative and interactive contributions of intestinal segment, oxygen availability, cultivation mode, and carbon-substrate identity within a standardized nutrient-rich basal medium framework. Using inocula from the ileum and colon of Wuzhishan piglets, a roughage-tolerant Chinese indigenous breed, microbial communities were cultured on modified yeast extract casitone fatty acid (YCFA) media in which the original carbon substrate was replaced with ten alternative carbohydrates representing simple and complex substrates. RESULTS:Culture-dependent (CD) sequencing showed that intestinal segment was the dominant factor shaping cultured communities (P < 0 0.001), followed by oxygen availability (P < 0 0.001) and donor identity (P < 0 0.001), whereas, when evaluated within a nutrient-rich basal medium, carbon-substrate identity did not function as an independent or strongly additive driver of community differentiation (P > 0 0.05). A total of 1,308 isolates belonging to 67 genera were obtained through an empirical streaking strategy, including several rare taxa such as Culturomica that were below sequencing detection thresholds, revealing a disparity between CD sequencing profiles and actual strain isolation. Integration of community composition with short-chain fatty acid (SCFA) profiles further indicated that metabolic accumulation represents an additional ecological dimension influencing cultured community differentiation. CONCLUSIONS:Our findings demonstrate that under multi-factor cultivation frameworks, ecological filters do not operate in a freely additive manner. Instead, intestinal origin, oxygen exposure, and metabolic context collectively constrain culturability, while carbon-substrate variation alone does not independently drive community clustering within nutrient-rich conditions. Together, this study provides a refined conceptual framework for interpreting culture-dependent outcomes and for designing multi-factor cultivation strategies in future swine gut culturomics research.

This study investigated the impact of dietary tryptophan levels on carcass traits, meat quality, flavor profiles and antioxidant capacity in finishing pigs under large-scale farming conditions. Approximately 400 finishing pigs were randomly allocated to four groups, each with four replicate pens. The pigs received diets containing either the recommended level of tryptophan or an increased level of tryptophan by 15%, 35%, or 55%, for a period of 31 days. The results showed that dietary tryptophan had no significant influence on growth performance or carcass traits. While meeting the tryptophan requirements, increasing dietary tryptophan levels by 35% reduced the b* value and shear force of pork. Moreover, a 35% increase in tryptophan levels weakened the fruity, green, fatty and fresh flavor profiles, as indicated by decreased levels of key volatile flavor compounds (VOCs). These included the fruity-associated 6-methyl-5-hepten-2-one and (E,E)-3,5-octadien-2-one, the fresh and fruity-related hexanoic acid methyl ester and ethyl 2-ethylhexanoate, as well as the green and fatty odor-contributing nonanal. Notably, increasing dietary tryptophan levels increased antioxidant capacity and tryptophan metabolite content, particularly nicotinamide, which were associated with changes in key VOCs. This study deepens our understanding of how dietary tryptophan levels affect pork flavor and provides valuable insights for improving pork quality through nutritional regulation.

The rice blast fungal effector AVR-PikC binds to the rice protein HIPP19, which may contribute to plant susceptibility. The compound B93 induces the interaction between the rice E3 ligase APIP6 and AVR-PikC, which results in the ubiquitination and degradation of AVR-PikC, thereby facilitating plant resistance.

African swine fever virus (ASFV) employs sophisticated regulatory strategies to manipulate host cell apoptosis, a process critical for its pathogenesis and immune evasion; however, the mechanisms underlying this process remain incompletely understood. Here, we report a novel mechanism by which the ASFV-encoded envelope protein CD2v suppresses apoptosis by activating the TPL2 (tumor progression locus 2)-MEK (mitogen-activated protein kinase kinase)-ERK (extracellular signal-regulated kinase) signaling axis, leading to proteasomal degradation of the pro-apoptotic protein BimEL in primary porcine alveolar macrophages and wild boar lung (WSL) cells. We further demonstrated that ASFV infection triggers ERK1/2-dependent phosphorylation and degradation of BimEL, a process independent of viral replication and mediated by viral structural components. A targeted screen identified CD2v as the key viral protein driving this pathway. Both the purified extracellular domain of CD2v (Asp17-Tyr206) and virion-associated CD2v activated TPL2-MEK-ERK signaling without requiring internalization into cells, resulting in BimEL downregulation and subsequent suppression of apoptosis. Crucially, CRISPR-Cas9-mediated knockout of CD2v abolished ASFV-induced ERK1/2 activation and consequential BimEL degradation. Furthermore, we discovered that soluble CD2v released from ASFV-infected cells can activate this signaling axis in uninfected bystander cells, thereby inhibiting apoptosis distantly. This paracrine function, alongside its intrinsic role in directly infected cells, enables CD2v to establish a pro-survival microenvironment conducive to viral propagation. Our findings uncover a multifaceted anti-apoptotic mechanism employed by ASFV, expanding the functional repertoire of CD2v and providing new insights into ASFV pathogenesis with potential therapeutic implications.IMPORTANCEThis study elucidates a distinct mechanism of apoptosis inhibition by African swine fever virus (ASFV), a pathogen that causes a devastating disease in swine. We identify the ASFV CD2v protein as a key suppressor of cell death that operates by hijacking the host TPL2-MEK-ERK signaling pathway to degrade the pro-apoptotic protein BimEL. Importantly, CD2v mediates this effect not only within infected cells but also, in a soluble form, on surrounding uninfected bystander cells. This dual action helps create a protective, pro-survival cellular environment that facilitates viral spread and persistence. Understanding this novel apoptotic suppression mechanism advances our knowledge of ASFV-host interactions and highlights potential new avenues for therapeutic intervention.

Supplementation with Lactobacillus strains attracts intense interest for its potential in regulating fat accumulation in both humans and animals. However, the functional disparities among Lactobacillus strains remain poorly understood. This study examined the impact of dietary supplementation with Lactobacillus reuteri (L. reuteri) and Lactobacillus johnsonii (L. johnsonii) on lipid metabolism, carcass traits, and meat quality in heavy finishing pigs. A total of 288 finishing pigs (equal numbers of castrated barrows and gilts; initial body weight = 109.88 ± 1.58 kg) were randomly allocated to 3 dietary treatments with 6 replicate pens and 16 pigs per pen. Pigs were fed a basal diet (CON), or the basal diet supplemented with L. reuteri or L. johnsonii at a daily dose of 5 × 1010 CFU per pig, respectively, over a 42-d experimental period. The experimental results demonstrated that compared with CON, both probiotic strains significantly decreased serum levels of cortisol and pro-inflammatory factors including TNF-α, IL-1β and IL-17, while significantly elevating the levels of anti-inflammatory factor IL-4 and total antioxidant capacity (T-AOC) (P < 0.05), and reduced the concentration of saturated fatty acids in Longissimus thoracis (LT) (P = 0.014). L. johnsonii supplementation significantly reduced backfat thickness at the 10th-rib (P = 0.040) and last-rib backfat (P = 0.025), significantly increased fat-free lean index (P = 0.015), and significantly decreased adipocyte size in subcutaneous adipose tissue (P < 0.001), accompanied by the activation of the AMPK/PPARγ signaling pathway. Compared with CON, L. reuteri supplementation significantly reduced the shear force (P = 0.043) and intramuscular fat (IMF) content (P = 0.026) of LT muscle of finishing pigs and significantly increased free valine, methionine, and isoleucine concentrations in fresh meat (P < 0.05). In conclusion, the two strains of Lactobacillus exhibited distinct characteristics. Specifically, L. johnsonii primarily promoted lipid catabolism while simultaneously suppressing lipid anabolism by activating the AMPK/PPARγ signaling pathway, leading to a reduction in backfat thickness. In contrast, L. reuteri exhibited stronger focus on improving meat quality traits, such as tenderness and amino acid composition. This study offers a valuable insight into the specific application of probiotics in high-quality pork production.

Inflammatory bowel disease (IBD) poses a grave threat to human health, and minipigs are physiologically more analogous to humans than rodents. Probiotics are widely recognized for their ability to regulate the immune microenvironment and improve barrier function; however, the role of Lactobacillus combination in IBD has not been fully studied. The present study was conducted to investigate the anti-inflammatory effects of Lactobacillus combination (Lactobacillus crispatus, Lactobacillus johnsonii and Lactobacillus reuteri) on dextran sodium sulfate (DSS)-treated Wuzhishan minipigs. A total of thirteen healthy Wuzhishan minipigs were randomly divided into three groups: a CON group, a DSS group, and a DSS + LAB group. On days 8 and 23 of the experiment, the Lactobacillus mixture or DSS solution was administered to the piglets. The results demonstrated that the Lactobacillus mixture exhibited a positive effect on weight reduction and colon histopathology scores. The administration of DSS resulted in a substantial augmentation in the expression of proinflammatory cytokines, including interleukin-1 beta (IL-1β) and tumor necrosis factor alpha (TNF-α). The Lactobacillus mixture supplement was found to suppress the upregulation of proinflammatory cytokines (IL-1β) and the downregulation of anti-inflammatory cytokines (transforming growth factor beta 1 (TGF-β1)), while increasing the expression levels of tight junction protein zonula occludens-1 (ZO-1) and mucin 1 (MUC1). Furthermore, improvements in the content of oxidative stress indicators and short-chain fatty acids (SCFAs) were observed under the mediation of the Lactobacillus mixture. 16S rRNA sequencing revealed that the Lactobacillus mixture was effective in reducing the relative abundance of harmful bacteria and increasing that of beneficial bacteria, thereby reshaping the intestinal microbiota structure of Wuzhishan minipigs with DSS-induced colitis and further altering the metabolic pathways of the intestinal microbiota. This study provides a novel reference and scientific basis for the application of Lactobacillus in the treatment of colitis and for the food industry to develop functional probiotic products.

Summary statement β‐caryophyllene (BCP) plays diverse roles in plants, particularly in resistance induction and plant‐plant communication. This study reports the underlying mechanism: BCP competes with NINJA and IAA1/IAA10 for TOPLESS binding, thereby relieving the repression of JA and auxin signalling pathways.

In the North China Plain (NCP), excessive irrigation has been widely used to maintain high wheat and maize yields; however, this practice has severely depleted groundwater resources and accelerated nitrous oxide (N2O) emissions. Thus, a 3-year field experiment in a wheat-maize cropping system was conducted to explore the impacts of three different drip irrigation strategies (irrigating to 80%, 75% and 70% of the field water holding capacity [WHC] when the soil water content was less than 65% [D1], 60% [D2] and 55% [D3] of the WHC, respectively) on soil N2O emissions. Compared with D1, D2 maintained high annual grain yields while significantly reducing annual cumulative N2O emissions by 8.8%–15.7% and yield-scaled emissions by 9.1%–11.8%. In contrast, D3 reduced annual grain yields by 9.2%–15.8% compared with D1. D2 consistently stabilized soil water-filled pore space (WFPS) within 45%–50%, which lowered the average abundances of denitrification-related genes (nirS, nirK, norB and nosZ clade I) by 17.4%–32.3% compared to D1. Enzymatic activities (N-acetyl-β-D-glucosaminidase [NAG], leucine aminopeptidase [LAP], nitrate reductase [NR]) and the abundances of functional genes associated with nitrification and denitrification (amoA-AOB, nirK, nosZ clade I) were optimally regulated under D2. Multivariate analysis and structural equation modeling (SEM) revealed that N2O emissions were significantly and positively affected by soil WFPS, dissolved organic C/N, and NAG activity, with reduced irrigation water amount directly suppressing denitrification pathways to mitigate N2O emissions. Our findings highlight that irrigating to 75% WHC (D2) is an efficient strategy to balance high yield and N2O mitigation, providing a practical pathway for sustainable intensification in water-limited regions.

Straw incorporation (SI) is a widely adopted management strategy that improves soil fertility, alters soil-microbe-plant interactions, and enhances crop yield in agroecosystems. A deep understanding of soil-microbe-plant interactions in response to SI is crucial for optimizing fertilization practices and boosting crop yield; however, relevant knowledge remains limited. A two-year consecutive field experiment was conducted during winter wheat growing seasons to investigate the effects of fertilization at the level of 0 kg N ha–1 (N0) and 210 kg N ha–1 (N1) under straw incorporated (S1) and removal (S0) on soil environment, microbial community structure and plant growth and yield. SI had a significant effect on soil organic matter (SOM) content, while fertilization had a significant effect on NO3– concentration, SOM and TN contents. N1S1 decreased the root length and total surface area (TSA), but increased aboveground biomass and yield by 32.7

To address water scarcity, environmental pollution from excessive fertilization, and the need for stable grain yields in the North China Plain (NCP), a 3-year field experiment (2021-2024) was conducted to explore the effects of water and nitrogen (N) management under subsurface drip irrigation on winter wheat-summer maize rotation system. This study included three irrigation treatments (irrigated to 80% (D1), 75% (D2), and 70% (D3) of field water-holding capacity (WHC) when soil water content dropped below 65%, 60%, and 55% of WHC, respectively) under N210 and four N application rates (0 (N0), 150 (N150), 210 (N210), and 270 (N270) kg N ha-1 for each season) under D1. The results showed that D1 and N210 all significantly improved leaf area index (LAI) and aboveground biomass by reducing chlorophyllase and pheophytinase activities (delaying leaf senescence) and enhancing nitrate reductase (NR), glutamine synthetase (GS), and glutamate synthase (GOGAT) activities (promoting N assimilation). Furthermore, compared with D3, D1 increased winter wheat and summer maize yields by 12.7% and 24.3%, respectively, and improved partial factor productivity (PFP) by 14.0-46.0%. Redundancy analysis (RDA) and structural equation modeling confirmed that LAI, biomass, and N-transforming enzyme activities were the key drivers of yield. This study demonstrated that the combination of D1 and N210 treatments were the optimal water and N fertilizer management strategies for achieving water conservation, N reduction, and stable high yields in drip-irrigated rotation systems of the NCP.

Embryo loss during early pregnancy is a major constraint on mammalian reproductive efficiency. We previously revealed that dietary L-malic acid (L-MA) supplementation benefits maternal health in sows during late pregnancy. However, the effect of L-MA on sows during early pregnancy is largely unexplored. In the present study, in vitro assays confirmed that L-MA directly promoted the adhesion of embryonic trophoblast (JAR) cells to endometrial epithelial (Ishikawa) cells, and alleviated H₂O₂-induced reactive oxygen species (ROS) accumulation in the endometrial cells. In vivo, L-MA supply during early pregnancy optimized the physiological environment in sows, thereby improving pregnancy outcomes, without altering circulating estradiol or progesterone levels on GD 28. L-MA significantly enhanced maternal antioxidant capacity and alleviated inflammatory responses. Metabolomics analysis showed that L-MA reshaped the maternal metabolic profile, modulated amino acid, lipid and nucleotide metabolism, and activated the NOD-like receptor signaling pathway and the cGMP-PKG signaling pathway. L-MA enriched beneficial bacteria, including short-chain fatty acid (SCFA)-producing, anti-inflammatory, and antioxidant taxa, while repressing inflammation-associated bacteria, and these specific taxa were significantly correlated with differential metabolites, antioxidant/anti-inflammatory markers, and reproductive outcomes. Overall, our data revealed that dietary L-MA supplementation during early pregnancy enhanced reproductive performance in sows. These benefits might be associated with shifts in the maternal gut microbiota-metabolite axis and enhanced antioxidant and anti-inflammatory capacities. Therefore, L-MA supplementation may be a potential strategy for ameliorating embryo loss and improving reproductive performance in mammals.

Rice is a staple crop feeding over half of the global population, yet it faces severe yield losses due to devastating diseases, including those caused by hemibiotrophic pathogens, such as Magnaporthe oryzae (the causative agent of blast disease) and Xanthomonas oryzae pv. oryzae (the bacterial leaf blight pathogen). While resistance genes are a cornerstone of crop protection, many nucleotide-binding leucine-rich receptor ( NLR )-type resistance genes are prone to breakdown and often impose yield penalties. In this study, we report that the cross-species transfer of the maize NAM-ATAF1/2-CUC2 transcription factor ZmNAC2 into rice confers resistance to both blast and bacterial leaf blight diseases without compromising yield. Mechanistically, ZmNAC2 interacts with OsNAC2, a negative regulator of salicylic acid (SA) biosynthesis, and disrupts its association with the APETALA2/ethylene-responsive element binding protein OsEREBP1 in the OsZmNAC2 transgenic rice, thereby quenching repression and promoting SA production. Moreover, ZmNAC2 binds to the cis -regulatory elements within the promoter of the SA biosynthetic gene phenylalanine ammonia lyase 6 , transactivating its expression and further enhancing SA accumulation. The resulting elevated SA levels impart broad-spectrum resistance in the transgenic rice against M. oryzae and X. oryzae pv. oryzae . Together, our findings provide a proof of concept for leveraging non- NLR genes from staple food crops to boost disease resistance without incurring yield penalties.

As an important economic crop, tomato is vulnerable to various diseases, and these diseases often have high visual similarity, making identification difficult. A delay in diagnosis can have a significant effect on tomato yields. Traditional manual visual inspection methods have poor accuracy, while laboratory diagnostic methods are inefficient, making them unsuitable for large-scale agricultural scenarios. To address this challenge, this study involved the collection and construction of a tomato leaf disease dataset in a real planting environment, and AutoAugment was used to achieve sample diversity and balance the number of training samples in different categories. Additionally, a dual-path ensemble network (DPEN) was proposed, which combines the multiscale feature extraction advantages of GoogLeNet with the dense connection mechanism of DenseNet121. The experimental results show that, compared with the comparison models, the DPEN achieves an identification precision of 98.80% on the self-built dataset, which is an improvement of 2.33% to 9.24%, and a reduction in the number of parameters by 7.09 M compared with GoogLeNet and 2.07 M compared with DenseNet121. The experimental results on public datasets further demonstrated the accuracy of the proposed DPEN model in identifying tomato leaf diseases in complex backgrounds. These results prove that the DPEN model can achieve precise, rapid, and efficient identification of tomato leaf diseases in complex backgrounds, providing technical support for smart agriculture applications.

Tumor necrosis factor receptor-associated factors (TRAFs), as key signal transducers, play a crucial role in both innate and adaptive immune responses. However, current research on the systematic identification of the TRAF family and its functional role in Siberian sturgeon (Acipenser baerii) is relatively unexplored. In this study, TRAF family members were first identified in A. baerii using comparative genomics and transcriptome analyses. To further determine the TRAF sequences of A. baerii, the open reading frame (ORF) sequences of the Abtraf1, Abtraf5, Abtraf6, and Abtraf7 were cloned. Bioinformatics analysis revealed that these Abtraf exhibit high sequence similarity and evolutionary conservation across teleost species. RNA-seq and qRT-PCR analysis revealed high Abtraf expression in the whole pre-larvae stage and immune-related tissues such as the head kidney, gill, and spleen. Induction of Abtraf gene expression was observed upon stimulation with Streptococcus iniae and Aeromonas hydrophila in vivo. In this study, head kidney leukocytes (HKLs) were isolated and stimulated with lipopolysaccharide (LPS), peptidoglycan (PGN), or poly(I:C), all of which induced Abtraf expression, with significant upregulation occurring early post-treatment. Notably, Abtraf5 responded predominantly to bacterial components (LPS/PGN), whereas Abtraf7 was specifically poly(I:C)-responsive. These findings help clarify the molecular roles of Abtraf and provide a basis for further functional research on TRAF in teleosts.

Rice production is threatened by blast disease caused by Magnaporthe oryzae. Application of fungicides remain one of the main controlling strategies. Due to the emergence of resistant strains, developing fungicides with novel mechanisms has become necessary. The Pmk1 kinase of M. oryzae is essential for fungal virulence, particularly in appressoria formation and invasive hyphae expansion within the host. Herein, we demonstrate that Pmk1 can serve as a potential target for fungicide development, and present the critical sites of Pmk1 for inhibitor design. Through high-throughput screening, we obtained the compound A59, which interacts with Pmk1 and inhibits its activity. In vivo assays showed that A59 suppresses appressoria development and invasive hyphae expansion of M. oryzae in the hosts, manifesting both protective and therapeutic effects. With an integration of computational and experimental validation, we demonstrated that A59 binds to the catalytic pocket of Pmk1 and identified the critical sites of Pmk1 for inhibitor interaction. These findings were further supported by the evaluation of chiral isomers of A59. This study presents Pmk1 as a novel fungal MAPK target for developing fungicides with protective and therapeutic effects, and identifies Pmk1 critical residues for inhibitor interactions, providing a foundation for rational design of Pmk1-targeted fungicides.

Cortisol can impact the transcription of downstream inflammation and immune-related genes via the glucocorticoid receptor (GR), thereby influencing the immune response and maintaining the homeostatic balance of the host. However, there is a lack of research on the mechanisms by which cortisol affects the immune response of Siberian sturgeon (Acipenser baerii) through GR. In this study, an anti-inflammatory state of Siberian sturgeon was established by the combined treatment of head kidney (HK) leukocytes with LPS + cortisol. Subsequently, the inflammation-related genes of the AbGR antagonistic group (LPS + cortisol + RU-486) and the AbGR non-antagonistic group (LPS + cortisol) were compared by qRT-PCR and high-throughput sequencing methods. Furthermore, an AP-1 agonist ASLAN003 was used to detect the regulatory effect of the AP-1 gene on inflammatory cytokines. The results showed that cortisol downregulated the expressions of il-1β, il-6, il-8, tnf-α, and il-17c that were induced by LPS, while simultaneously promoting the expressions of tgf-β1. Moreover, this pattern was reversed by adding RU486. When analyzing the differentially expressed genes in the transcriptome sequencing of HK leukocytes in AbGR antagonistic group, 261 significantly down-regulated genes and 194 significantly up-regulated genes were annotated. Furthermore, 26 differentially expressed genes related to inflammation in AbGR antagonistic group were enriched, and the key nuclear transcription factor AP-1 for regulating inflammation function of AbGR was selected based on the enrichment factor and p-value for subsequent research. In the HK leukocytes after cortisol + ASLAN003 treatment, the expressions of fosl1 and jund that were induced by ASLAN003 were significantly down-regulated after cortisol treatment. In this situation, the expression of tgf-β1 was significantly increased, and the expression of tnf-α was significantly decreased after cortisol treatment. Therefore, this study demonstrated that cortisol inhibits the expression of AP-1 through GR in Siberian sturgeon, and then regulates the generation of the inflammatory response.

Gastrulation marks a pivotal stage in mammalian embryonic development, establishing the three germ layers and body axis through lineage diversification and morphogenetic movements. However, studying human gastrulating embryos is challenging due to limited access to early tissues. Here we show the use of spatial transcriptomics to analyse a fully intact Carnegie stage 7 human embryo at single-cell resolution, along with immunofluorescence validations in a second embryo. Employing 82 serial cryosections and Stereo-seq technology, we reconstructed a three-dimensional model of the embryo. Our findings reveal early specification of distinct mesoderm subtypes and the presence of the anterior visceral endoderm. Notably, primordial germ cells were located in the connecting stalk, and haematopoietic stem cell-independent haematopoiesis was observed in the yolk sac. This study advances our understanding of human gastrulation and provides a valuable dataset for future research in early human development.

SGT1 (the suppressor of the G2 allele of Skp1) functions as an adaptor protein that positively regulates plant defense and developmental processes. It comprises three functional domains: the tetratricopeptide repeat (TPR) domain, Chord SGT1 motif (CS), and SGT1-specific motif (SGS). In this study, we resolved the crystal structure of the Oryza sativa OsSGT1-TPR domain at 1.53 Å resolution. Structural analysis showed that the TPR domain adopts a homo-dimeric architecture stabilized by salt bridges (mediated by K52/R79/R109) and hydrophobic interactions (involving F17). Functional validation through gel filtration chromatography revealed that the disruption of the dimerization interface via F17A/K52A/R79A mutations caused complete dissociation into monomers, establishing the essential role of TPR-mediated oligomerization in maintaining the structural stability of full-length OsSGT1. Yeast two-hybrid assays showed that the dimerization disruption of SGT1 mutants retained the interaction with OsHSP81-2 (an HSP90 ortholog) and OsRAR1, indicating that SGT1 oligomerization serves primarily as a structural stabilizer rather than a prerequisite for partner interaction. Evolutionary analysis through the sequence alignment of plant SGT1 proteins revealed the conservation of the dimerization interface residues. This study provides structural insights into the conserved molecular features of SGT1 proteins and highlights the functional significance of their oligomerization state.

The various environmental pressures could increase the cortisol levels of sturgeon. However, there is a lack of reports on the mechanism by which elevated cortisol levels affect the immune response of Siberian sturgeon. In this study, a high-level cortisol anti-inflammatory state of Siberian sturgeon after co-treatment with LPS and cortisol was constructed and verified in the primary spleen leukocytes. The glucocorticoid receptor (GR) was cloned and its binding site with Ru486 was analyzed. After adding Ru486 to treat the primary spleen leukocytes, the expression of inflammatory cytokines mediated by GR was explored by qRT-PCR. The structure of spleen after LPS + cortisol injection was observed by histopathology and the changes in apoptotic genes were explored by qRT-PCR. Furthermore, the apoptosis of cells after Ru486 treatment was studied by qRT-PCR, DNA ladder, and Hoechst staining. Firstly, co-treatment with LPS and cortisol led to high levels of serum cortisol and glucose within 24 h after treatment. In this situation, the expression of il-10 and tgf-beta 1 was upregulated in the LPS + cortisol group, while the expression of il-1 beta and tnf-alpha was regulated. However, il-1 beta, tnf-alpha, il-10, and tgf-beta 1 were significantly down-regulated in vitro. Clone analysis of the GR sequence indicated that it is conserved and widely expressed in 18 tissues. Molecular docking simulations suggested that it can bind to cortisol and the GR antagonist Ru486, respectively. Further addition of Ru486 treatment reversed the expression of il-1 beta, il-10, tgf-beta 1, and JAK-STAT signaling pathway factors. Histopathological observations showed that LPS + cortisol injection causes splenic cell death. Then, the expression of pro-apoptotic genes p53 and caspase3 increases significantly. After Ru486 treatment in spleen leukocytes, the mRNA expression levels of pro-apoptotic-related genes p53, bax, caspase3, caspase7, and caspase9 were significantly downregulated. GR antagonism could attenuate LPS + cortisol-treated splenic leukocyte DNA fragmentation and nuclear morphological changes. Therefore, cortisol can regulate the production of inflammatory cytokines and promote the process of cell apoptosis through the GR of Siberian sturgeon, thereby inhibiting the inflammatory response.