Structural biology has played a key role in revealing the molecular basis of life in the past 50 years. Thanks to several landmark technical advances, the field has made significant strides from determining static macromolecular structures at atomic resolution to observing functional dynamics of protein structures spanning a vast conformational space and beyond the crystal lattice. As innovations in structural biology continue to transform modern biology and medicine, this chapter presents a perspective on past triumphs, current challenges, and future directions with a focus on how to capture protein structure dynamics using experimental and analytical methods.

Porcine rotavirus (PoRV) is a significant pathogen that causes diarrhea in piglets, with the G9, G5, and G4 genotypes being the most prevalent in China. Although vaccination is the most effective strategy to prevent PoRV infections, the currently available G5 genotype-based vaccine offers limited cross-protection against other circulating PoRV genotypes in pig farms. In this study, we developed an entirely plasmid-based reverse genetics (RG) system for the PoRV strain NJ2012 (G9P[7]) and generated two recombinant reporter viruses expressing the fluorescent UnaG and NLuc proteins, respectively. Furthermore, we successfully constructed multivalent recombinant PoRV strains by inserting the VP7 gene of G4 genotype into the gene segment 7 (NSP3) and/or the VP7 gene of G5 genotype into the gene segment 5 (NSP1) within the backbone of rNJ2012-WT strain. These multivalent recombinant viruses efficiently expressed G4 and/or G5 genotype of the VP7 protein in infected cells and elicited robust immune responses in mice. The adult mice immunized with the trivalent recombinant PoRV (rNJ2012-fG5-VP7/haG4-VP7), which simultaneously expressed VP7 proteins from G4, G5, and G9 genotypes, conferred passive protection to suckling mice against infections caused by multiple G genotypes of PoRV. In summary, these findings established a platform for efficient generation of multivalent recombinant PoRV, offering a scalable methodology to facilitate the development of next-generation PoRV vaccines.IMPORTANCEPorcine rotavirus (PoRV) is a primary etiological agent of diarrhea in swine, posing significant challenges due to the diversity of circulating genotypes and the limited cross-protection offered by existing PoRV vaccines. To address this, we developed multivalent recombinant vaccine candidates capable of eliciting robust immunity against multiple genotypes of PoRV strains. Using the established entirely plasmid-based reverse genetics (RG) system based on the G9 genotype of PoRV strain NJ2012 (G9P[7]), we further engineered the multivalent recombinant virus simultaneously expressing VP7 proteins from G4, G5, and G9 genotypes. Immunization of adult mice with this trivalent recombinant virus conferred broad-spectrum passive protection to their suckling mice against infections by multiple PoRV genotypes. Our findings established a novel platform for efficiently developing multivalent PoRV vaccines, offering a promising strategy for the prevention and control of PoRV outbreaks.

BACKGROUND/OBJECTIVES:Low-fishmeal diets are widely adopted to improve sustainability in shrimp aquaculture, yet reduced palatability and metabolic stress frequently suppress feed intake and growth. We evaluated whether a crayfish (Procambarus clarkii) by-product protein hydrolysate (CBPH) could mitigate low-fishmeal-induced performance losses by modulating feeding-related metabolic signaling and gut microbiota features in Pacific white shrimp (Litopenaeus vannamei). METHODS:In an 8-week feeding trial, 360 juveniles (initial body weight 0.46 g) were assigned to three diets (four replicates per diet): a commercial control (CON), a low-fishmeal diet (LFM), and LFM supplemented with 2% CBPH (CBPH). Growth, feed utilization, whole-body composition, hemolymph biochemical indices (TP, TG, GLU, AST, ALT), intestinal appetite-related gene expression (5-HTR, CART, CCK1R, D2-like, NPY), and intestinal microbiota profiles (full-length 16S rRNA sequencing, V1-V9, PacBio) were assessed. RESULTS:Compared with the LFM group, CBPH supplementation increased feed intake and improved feed conversion, restoring final body weight and growth rates to levels comparable to CON. CBPH also alleviated low-fishmeal-associated metabolic stress, including reduced AST and ALT activities and lower glucose levels. The LFM diet induced upregulation of anorexigenic genes (5-HTR, CART, D2-like) and downregulation of NPY in the shrimp intestine, whereas CBPH supplementation reversed these transcriptional changes. In addition, microbiota richness indices (ACE and Chao1) were elevated by CBPH relative to LFM, accompanied by compositional shifts at the phylum and genus levels. CONCLUSIONS:CBPH effectively alleviated low-fishmeal-induced reductions in feeding and growth, accompanied by coordinated changes in feeding-related gene expression, systemic biochemical markers, and gut microbiota composition, supporting its potential as a functional ingredient to stabilize metabolic responses in low-fishmeal shrimp feeds.

Prolonged exposure to aflatoxin B1 (AFB1) poses a significant threat to livestock production. The liver is the main target, but the role of the gut-liver axis and lipid metabolism in pig hepatic toxicity is not well understood. This study evaluates the impact of AFB1 on piglet liver injury via the gut-liver axis using multiomics analysis. Chronic AFB1 exposure significantly impaired the piglet growth and induced liver injury. Meanwhile, AFB1 caused gut microbiota dysbiosis and intestinal barrier damage in the piglets. Fecal microbiota transplantation (FMT) demonstrated that AFB1-altered microbiota causally contribute to hepatic inflammation in mice. Multiomics analysis revealed systemic disruption of lipid metabolism pathways, which might be involved in the intestinal flora imbalance caused by AFB1. Abnormal lipid metabolism subsequently leads to the accumulation of inflammatory lipid mediators in the plasma, ultimately causing severe liver damage. The findings highlight the crucial roles of gut microbiota and lipid metabolism in AFB1-induced liver toxicity.

Prolonged exposure to aflatoxin B1 (AFB1) poses a significant threat to livestock production. The liver is the main target, but the role of the gut-liver axis and lipid metabolism in pig hepatic toxicity is not well understood. This study evaluates the impact of AFB1 on piglet liver injury via the gut-liver axis using multiomics analysis. Chronic AFB1 exposure significantly impaired the piglet growth and induced liver injury. Meanwhile, AFB1 caused gut microbiota dysbiosis and intestinal barrier damage in the piglets. Fecal microbiota transplantation (FMT) demonstrated that AFB1-altered microbiota causally contribute to hepatic inflammation in mice. Multiomics analysis revealed systemic disruption of lipid metabolism pathways, which might be involved in the intestinal flora imbalance caused by AFB1. Abnormal lipid metabolism subsequently leads to the accumulation of inflammatory lipid mediators in the plasma, ultimately causing severe liver damage. The findings highlight the crucial roles of gut microbiota and lipid metabolism in AFB1-induced liver toxicity.

As metabolic disorders associated with excessive lipid deposition in skeletal muscle rise, strategies to alleviate this accumulation have emerged as a major focus in metabolic research. Glycerol-3-phosphate acyltransferase 3 (GPAT3) is a key enzyme in triglyceride biosynthesis; however, its role in muscle lipid metabolism remains unclear. In this study, we overexpressed or knocked down GPAT3 in C2C12 myoblasts to evaluate its effects on muscle lipid metabolism and then validated the findings in GPAT3 knockout (KO) mice. GPAT3 deficiency significantly reduced lipid deposition in muscle cells. In C2C12 cells, changes in GPAT3 expression were accompanied by alterations in AMPK signaling activity. These results demonstrate that GPAT3 promotes lipid deposition in skeletal muscle. Our in vitro data reveal a correlation between GPAT3 and AMPK signaling; further in vivo validation and functional assays are required to clarify the role of AMPK in this process. This study highlights GPAT3 as a potential therapeutic target for metabolic diseases characterized by excessive intramuscular lipids.

Protein palmitoylation, the only reversible lipid-linked post-translational modification, acts as a critical regulatory mechanism for modulating protein function and subcellular localization. However, its specific roles and underlying mechanisms in the inflammatory response of liver macrophages (Kupffer cells) remain largely undefined. This study aimed to elucidate the precise mechanism by which palmitoylation regulates inflammation in Kupffer cells and to explore potential therapeutic interventions targeting this modification. Through metabolomic and membrane proteomic analyses, we demonstrated that LPS induces metabolic reprogramming, disrupts lipid homeostasis, and elicits palmitic acid accumulation in Kupffer cells. This lipid overload promotes NLRP3 palmitoylation at conserved cysteines Cys126 and Cys898, which in turn facilitates its translocation to the trans-Golgi network membrane and subsequent inflammasome activation. Targeting this palmitoylation switch thus represents a promising therapeutic strategy for inflammatory liver diseases. We further identified the natural compound celastrol as an effective inhibitor of NLRP3 palmitoylation. Mechanistically, this effect may be cooperatively mediated by the regulation of intracellular lipid metabolism and the covalent binding of celastrol to NLRP3. Our results uncover a novel mechanistic link between metabolic dysregulation and inflammasome activation in Kupffer cells mediated by palmitoylation. Importantly, we highlight celastrol as a promising therapeutic agent that targets immunometabolic crosstalk in the pathogenesis of inflammatory liver diseases.

Uric acid to high-density lipoprotein cholesterol ratio (UHR) is a novel index of metabolism and inflammation proposed by recent studies. The predictive value of UHR has been validated in non‑alcoholic fatty liver disease. However, the association of UHR is undetermined in patients with hypertension. The aim of this study was to investigate the association of UHR with all-cause mortality and CVD mortality in patients with hypertension. This study was conducted in a real-world setting using data from a large-scale, population-based registry in China. The outcome was all-cause and CVD mortality. Cox proportional hazards models were used to investigate the associations between UHR and mortality from all-causes and cardiovascular. Penalized spline method, the Kaplan-Meier method, and subgroup analyses were also used. 13719 patients with hypertension were included in the study. During a median follow-up of 4.3 years,772 all-cause mortality occurred, with 378 CVD mortality. The mean baseline UHR was 12.53 ± 5.34. Overall, higher UHR was conspicuously associated with an elevated risk of CVD mortality and all cause mortality. Accordingly, compared with participants in quartiles 1, a conspicuous higher risk of all-cause mortality (HR = 1.52, 95

Kupffer cells (KCs), the predominant resident macrophages in the liver, exhibit an inflammatory activation state that is pathologically linked to various hepatic disorders. Studies have shown that macrophages undergo metabolic reprogramming under inflammatory conditions, and the expressions of glucose and lipid metabolism-related factors change significantly. However, glycerol kinase (GK), as a related factor that links glycolipid metabolism, the role of GK in inflammatory conditions, and its mechanism have not been reported. The aim of the present study was to explore the role of GK in the inflammatory response of KCs. LPS challenge induced marked dysregulation of glucose and lipid metabolic profiles, accompanied by a significant elevation in GK expression in pro-inflammatory KCs. GK significantly decreased the expression of pro-inflammatory factors in LPS-treated KCs. Further studies found that GK can alleviate the level of LPS-stimulated reactive oxygen species (ROS) and the expression of antioxidant factors. Meanwhile, the results showed that GK alleviates LPS-induced KCs inflammation through inhibiting the p38/STAT3 signaling pathway. The results of this study are the first to reveal that GK may alleviate Kupffer cells’ inflammatory responses by inhibiting the p38/STAT3 signaling pathway and mitigating LPS-induced ROS generation. The findings provide a potential reference for future development of drugs targeting GK to prevent KCs inflammation and even liver damage.

Photosynthesis in the world's oceans is primarily conducted by phytoplankton, microorganisms that use many different pigments for light capture. Synechococcus is a unicellular cyanobacterium estimated to be the second most abundant marine phototroph, with a global population of 7 × 1026 cells. This group's success is partly due to the pigment diversity in their photosynthetic light harvesting antennae, which maximize photon capture for photosynthesis. Many Synechococcus isolates adjust their antennae composition in response to shifts in the blue:green ratio of ambient light. This response was named type 4 chromatic acclimation (CA4). Research has made significant progress in understanding CA4 across scales, from its global ecological importance to its molecular mechanisms. Two forms of CA4 exist, each correlated with the occurrence of one of two distinct but related genomic islands. Several genes in these islands are differentially transcribed by the ambient blue:green light ratio. The encoded proteins control the addition of different pigments to the antennae proteins in blue versus green light, altering their absorption characteristics to maximize photon capture. These genes are regulated by several putative transcription factors also encoded in the genomic islands. Ecologically, CA4 is the most abundant of marine Synechococcus pigment types, occurring in over 40% of the population oceanwide. It predominates at higher latitudes and at depth, suggesting that CA4 is most beneficial under sub-saturating photosynthetic light irradiances. Future CA4 research will further clarify the ecological role of CA4 and the molecular mechanisms controlling this globally important form of phenotypic plasticity.

Cysteine is a key immunoregulatory nutrient in colorectal cancer (CRC). Cystine uptake is primarily mediated by the cystine/glutamate transporter SLC7A11, which consists of 12 transmembrane α-helices that forming a channel through the cell membrane. Given SLC7A11’s role in both metabolic competition and resistance to ferroptosis, we aim to explore whether SLC7A11 can be effectively targeted with monoclonal antibodies as a therapeutic strategy for colorectal cancer. Using a cell-based in vivo immunoassay and standard hybridoma technology, we generated a human SLC7A11-specific murine monoclonal antibody, designated 1A4. Subcutaneous and orthotopic colorectal cancer models in mice, as well as an AOM/DSS-induced colitis-associated colorectal cancer model was established. We evaluated the effects of the SLC7A11 antibody on the immune microenvironment of colorectal cancer by multicolor immunofluorescence and flow cytometry. Furthermore, single-cell sequencing was employed to analyze alterations in immune cell subpopulations and to investigate the impact of the SLC7A11 antibody on immune cell metabolism and intercellular interactions within the tumor microenvironment. Flow cytometry was further utilized to validate the differential effects of the SLC7A11 antibody on CD4+ and CD8+ T cells. Subcutaneous and orthotopic cancer model of colon cancer showed that treatment with 1A4 significantly reduced tumor volume. Multiplex immunofluorescence analysis showed that both duel immune checkpoint blockade and 1A4 increased the number of infiltrating CD8+ T cells and the CD8+T/Treg ratio in tumor tissues, with a more pronounced increase observed in tumors treated with 1A4. Single-cell sequencing results demonstrated that 1A4 modulated the tumor immune microenvironment by increasing the infiltration of T cells, B cells, and NK cells within the tumor tissue. RNA sequencing on CD4+ T cells exhibited more pronounced alterations in metabolic pathways. The flow cytometry results showed that treatment of CD8+ T cells with 1A4 enhanced stemness and a reduction in the proportion of exhausted CD8+ T cells. This study developed an antibody targeting the SLC7A11 and validated its anti-tumor effects in colorectal cancer. Targeting SLC7A11 potently creates a favorable immune microenvironment through inhibiting the activation of Tregs and increasing the stemness of CD8+ T cells. Jichang Li, Xiaoxue Pan, Kang Xia, Zeruo Yang, Xiaojing Yang, Pengyuan Wang, Shanwen Chen. Targeting SLC7A11 creates a favorable immune microenvironment in colorectal cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 2696.

Endurance exercise significantly enhances energy expenditure with lipids serving as a crucial energy source for skeletal muscle during exercise. The adipocytokine zinc-α2-glycoprotein (ZAG) in endurance exercise remains largely uncertain. This study utilized ZAG knockout and overexpression mice to investigate ZAG's role in regulating lipid metabolism in skeletal muscle during endurance exercise. Results showed the serum ZAG level of mice was significantly increased after exercise, and ZAG knockout mice decreased the exercise performance. Subsequent research revealed that ZAG knockout notably elevated triglyceride (TG) level in skeletal muscle and reduced the expression of lipolysis-related factors such as adipose triglyceride lipase (ATGL), carnitine palmitoyl transferase-1b (CPT1b), and acyl-CoA synthetase long chain family member 1, while enhancing the expression of lipid synthesis factor fatty acid synthase during exercise. The expressions of mitochondrial energy metabolism related factors uncoupling protein 2 and cytochrome c oxidase subunit I were reduced in ZAG knockout mice during endurance exercise. To assess ZAG's impact on lipid metabolism in skeletal muscle, we used ZAG overexpression plasmid in mice and C2C12 cells. ZAG overexpression decreased TG levels, enhanced ATGL expression, and increased CPT1b expression. In conclusion, ZAG can improve the level of skeletal muscle lipid metabolism and mitochondrial function during exercise, and improve the endurance exercise performance of mice.

Schizochytrium sp., a marine alga prized for docosahexaenoic acid (DHA), was subjected to UV mutagenesis to boost industrial yields. The stable mutant UV1-3 achieved 5.01 g/L DHA-40.34% higher than wild-type S31. Transcriptomic and metabolomic analyses demonstrated that UV1-3 promotes docosahexaenoic acid (DHA) biosynthesis through coordinated metabolic regulation. Downregulation of key fatty acid synthase (FAS) pathway genes (ACSL, SLC27A2, FabI) reduced substrate competition for DHA precursors. Concurrently, RT-qPCR confirmed the upregulation of core polyketide synthase (PKS) pathway genes (orfA, orfB, orfC), directly enhancing DHA production. Furthermore, suppressed oxidative phosphorylation (evidenced by COX downregulation) and redirected carbon/nitrogen flux-achieved through diminished tricarboxylic acid (TCA) cycle activity (via downregulation of HAL and proC)-collectively favored DHA accumulation. These findings establish UV1-3 as a high-yielding industrial strain for DHA production and provide fundamental insights into metabolic flux regulation in Schizochytrium sp. These insights advance scalable, cost-effective microbial DHA production and deepen understanding of algal biosynthesis mechanisms, supporting sustainable omega-3 sourcing strategies.

Inflammatory bowel disease (IBD) is a chronic inflammatory bowel disease with unclear causes and limited treatment options. Sodium butyrate (NaB), a byproduct of dietary fiber in the intestine, has demonstrated efficacy in treating inflammation. However, the precise anti-inflammatory mechanisms of NaB in colon inflammation remain largely unexplored. This study aims to investigate the effects of NaB on dextran sulfate sodium (DSS)-induced colitis in rats. The findings indicate that oral administration of NaB effectively prevent colitis and reduce levels of serum or colon inflammatory factors. Additionally, NaB demonstrated in vitro inhibition of RAW264.7 inflammation cytokines induced by LPS, along with suppression of the ERK and NF-κB signaling pathway activation. Moreover, NaB mitigated LPS and Nigericin-induced RAW264.7 pyroptosis by reducing indicators of mitochondrial damage, including increased mitochondrial membrane potential (JC-1) levels and decreased Mito-ROS production. NaB increases ZO-1 and Occludin expression in CaCo2 cells by inhibiting RAW264.7 pyroptosis. These results suggest that NaB could be utilized as a therapeutic agent or dietary supplement to alleviate colitis.

Cyanobacteriochromes are photoreceptors that constitute a significant subset of phycocyanobilin-bound proteins, yet the details of their excited-state photochemical and structural dynamics have not been fully elucidated. Here, we investigate the photoisomerization dynamics of a newly identified far-red/orange light-absorbing cyanobacteriochrome using femtosecond-resolved fluorescence and absorption methods. We observed active-site relaxations ranging from a few to hundreds of picoseconds for both far-red and orange-absorbing states. As such relaxations modulate the potential energy landscape of the chromophore, we also observed a unique dynamic spectral tuning in the far-red-absorbing state and an apparent dynamic Stokes shift in the orange-absorbing state in the femtosecond-resolved fluorescence spectra. We found that the isomerization reactions in both states occur within 320-400 ps. The observed correlation of the local relaxation and the phycocyanobilin twisting can be critical to the subsequent conformational changes after isomerization through the conical intersection to reach the final biological functions. Understanding of the time scales of the local relaxations and isomerization reactions is important to guide the design and engineering of phycocyanobilin-based light-sensitive systems of desired optical properties via synthetic biology.

Previous research has identified bile acids (BAs) as a valuable supplement for animal feed, especially in the poultry industry. However, there is limited research on the use of bile acids as a preventative measure against intestinal inflammation in broilers. This study aims to investigate the impact of dietary BAs on LPS-triggered intestinal inflammation in broilers. 180 Arbor Acres broilers were randomly divided into four group: (1) broilers receiving a standard diet (Con group); (2) broilers from the Con category subjected to LPS challenge (LPS group); (3) broilers on a diet supplemented with BAs compound and exposed to LPS (BA+LPS group); and (4) broilers on a diet enriched with lithocholic acid (LCA) and challenged with LPS (LCA + LPS group).The results showed that the LPS challenge caused a notable rise in liver mass, plasma AST concentrations, and levels of inflammatory cytokines (P < 0.05). BAs compounds or LCA improved intestinal morphological damage, inflammation response and bile acid metabolism (P < 0.05). Furthermore, analysis of 16S rRNA gene sequences revealed that supplementation with BAs compounds or LCA mitigated the reduction in bacterial diversity, while also increasing the abundance of operational taxonomic units (OTUs) associated with Bacteroides and Bifidobacterium. Additionally, the increased abundance of Candidatus_Arthromitus due to BAs compound or LCA supplementation showed a significant negative correlation with the concentrations of intestinal inflammatory cytokines (P < 0.05). These results suggest that the supplementation of BAs compound or LCA has the potential to alleviate intestinal inflammation and regulate gut microbiota in broilers subjected to LPS challenge.

This study investigated dietary regimens on Hu sheep meat quality. Compared to control, supplementary feeding (SF) increased body weight (63.83 %), eye muscle area (89.72 %), and dry matter content (5.99 %) (P < 0.01), while restricted feeding (RF) impaired meat color and reduced intramuscular fat (38.89 %) (P < 0.01). SF elevated concentrations of 67 volatile flavor compounds versus control and 48 compounds versus RF, enhancing sweet- and fatty-flavor associated volatiles. Control and RF exhibited similar flavor profiles. Correlation analysis identified 90 significant associations between fatty acids/amino acids and key volatile compounds. RF upregulated lipolytic enzymes ATGL and CPT1A (P < 0.05), decreased muscle fatty acid content, reduced muscle fiber size (8.85 %), and increased fiber density (113.50 %) (P < 0.01). Conversely, SF activated the AMPK-mTOR-S6K1 pathway, enhancing protein synthesis and amino acid levels. These findings demonstrate that dietary interventions modulate flavor precursors through metabolic pathways, providing scientific support for developing high-quality mutton products.

The inflammatory response is crucial in the progression of various liver diseases. Many diseases are linked with inflammation, such as ischemia and reperfusion injury, nonalcoholic steatohepatitis (NASH) and hepatic fibrosis. Research indicates that αKG attenuates NASH. However, the protective effect of αKG on KCs cell inflammation and its mechanism remain unclear. This study aimed to explore the role and mechanism of αKG in LPS-induced inflammation. The inflammatory damage model of KCs was induced by LPS. The study found that αKG supplementation significantly decreased inflammatory and antioxidant indexes in the LPS group, with a notable reduction in LPS-induced inflammation-related factors (P < 0.05). iNOS and COX2 are also used as indicators of macrophage inflammation, and αKG significantly alleviates LPS induced iNOS and COX2 levels (P < 0.05). Further detection of mitochondrial function and oxidative stress indexes showed that αKG supplementation significantly restored mitochondrial damage caused by LPS, inhibited ROS production, restored mitochondrial function, and improved cellular antioxidant capacity (P < 0.05). Finally, it was found that LPS treatment significantly promoted the activation of PKCε/MAPK/P65 signaling pathway, and αKG supplementation significantly inhibited the activation of signaling pathway and alleviated the pro-inflammatory effect of LPS (P < 0.05). The study demonstrated that αKG enhances the anti-inflammatory effects of KCs by decreasing ROS production and inhibiting the PKCε/MAPK/P65 signaling pathway. This is helpful for development the αKG related anti-inflammation drugs to relieving the liver inflammatory response.

The development of nonalcoholic fatty liver disease (NAFLD) may worsen due to chronic stress or prolonged use of glucocorticoids. Glycerol-3-phosphate acyltransferase 3 (GPAT3), has a function in obesity and serves as a key rate-limiting enzyme that regulates triglyceride synthesis. However, the precise impact of GPAT3 on corticosterone (CORT)-induced NAFLD and its underlying molecular mechanism remain unclear. For our in vivo experiments, we utilized male and female mice that were GPAT3-/- and wild type (WT) and treated them with CORT for a duration of 4 weeks. In our in vitro experiments, we transfected AML12 cells with GPAT3 siRNA and subsequently treated them with CORT. Under CORT-treated conditions, the absence of GPAT3 greatly improved obesity and hepatic steatosis while enhancing the expression of genes involved in fatty acid oxidation, as evidenced by our findings. In addition, the deletion of GPAT3 significantly inhibited the production of reactive oxygen species (ROS), increased the expression of antioxidant genes, and recovered the mitochondrial membrane potential in AML12 cells treated with CORT. In terms of mechanism, the absence of GPAT3 encouraged the activation of the glycogen synthase kinase 38 (GSK38)/nuclear factor-erythroid 2 related factor 2 (Nrf2) pathway, which served as a defense mechanism against liver fat accumulation and oxidative stress. Furthermore, GPAT3 expression was directly controlled at the transcriptional level by the glucocorticoid receptor (GR). Collectively, our findings suggest that GPAT3 deletion significantly alleviated hepatic steatosis and oxidative stress through promoting GSK38/Nrf2 signaling pathways.