The African swine fever virus (ASFV) has been reported to cause oxidative damage and inhibit IFN-β production. However, the precise mechanisms through which ASFV-induced lipid peroxidation modulates the innate immune response remain to be elucidated. Here, we investigated the regulatory mechanism underlying ASFV A151R-induced lipid peroxidation and ferroptosis in restricting IFN-β production The results demonstrated that A151R negatively inhibited IFN-β production via the cGAS-STING pathway by facilitating ROS accumulation, lipid peroxidation, and the carbonylative modification of STING. Concurrently, the translocation of STING from the ER to the Golgi, the generation of GSH, and the system xc−/GSH/GPX4 axis were impaired. However, GPX4 activation ameliorated lipid peroxidation mediated by A151R. Importantly, A151R facilitated NCOA4 mediated ferritinophagy. The downregulation of NCOA4 suppressed ferroptosis and lipid peroxidation, upregulated GPX4 expression, and attenuated ferroptosis. GPX4 activation abolished A151R-induced protein carbonylation, subsequently activating the TBK1-IRF3 pathway and enhancing the transcription of IFN-β and ISGs. Consistent with the in vitro findings, ASFV infection significantly reduced GPX4 and FTH levels in porcine lungs and spleens. Pharmacological inhibition of ferroptosis and knockdown of A151R in ASFV enhanced the transcription of INF-β and ISGs, reduced lipid peroxidation, and restored the expression of GPX4. Overall, our study reveals a novel mechanism whereby ASFV A151R induces STING carbonylation and triggers ferritinophagy-induced ferroptosis, thereby impairing cGAS-STING-mediated antiviral immunity. This work establishes a new paradigm for understanding ferritinophagy-driven ferroptosis and provides mechanistic insights into ASFV immune evasion strategies.

Renal Cell Carcinoma (RCC), composed of various subtypes, faces a prognostic prediction challenge. Ferroptosis, an emerging cell death form, has been considered as a vital factor for the development of cancer. However, it remains a further exploration of the predictive value for ferroptosis-correlated genes in RCC. In our work, we paid attention to the gene expression data related to clinical profiles of RCC patients which derived from public databases, TCGA and ICGC. The least absolute shrinkage and selection operator (LASSO) Cox regression performed to establish an optimally predictive model based on the TCGA data. The RCC patients derived from ICGC applied to the validation of the predictive model above. Under expectation, our observation indicated that major ferroptosis-correlated genes (75%) expressed significantly between tumorous and adjacently normal cells in the TCGA cohort. Combined with the differentially expressed genes (DEGs) (P< 0.05) and overall survival (OS), there were twenty-seven genes eventually approached the subsequent analyses. With the optimal 12-genes model screened by LASSO analysis, we aimed to classify RCC patients into the high-risk or low-risk groups based on the median scores. And we created a novel predictive model based on the 12-genes’ risk score. Compared with the two groups, our work discovered that patients in the high-risk group presented more observably shrunken OS than the low (P < 0.05). Through the multivariate cox regression, the predictive model became an independent risk factor in OS (HR =2.053, P<0.001). Moreover, the receiver operating characteristic (ROC) curve proved the significantly predictive value for our predictive model. Functional research unexpectedly showed the immune cells and immune function pathways enriched differently, which emphasized the immune role for the progress of RCC. In a word, an emerging ferroptosis-correlated signature might be popular for medical prediction in RCC. The targeted treatment on ferroptosis may be a potential schedule for RCC.

To perform a bibliometric visualization in lipidomics-related research with two decades. The primary data was retrieved from the Web of Science, three sotwares (VOSviewer, CiteSpace, and R) provided an overview of this field. The countries, institutions, authors, key terms, and keywords were tracked and corresponding mapping was generated. From January 1st in 2001 to March 21th in 2022, 45,325 authors from 234 organizations in 101 countries published 7,338 publications in 382 journals were found. Journal of Lipid Research was the most productive (284 publications) and highly cited journal (18,293 citations). We clustered four keywords themes. The niche theme were shotgun lipidomics, tandem mass-spectrometry, and electrospray-ionization. The motor theme were expression, diseases, and inflammation. The emerging or decling theme were identification, mass-spectrometry, and fatty acids.The basic theme were metabolism, cell, and plasma. Though eight categories the lipid were classified, the keywords showed two of which were got more attention for research, fatty acyls and glycerophospholipids. The top 3 lipidomics-favoured diseases were insulin resistance, obesity, and Alzheimer’s disease. The top 3 lipidomics-favoured tissue was plasma, brain, and adipose tissue. Burst citations show “women” and “pregnancy” with the strength of 8.91 and 7.1, both topics may be a potential hotspot in the future.

Peroxisome proliferator-activated receptor gamma (PPARγ) is a master regulator of adipogenesis and lipogenesis. To understand its roles in fiber formation and fat deposition in skeletal muscle, we successfully generated muscle-specific overexpression of PPARγ in two pig models by random insertion and CRISPR/Cas9 transgenic cloning procedures. The content of intramuscular fat was significantly increased in PPARγ pigs while had no changes on lean meat ratio. PPARγ could promote adipocyte differentiation by activating adipocyte differentiating regulators such as FABP4 and CCAAT/enhancer-binding protein (C/EBP), along with enhanced expression of LPL, FABP4, and PLIN1 to proceed fat deposition. Proteomics analyses demonstrated that oxidative metabolism of fatty acids and respiratory chain were activated in PPARγ pigs, thus, gathered more Ca2+ in PPARγ pigs. Raising of Ca2+ could result in increased phosphorylation of CAMKII and p38 MAPK in PPARγ pigs, which can stimulate MEF2 and PGC1α to affect fiber type and oxidative capacity. These results support that skeletal muscle-specific overexpression of PPARγ can promote oxidative fiber formation and intramuscular fat deposition in pigs.

PPARγ is a master regulator of adipogenesis and lipogenesis. To understand its roles in fat deposition and fiber formation in skeletal muscle, we successfully generated muscle-specific overexpression of PPARγ by pig cloning procedures. The content of intramuscular fat was significantly increased in PPARγ transgenic pigs while the muscle mass as lean percentage of body weight was not changed. The proteomics analysis demonstrated oxidative metabolism of fatty acids and respiratory chain were increased in PPARγ pigs. Furthermore, expressions of oxidative muscle fiber-related genes such as MyHC1 and TNNT1 were enhanced. CAMK2, MEF2 and PGC1α were also significantly increased in skeletal muscle of PPARγ pigs, indicating that Ca 2+ -sensitive phosphatases and kinases may play an important role in the switch of muscle fiber types when PPARγ activity is elevated in skeletal muscle. The results support skeletal muscle-specific overexpression of PPARγ can promote oxidative fiber formation and intramuscular fat deposition in pigs.

Individual skeletal muscles in the animal body are heterogeneous, as each is comprised of different fiber types. Type I muscle fibers are rich with mitochondria, and have high oxidative metabolisms while type IIB fibers have few mitochondria and high glycolytic metabolic capacity. Peroxisome proliferator-activated receptor gamma coactivator 1α (PGC-1α), a transcriptional co-activator that regulates mitochondrial biogenesis and respiratory function, is implicated in muscle fiber-type switching. Over-expression of PGC-1α in transgenic mice increased the proportion of red/oxidative type I fiber. During pig muscle growth, an increased number of type I fibers can give meat more red color. To explore the roles of PGC-1α in regulation of muscle fiber type conversion, we generated skeletal muscle-specific PGC-1α transgenic mice and pig. Ectopic over-expression of PGC-1α was detected in both fast and slow muscle fibers. The transgenic animals displayed a remarkable amount of red/oxidative muscle fibers in major skeletal muscle tissues. Skeletal muscles from transgenic mice and pigs have increased expression levels of oxidative fiber markers such as MHC1, MHC2x, myoglobin and Tnni1, and decreased expressions of glycolytic fiber genes (MHC2a, MHC2b, CASQ-1 and Tnni2). The genes responsible for the TCA cycle and oxidative phosphorylation, cytochrome coxidase 2 and 4, and citrate synthase were also increased in the transgenic mice and pigs. These results suggested that transgenic over-expressed PGC-1α significantly increased muscle mitochondrial biogenesis, resulting in qualitative changes from glycolytic to oxidative energy generation. The transgenic animals also had elevated levels of PDK4 and PPARγ proteins in muscle tissue, which can lead to increased glycogen deposition and fatty acid oxidation. Therefore, the results support a significant role of PGC-1α in conversion of fast glycolytic fibers to slow and oxidative fiber through enhanced mitochondrial respiration and fatty acid oxidation, and transgenic over-expression of PGC-1α in skeletal muscle leads to more red meat production in pigs.

It is a general consensus that oocyte quality is the key to embryo survival in pig reproduction. Thus, study on regulation of the ovary-associated gene is of great significance in pig breeding. Peptidylarginine deiminases (PADs) are a family of enzymes which catalyze the conversion of arginine to citrulline in proteins. The peptidylarginine deiminases type VI gene (PADI6) is mainly expressed in the ovary, and plays an important role in oocyte growth, fertilization and early embryo development. However, until now, little is known about its transcriptional regulation mechanism. Here, we firstly isolated and characterized the 5′-flanking region of porcine PADI6 gene. We determined the transcription start site using 5′-rapid amplification of cDNA ends (RACE) analysis, and identified the minimal promoter (− 85/+68) that drove the basal expression of PADI6 by constructing various progressive deletions. Mutational analysis and electrophoretic mobility shift assays demonstrated Sp1 bound to the − 56/− 47 region of the PADI6 promoter. Furthermore, overexpression of Sp1 significantly increased the promoter activity and promoted PADI6 gene expression, and accordingly, inhibition of Sp1 expression with specific siRNA significantly reduced the promoter activity and suppressed the PADI6 expression. In addition, inhibition of Sp1 binding by Mithramycin A treatment reduced the transcriptional activity of PADI6 in a dose-dependent manner. Taken together, these data indicate that Sp1 is essential for the transcriptional regulation of PADI6.

Visfatin is an adipocytokine displaying multiple functional properties, which plays a role in the regulation of cell apoptosis and inflammation by an as yet unidentified mechanism. The aim of the present study was to determine if visfatin is involved in apoptosis pathway induced by LPS in rat Mesenteric lymph nodes (MLNs). Experimental rats were divided into four groups and MLNs samples were collected from each group. The morphological changes of the MLNs were examined by histological imaging. CD68 and ENPP1 were detected with immunohistochemistry and Western Blot. Apoptosis was evaluated with TUNEL and Flow Cytometry, the mRNA levels of the apoptosis-related genes were detected by qRT-PCR, and the protein levels of the apoptotic-related factors were detected by western blot. The main results showed that visfatin could significantly increase the macrophages in MLNs and prevent cell apoptosis from LPS-induced mesenteric lymph nodes, activate apoptotic signaling pathways and regulate the mRNA levels of the apoptosis-related genes. Visfatin had a pro-apoptotic effect on normal MLNs, whereas it exerted an anti-apoptotic effect during LPS-induced cell apoptosis in rat MLNs. In short, visfatin plays a dual role in the apoptosis in rat MLNs, which is mediated by both the mitochondrial apoptotic pathway and the death-receptor apoptotic pathway.

FAM3A (family with sequence similarity 3, member A) is regulated by PPARG and participates in the metabolism of lipid in liver. However, the transcriptional regulation analysis of FAM3A is very little and biological function of FAM3A still unclear. In this study, the core promoter region and transcription factor binding sites of FAM3A gene were identified and characterized using dual luciferase report experiments and electrophoretic mobility shift assays (EMSA). The promoter activity of FAM3A was dramatically decreased after the mutation of C/EBPβ binding sites, suggesting that C/EBPβ is a transcriptional activator of FAM3A. Overexpression of FAM3A significantly inhibited the efficiency of preadipocytes to differentiate into adipocytes as indicated by Western Blot and Oil Red O staining assay. These results suggest that C/EBPβ plays an important role in regulating FAM3A promoter activity and FAM3A inhibits adipocyte differentiation.

This study was undertaken to determine if visfatin is involved in the inflammation or apoptosis introduced by LPS in rats. Forty 8-week old Wistar rats were divided into four groups (n=10 in each group) and injected with saline, visfatin, LPS and visfatin+LPS co-stimulated via caudal vein. The duodenum, jejunum and ileum were harvested from all the rats. Compared to the saline treated group, visfatin significantly increased the number of TUNEL-positive apoptotic cells and the expression of caspase-3 protein in intestinal mucosa. Similarly, ELISA and western blot analysis also showed the up-regulation of pro-caspase-3 and cleaved caspase-3 expression in the visfatin group compared to the control group. In contrast to LPS, visfatin down-regulated the expression of cleaved-caspase-3 in the visfatin+LPS co-stimulated group, resulting in a significant decrease in apoptosis in intestinal mucosal cells. We observed more pro-caspase-3 positive cells in the visfatin+LPS co-stimulated group. The results indicate that, in the presence of LPS, visfatin plays an important role in the regulation of cell apoptosis and inflammation.

The purpose of the present study is to determine if visfatin is involved in inflammation or apoptosis induced by LPS in rat. Forty Wistar rats were divided into four groups: saline group, LPS group, visfatin group and Visfatin + LPS co-stimulated group. Spleen samples from each group of rats were collected for study. The spleen structure was examined by histological imaging. Apoptosis was evaluated with TUNEL reaction. Caspase-3 was detected with immunohistochemistry and western blot. The apoptosis-related genes were detected by qPCR and inflammatory cytokines were tested by ELISA. Our main findings were as follows. (1) Macrophages were markedly increased in the visfatin group compared with the saline group. This finding was confirmed when spleen samples were examined with western blot using CD68 antibody. (2) Visfatin promoted the expression of CD68 and caspase-3 in rat spleen, whereas visfatin could inhibit the expression of CD68 and activated caspase-3 in spleen of LPS-induced acute inflammation. (3) Visfatin had a pro-apoptotic effect on normal rat spleen, whereas it exerted an anti-apoptotic effect during LPS-induced lymphocytes apoptosis in rat spleen. Moreover, the effect of visfatin on cell apoptosis was mediated by the mitochondrial pathway. (4) Visfatin could modulate both the anti-inflammatory cytokines and pro-inflammatory cytokines in rat spleen, such as IL-10, IL-4, IL-6, TNF-α and IL-1β. Taken together, we demonstrate that visfatin could participate in the inflammatory process in rat spleen by modulating the macrophages and inflammatory cytokines. Also, visfatin plays a dual role in the apoptosis in rat spleen, which is mediated by the mitochondrial pathway.

Este estudio investigo los efectos de la visfatina sobre la estructura y los niveles de inmunidad en el intestino delgado de ratas inducidas por lipopolisacaridos (LPS). Cuarenta ratas Wistar se dividieron aleatoriamente e igualmente en cuatro grupos: solucion salina (control), vistafin, LPS y visfatina + LPS co-estimuladas. Las funciones de la visfatina en la inmunidad de la mucosa intestinal se investigaron mediante el examen de variacion de la estructura del tejido, la inflamacion y las proteinas relacionadas con la inmunidad en el intestino de ratas estresadas inmunologicamente; usando tincion HE, ELISA, inmunohistoquimica y Western Blot. Los resultados mostraron que, en comparacion con el grupo control, el grupo tratado con visfatina presento una disminucion en la altura y ancho de las vellosidades intestinales, y un aumento significativo en los niveles de IL-6 y TNF-ð, asi como inmunoglobulina A (IgA celulas positivas). Ademas, al comparar este grupo con el grupo tratado con LPS- el grupo visfatina + LPS co-estimulado mostro una disminucion en la altura y ancho de las vellosidades, asi como en los niveles de IL-6 y TNF-ð, y un aumento en los niveles de IgA, lo que implica reduccion de una respuesta a la inyeccion LPS. Todos los resultados sugieren que, en condiciones fisiologicas normales, la visfatina perturba la homeostasis del cuerpo y provoca la atrofia de las vellosidades intestinales mediante el aumento de la expresion de IgA. Mientras que bajo condiciones de la respuesta inmune, LPS actua como un antigeno exogeno para promover visfatina contra la inflamacion inducida por LPS por la disminucion de la expresion de IgA. En condiciones de estres inmunologico, la visfatina como estimulo exogeno promueve la respuesta inmune mediante la regulacion de los niveles de proteina de IL-6, TNF-ð e IgA.

Brown adipose tissue (BAT) is specialized to dissipate energy as heat, therefore reducing fat deposition and counteracting obesity. Brown adipocytes arise from myoblastic progenitors during embryonic development by the action of transcription regulator PRDM16 binding to PPARγ, which promotes BAT-like phenotype in white adipose tissue. To investigate the capability of converting white adipose tissue to BAT or browning by PPARγ in vivo, we generated transgenic mice with over-expressed PPARγ2. The transgenic mice showed strong brown fat features in subcutaneous fat in morphology and histology. To provide molecular evidences on browning characteristics of the adipose tissue, we employed quantitative real-time PCR to determine BAT-specific gene expressions. The transgenic mice had remarkably elevated mRNA level of UCP1, Elovl3, PGC1α and Cebpα in subcutaneous fat. Compared with wild-type mice, UCP1 protein levels were increased significantly in transgenic mice. ATP concentration was slightly decreased in the subcutaneous fat of transgenic mice. Western blotting analysis also confirmed that phosphorylated AMPK and ACC proteins were significantly (P<0.01) increased in the transgenic mice. Therefore, this study demonstrated that over-expression of PPARγ2 in skeletal muscle can promote conversion of subcutaneous fat to brown fat formation, which can have beneficial effects on increasing energy metabolisms and combating obesity.

Low cloning efficiency is considered to be caused by the incomplete or aberrant epigenetic reprogramming of differentiated donor cells in somatic cell nuclear transfer (SCNT) embryos. Oxamflatin, a novel class of histone deacetylase inhibitor (HDACi), has been found to improve the in vitro and full-term developmental potential of SCNT embryos. In the present study, we studied the effects of oxamflatin treatment on in vitro porcine SCNT embryos. Our results indicated that the rate of in vitro blastocyst formation of SCNT embryos treated with 1 μM oxamflatin for 15 h postactivation was significantly higher than all other treatments. Treatment of oxamflatin decreased the relative histone deacetylase (HDAC) activity in cloned embryos and resulted in hyperacetylation levels of histone H3 at lysine 9 (AcH3K9) and histone H4 at lysine 5 (AcH4K5) at pronuclear, two-cell, and four-cell stages partly through downregulating HDAC1. The suppression of HDAC6 through oxamflatin increased the nonhistone acetylation level of α-tubulin during the mitotic cell cycle of early SCNT embryos. In addition, we demonstrated that oxamflatin downregulated DNA methyltransferase 1 (DNMT1) expression and global DNA methylation level (5-methylcytosine) in two-cell-stage porcine SCNT embryos. The pluripotency-related gene POU5F1 was found to be upregulated in the oxamflatin-treated group with a decreased DNA methylation tendency in its promoter regions. Treatment of oxamflatin did not change the locus-specific DNA methylation levels of Sus scrofa heterochromatic satellite DNA sequences at the blastocyst stage. Meanwhile, our findings suggest that treatment with HDACi may contribute to maintaining the stable status of cytoskeleton-associated elements, such as acetylated α-tubulin, which may be the crucial determinants of donor nuclear reprogramming in early SCNT embryos. In summary, oxamflatin treatment improves the developmental potential of porcine SCNT embryos in vitro.