Oxidative Stress-related Genes Research Articles

Biological potential of Bacillus subtilis BS45 to inhibit the growth of Fusarium graminearum through oxidative damage and perturbing related protein synthesis.

Fusarium root rot (FRR) caused by Fusarium graminearum poses a threat to global food security. Biological control is a promising control strategy for FRR. In this study, antagonistic bacteria were obtained using an in-vitro dual culture bioassay with F. graminearum. Molecular identification of the bacteria based on the 16S rDNA gene and whole genome revealed that the species belonged to the genus Bacillus. We evaluated the strain BS45 for its mechanism against phytopathogenic fungi and its biocontrol potential against FRR caused by F. graminearum. A methanol extract of BS45 caused swelling of the hyphal cells and the inhibition of conidial germination. The cell membrane was damaged and the macromolecular material leaked out of cells. In addition, the mycelial reactive oxygen species level increased, mitochondrial membrane potential decreased, oxidative stress-related gene expression level increased and oxygen-scavenging enzyme activity changed. In conclusion, the methanol extract of BS45 induced hyphal cell death through oxidative damage. A transcriptome analysis showed that differentially expressed genes were significantly enriched in ribosome function and various amino acid transport pathways, and the protein contents in cells were affected by the methanol extract of BS45, indicating that it interfered with mycelial protein synthesis. In terms of biocontrol capacity, the biomass of wheat seedlings treated with the bacteria increased, and the BS45 strain significantly inhibited the incidence of FRR disease in greenhouse tests. Therefore, strain BS45 and its metabolites are promising candidates for the biological control of F. graminearum and its related root rot diseases.

Tandem mass tag-based quantitative proteomics analysis reveals the effects of the α-lactalbumin peptides GINY and DQW on lipid deposition and oxidative stress in HepG2 cells

The objective of this study was to investigate the mechanism by which the α-lactalbumin peptides Gly-Ile-Asn-Tyr (GINY) and Asp-Gln-Trp (DQW) ameliorate free fatty acid-induced lipid deposition in HepG2 cells. The results show that GINY and DQW reduced triglyceride, total cholesterol, and free fatty acid levels significantly in free fatty acid-treated HepG2 cells. Based on proteomic analysis, GINY and DQW alleviated lipid deposition and oxidative stress mainly through the peroxisome proliferator-activated receptor (PPAR) pathway, fatty acid metabolism, oxidative phosphorylation, and response to oxidative stress. In vitro experiments confirmed that GINY and DQW upregulated the mRNA and protein expression of fatty acid β-oxidation-related and oxidative stress-related genes, and downregulated the mRNA and protein expression of lipogenesis-related genes by activating peroxisome proliferator-activated receptor α (PPARα). Meanwhile, GINY and DQW reduced free fatty acid-induced lipid droplet accumulation and reactive oxygen species generation, and enhanced the mitochondrial membrane potential and ATP levels. Furthermore, GINY and DQW enhanced carnitine palmitoyl-transferase 1a (CPT-1a) and superoxide dismutase activities, and diminished acetyl-coenzyme A carboxylase 1 (ACC1) and fatty acid synthase (FASN) activities in a PPARα-dependent manner. Interestingly, GW6471 (a PPARα inhibitor) weakened the effects of GINY and DQW on the PPARα pathway. Hence, our findings suggest that GINY and DQW have the potential to alleviate nonalcoholic fatty liver disease by activating the PPARα pathway.

Expression Pattern and Molecular Mechanism of Oxidative Stress-Related Genes in Myocardial Ischemia-Reperfusion Injury.

(1) Background: The molecular mechanism of oxidative stress-related genes (OSRGs) in myocardial ischemia-reperfusion injury (MIRI) has not been fully elucidated. (2) Methods: Differential expression analysis, enrichment analysis, and PPI analysis were performed on the MIRI-related datasets GSE160516 and GSE61592 to find key pathways and hub genes. OSRGs were obtained from the Molecular Signatures Database (MSigDB). The expression pattern and time changes of them were studied on the basis of their raw expression data. Corresponding online databases were used to predict miRNAs, transcription factors (TFs), and therapeutic drugs targeting common differentially expressed OSRGs. These identified OSRGs were further verified in the external dataset GSE4105 and H9C2 cell hypoxia-reoxygenation (HR) model. (3) Results: A total of 134 DEGs of MIRI were identified which were enriched in the pathways of "immune response", "inflammatory response", "neutrophil chemotaxis", "phagosome", and "platelet activation". Six hub genes and 12 common differentially expressed OSRGs were identified. A total of 168 miRNAs, 41 TFs, and 21 therapeutic drugs were predicted targeting these OSRGs. Lastly, the expression trends of Aif1, Apoe, Arg1, Col1a1, Gpx7, and Hmox1 were confirmed in the external dataset and HR model. (4) Conclusions: Aif1, Apoe, Arg1, Col1a1, Gpx7, and Hmox1 may be involved in the oxidative stress mechanism of MIRI, and the intervention of these genes may be a potential therapeutic strategy.

Establishment and external verification of an oxidative stress-related gene signature to predict clinical outcomes and therapeutic responses of colorectal cancer.

Objective: Accumulated evidence highlights the biological significance of oxidative stress in tumorigenicity and progression of colorectal cancer (CRC). Our study aimed to establish a reliable oxidative stress-related signature to predict patients' clinical outcomes and therapeutic responses. Methods: Transcriptome profiles and clinical features of CRC patients were retrospectively analyzed from public datasets. LASSO analysis was used to construct an oxidative stress-related signature to predict overall survival, disease-free survival, disease-specific survival, and progression-free survival. Additionally, antitumor immunity, drug sensitivity, signaling pathways, and molecular subtypes were analyzed between different risk subsets through TIP, CIBERSORT, oncoPredict, etc. approaches. The genes in the signature were experimentally verified in the human colorectal mucosal cell line (FHC) along with CRC cell lines (SW-480 and HCT-116) through RT-qPCR or Western blot. Results: An oxidative stress-related signature was established, composed of ACOX1, CPT2, NAT2, NRG1, PPARGC1A, CDKN2A, CRYAB, NGFR, and UCN. The signature displayed an excellent capacity for survival prediction and was linked to worse clinicopathological features. Moreover, the signature correlated with antitumor immunity, drug sensitivity, and CRC-related pathways. Among molecular subtypes, the CSC subtype had the highest risk score. Experiments demonstrated that CDKN2A and UCN were up-regulated and ACOX1, CPT2, NAT2, NRG1, PPARGC1A, CRYAB, and NGFR were down-regulated in CRC than normal cells. In H2O2-induced CRC cells, their expression was notably altered. Conclusion: Altogether, our findings constructed an oxidative stress-related signature that can predict survival outcomes and therapeutic response in CRC patients, thus potentially assisting prognosis prediction and adjuvant therapy decisions.

Combined scRNAseq and Bulk RNAseq Analysis to Reveal the Dual Roles of Oxidative Stress-Related Genes in Acute Myeloid Leukemia.

Oxidative stress (OS) can either lead to leukemogenesis or induce tumor cell death by inflammation and immune response accompanying the process of OS through chemotherapy. However, previous studies mainly focus on the level of OS state and the salient factors leading to tumorigenesis and progression of acute myeloid leukemia (AML), and nothing has been done to distinguish the OS-related genes with different functions. First, we downloaded single-cell RNA sequencing (scRNAseq) and bulk RNA sequencing (RNAseq) data from public databases and evaluated the oxidative stress functions between leukemia cells and normal cells by the ssGSEA algorithm. Then, we used machine learning methods to screen out OS gene set A related to the occurrence and prognosis of AML and OS gene set B related to treatment in leukemia stem cells (LSCs) like population (HSC-like). Furthermore, we screened out the hub genes in the above two gene sets and used them to identify molecular subclasses and construct a model for predicting therapy response. Leukemia cells have different OS functions compared to normal cells and significant OS functional changes before and after chemotherapy. Two different clusters in gene set A were identified, which showed different biological properties and clinical relevance. The sensitive model for predicting therapy response based on gene set B demonstrated predictive accuracy by ROC and internal validation. We combined scRNAseq and bulk RNAseq data to construct two different transcriptomic profiles to reveal the different roles of OS-related genes involved in AML oncogenesis and chemotherapy resistance, which might provide important insights into the mechanism of OS-related genes in the pathogenesis and drug resistance of AML.

The Effect of N6-Methyladenosine Regulators and m6A Reader YTHDC1-Mediated N6-Methyladenosine Modification Is Involved in Oxidative Stress in Human Aortic Dissection.

Aortic dissection (AD) develops pathological changes in the separation of the true and false aortic lumen, with high lethality. m6A methylation and oxidative stress have also been shown to be involved in the onset of AD. Through bioinformatics methods, three differentially expressed m6A regulators (YTHDC1, YTHDC2, and RBM15) were excavated from the GSE52093 dataset in the Gene Expression Omnibus (GEO) database, and functional enrichment analysis of the differentially expressed genes (DEGs) regulated by m6A regulators was performed. Then, the genes with oxidative stress-related functions among these genes were found. The protein interaction network of the oxidative stress-related genes and the competing endogenous RNA- (ceRNA-) miRNA-mRNA network were constructed. Among them, DHCR24, P4HB, and PDGFRA, which have m6A differences in AD samples, were selected as key genes. We also performed immune infiltration analysis, as well as cell-gene correlation analysis, on samples from the dataset. The results showed that YTHDC1 was positively correlated with macrophage M1 and negatively correlated with macrophage M2. Finally, we extracted AD and healthy aorta RNA and protein from human tissues that were taken from AD patients and patients who received heart transplants, performed quantitative real-time PCR (qRT-PCR) on YTHDC2 and RBM15, and performed qRT-PCR and western blot (WB) detection on YTHDC1 to verify their differences in AD. The mRNA and protein levels of YTHDC1 were consistent with the results of bioinformatics analysis and were downregulated in AD. Immunofluorescence (IF) was used to colocalize YTHDC1 and endothelial cell marker CD31. After knocking down YTHDC1 in human umbilical vein endothelial cells (HUVECs), reactive oxygen species (ROS) levels had a tendency to increase and the expression of peroxide dismutase SOD2 was decreased. This study provides assistance in discovering the role of m6A regulator YTHDC1 in AD. In particular, m6A modification participates in oxidative stress and jointly affects AD.

Hydroxytyrosol's effect on the expression of apoptosis and oxidative stress related genes in BE (2)-C neuroblastoma cell line

Neuroblastoma (NB) is defined as a type of tumor that involves the autonomic nervous system. Oxidative stress (OxS) resulted from dopamine (DA) oxidation and the reduction of antioxidant enzyme activity is the main reason for the initiation and progression of NB. The aim of this study was the evaluation of the protective effects of Hydroxytyrosol (HT) against OxS induced by H2O2. In the present in vitro study, the OxS condition was induced by H2O2 treatment of BE (2)-C cells; then, the cells were treated by several concentrations of HT and Vitamin C (VC). Next, the expression levels of genes related to OxS and apoptosis were measured through the RT-qPCR method. Subsequently, the activity of catalase (CAT), glutathione peroxidase (GPx), and superoxide dismutase (SOD) was determined via enzyme colorimetric assay. Finally, the level of malondialdehyde (MDA) was measured by the Thiobarbituric acid reactive substances (TBARS) assay. HT treatment leads to a significantly decreased the expression level of Bax, caspase-3, and p53 (p < 0.05); as well, the expression levels of Nrf-2, SOD, Bcl-2, PPAR-γ, CAT, and GPx were increased at a significant manner (p < 0.05). Moreover, HT treatment led to a remarkable increase in the enzyme activity of CAT, SOD, and GPx (p < 0.001). Finally, the MDA levels were significantly reduced (p < 0.001) in BE (2)-C cells after HT treatment. The findings suggest that HT could have neuroprotective effects by enhancing various enzymatic and molecular pathways to increase the cellular antioxidant defense. Additionally, HT reduced the expression of the P53 gene and blocked the apoptotic signaling pathways by suppressing Bax and caspase-3. However, further animal models studies are needed to approve the beneficial effects of HT.

Seasonal changes in endoplasmic reticulum stress and ovarian steroidogenesis in the muskrats (Ondatra zibethicus).

Many studies have shown roles for endoplasmic reticulum stress (ERS)/unfolded protein response (UPR) signaling cascades with ovarian folliculogenesis, and oocyte maturation. In this study, we investigated seasonal changes in ERS and ovarian steroidogenesis in the muskrats (Ondatra zibethicus) during the breeding season (BS) and non-breeding season (NBS). There were noticeable seasonal variations in the weight and size of muskrat ovaries with values higher in the BS than that in NBS. The circulating luteinizing hormone (LH), follicle-stimulating hormone (FSH), 17β-estradiol, and progesterone of the female muskrats were higher during the BS. The RNA-seq data of ovaries during different seasons revealed 2580 differentially expressed genes, further analysis showed a prominent enrichment of ERS-related pathways and ovarian steroidogenesis pathway. Immunohistochemical results showed that GRP78 and steroidogenic enzymes (P450scc, 3β-HSD, P450c17, and P450arom) existed in the various kinds of cells in muskrat ovaries during the BS and NBS. In ovaries from the BS, the mRNA levels of P450scc, P450arom, P450c17, and 3β-HSD were considerably higher. Furthermore, the expression levels of oxidative stress-related genes (SOD2, CAT, and GPX1) and UPR signal genes (Bip/GRP78, ATF4, ATF6, and XBP1s) were increased strikingly higher during the BS in comparison with the NBS. However, the mRNA levels of CCAAT-enhancer-binding protein homologous protein (CHOP) and caspase-3 had no considerable difference between the BS and NBS. Taken together, these results suggested that UPR signaling associated with the seasonal changes in ovarian steroidogenesis is activated in the BS and the delicate balance in redox regulation is important for seasonal reproduction in the muskrats.

Cystic Fibrosis Transmembrane Conductance Regulator Attenuates Oxidative Stress-Induced Injury in Diabetic Retinopathy Rats

Purpose To investigate the effects of cystic fibrosis transmembrane conductance regulator (CFTR) on oxidative stress-induced injury of diabetic retinopathy (DR) rats. Methods DR rat model was constructed treated with Ad-CFTR. Hematoxylin and Eosin (HE) staining was applied for testing the thickness of each layer of retinal tissues. Enzyme-linked immunosorbent assay (ELISA) was used to determine levels of serum inflammatory cytokines and contents of oxidative stress related genes in rats. Terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL) staining was used to detect retinal cell apoptosis, and western blotting to measure the expression of MAPK/NF-κB pathway-related proteins in retinal tissues. Results Our experiment revealed the remarkable decrease of CFTR protein in retinal tissues of DR rats. DR rats had decreased body weight and increased blood glucose level, with decreased thickness of total retinal thickness (TRT), outer nuclear layer and outer plexiform layer (ONL + OPL), inner nuclear layer (INL), and inner plexiform layer (IPL). Besides, DR rats were apparently up-regulated in the expression of pro-inflammatory cytokines, with increased malondial dehyde (MDA), p-ERK1/2/ERK1/2 and p-JNK1/2/JNK1/2 expressions, decreased superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activity in retinal tissues, as well as up-regulated p65 protein in nucleus and down-regulated p65 protein in cytoplasm. DR rats treated with Ad-CFTR were effectively improved regarding the above parameters except body weight and blood glucose. Conclusions CFTR can inhibit MAPK/NF-κB signaling pathway to ameliorate inflammatory response and oxidative stress-induced injury of DR rats, thereby reducing retinal cell apoptosis and playing a protective role in retina.

Platycodin D-Induced Immunotoxicity in RAW 264.7 Macrophages via Oxidative Stress-Mediated Apoptosis

Platycodin D (PD) is a naturally occurring, biologically active triterpenoid saponin isolated from a medicinal food homology plant called Platycodon grandiflorus (Jacq.) A. DC. It is involved in the processing of various biological activities. While investigating the anti-inflammatory property of PD using lipopolysaccharide (LPS)-stimulated murine RAW 264.7 macrophage cells, we unexpectedly found that PD exhibited toxicity to RAW 264.7 cells. In this study, the toxic effect of PD on RAW 264.7 cells was systematically evaluated for the first time. The results showed that PD (12.5−200 µM) significantly reduced cell viability and inhibited cell proliferation in a dose-dependent manner. At a concentration of 20 µM, PD significantly increased lactate dehydrogenase activity and the mRNA and protein expression of Bax, p53, Casp3, IL-1β, and TNF-α. Interestingly, PD (0.8−20 µM) inhibited the expression of inflammatory cytokines in LPS-stimulated RAW 264.7 cells. PD (20 µM) also significantly increased reactive oxygen species (ROS) levels and the expression of oxidative stress-related genes and proteins. This study revealed that PD exhibited immunotoxicity to RAW 264.7 cells, with possible mechanisms including oxidative stress-mediated apoptosis resulting in activation of the mitochondrial apoptosis pathway and dysregulated expression of inflammatory cytokines. This study evaluated the impact of PD on immunity and provided guidelines for its future biological application.

A novel prognostic model related to oxidative stress for treatment prediction in lung adenocarcinoma.

The prognostic model based on oxidative stress for lung adenocarcinoma (LUAD) remains unclear. The information of LUAD patients were acquired from TCGA dataset. We also collected two external datasets from GEO for verification. Oxidative stress-related genes (ORGs) were extracted from Genecards. We performed machine learning algorithms, including Univariate Cox regression, Random Survival Forest, and Least Absolute Shrinkage and Selection Operator (Lasso) analyses on the ORGs to build the OS-score and OS-signature. We drew the Kaplan-Meier and time-dependent receiver operating characteristic curve (ROC) to evaluate the efficacy of the OS-signature in predicting the prognosis of LUAD. We used GISTIC 2.0 and maftool algorithms to explore Genomic mutation of OS-signature. To analyze characteristic of tumor infiltrating immune cells, ESTIMATE, TIMER2.0, MCPcounter and ssGSEA algorithms were applied, thus evaluating the immunotherapeutic strategies. Chemotherapeutics sensitivity analysis was based on pRRophetic package. Finally, PCR assays was also used to detect the expression values of related genes in the OS-signature in cell lines. Ten ORGs with prognostic value and the OS-signature containing three prognostic ORGs were identified. The significantly better prognosis of LUAD patients was observed in LUAD patients. The efficiency and accuracy of OS-signature in predicting prognosis for LUAD patients was confirmed by survival ROC curves and two external validation data sets. It was clearly observed that patients with high OS-scores had lower immunomodulators levels (with a few exceptions), stromal score, immune score, ESTIMATE score and infiltrating immune cell populations. On the contrary, patients with higher OS-scores were more likely to have higher tumor purity. PCR assays showed that, MRPL44 and CYCS were significantly higher expressed in LUAD cell lines, while CAT was significantly lower expressed. The novel oxidative stress-related model we identified could be used for prognosis and treatment prediction in lung adenocarcinoma.

Transcriptional profiling of extraintestinal Escherichia coli exposed to cranberry pomace extracts alone or in combination with ceftriaxone

Extraintestinal pathogenic Escherichia coli (ExPEC) includes several serotypes that have been associated with colibacillosis in poultry, as well as urinary tract infections and newborn meningitis in humans. This study investigated the antimicrobial activities of ceftriaxone (AXO) and cranberry pomace extracts (CRAN) alone or in combination (CC) against multidrug-resistant (MDR) ExPEC from broiler. The growth-inhibitory activity of CRAN and synergy tests by a checkerboard method were determined in cation-adjusted Mueller–Hinton broth (CAMHB). The transcriptomic profile of the MDR E. coli O7:H18 (ST38) grown in CAMHB supplemented with sub-inhibitory concertation of CRAN and AXO alone or in combination was obtained by RNA-seq. The MIC of CRAN for all isolates was 16 mg/mL. An additive activity was observed between 4 mg/mL of CRAN and 4 μg/mL of AXO. Compared to the control, the transcriptomic analysis revealed that 4 mg/ml of (1/4MIC) CRAN and its combination with 4 μg/mL of (1/8MIC) AXO (CC) exposures resulted in 727 and 712 differentially expressed genes, respectively (false discovery rate &amp;lt; 0.001 and log2-fold change &amp;gt; 2), in the studied E. coli. Major virulence genes including adhesins (fim, flg, csg, and yad), protectins (omp, tra, waa, and hly), secretion systems (hof, pho, and vir), and quorum sensing (lsr), which are energetically expensive for bacteria, were downregulated. Most importantly, 1/4MIC of CRAN or CC downregulated the β-lactamase blaCMY-2 and efflux pump including tolC, mdtEIJ, gadEW, and their regulator gene evgS, while upregulating the cysteine biosynthesis and oxidative stress-related regulatory genes including cys, dmlA, sbp, nrdGHI, soxSR, and rpoH. Downregulation of multiple enzymes involved in TCA cycles and upregulation of Fe–S cluster coordinated by Cys and Isc proteins reflect the regulation of energy metabolism of the studied E. coli upon CRAN or CC exposure. The downregulation of outer membrane protein genes that control permeability barriers, along with different antimicrobial resistance genes, demonstrates that CRAN may have the unique potential to enhance the antimicrobial activities of third-generation cephalosporins such as AXO against MDR E. coli.

Identification of thioredoxin-1 as a biomarker of lung cancer and evaluation of its prognostic value based on bioinformatics analysis.

Thioredoxin-1 (TXN), a redox balance factor, plays an essential role in oxidative stress and has been shown to act as a potential contributor to various cancers. This study evaluated the role of TXN in lung cancer by bioinformatics analyses. Genes differentially expressed in lung cancer and oxidative stress related genes were obtained from The Cancer Genome Atlas, Gene Expression Omnibus and GeneCards databases. Following identification of TXN as an optimal differentially expressed gene by bioinformatics, the prognostic value of TXN in lung cancer was evaluated by univariate/multivariate Cox regression and Kaplan-Meier survival analyses, with validation by receiver operation characteristic curve analysis. The association between TXN expression and lung cancer was verified by immunohistochemical analysis of the Human Protein Atlas database, as well as by western blotting and qPCR. Cell proliferation was determined by cell counting kit-8 after changing TXN expression using lentiviral transfection. Twenty differentially expressed oxidative stress genes were identified. Differential expression analysis identified five genes (CASP3, CAT, TXN, GSR, and HSPA4) and Kaplan-Meier survival analysis identified four genes (IL-6, CYCS, TXN, and BCL2) that differed significantly in lung cancer and normal lung tissue, indicating that TXN was an optimal differentially expressed gene. Multivariate Cox regression analysis showed that T stage (T3/T4), N stage (N2/N3), curative effect (progressive diseases) and high TXN expression were associated with poor survival, although high TXN expression was poorly predictive of overall survival. TXN was highly expressed in lung cancer tissues and cells. Knockdown of TXN suppressed cell proliferation, while overexpression of TXN enhanced cell proliferation. High expression of TXN plays an important role in lung cancer development and prognosis. Because it is a prospective prognostic factor, targeting TXN may have clinical benefits in the treatment of lung cancer.

Visible-Light-Driven Zinc Oxide Quantum Dots for the Management of Bacterial Fruit Blotch Disease and the Improvement of Melon Seedlings Growth.

Bacterial fruit blotch is one of the most destructing diseases of melon producing-regions. Here, zinc oxide quantum dots (ZnO QDs) were synthesized, and their antibacterial activity against Acidovorax citrulli was investigated. The results indicated that the obtained ZnO QDs displayed 5.7-fold higher antibacterial activity than a commercial Zn-based bactericide (zinc thiazole). Interestingly, the antibacterial activity of ZnO QDs irradiated with light was 1.8 times higher than that of the dark-treated group. It was because ZnO QDs could induce the generation of hydroxyl radicals and then up-regulate the expression of oxidative stress-related genes, finally leading to the loss of cell membrane integrity. A pot experiment demonstrated that foliar application of ZnO QDs significantly reduced the bacterial fruit blotch disease incidence (32.0%). Furthermore, the supply of ZnO QDs could improve the growth of infected melon seedlings by activating the antioxidant defense system. This work provides a promising light-activated quantum-bactericide for the management of pathogenic bacterial infections in melon crop protection.

The toxic effect of bisphenol AF and nanoplastic coexposure in parental and offspring generation zebrafish

Microplastics (MPs) and bisphenol AF (BPAF) are two environmental pollutants that usually coexist in the natural environment. Studies of MPs or BPAF have gradually increased in recent years, but few studies have focused on the combination toxic effects. In this study, the subchronic model of adult zebrafish was exposed to 1 mg/L nanolevel microplastics and 200 μg/L BPAF for 45 days; the parental zebrafish were spawning every 3 days during exposure, and the effects of continuous poisoning were examined on the offspring after 1–9 spawns. The results showed that single BPAF exposure or BPAF and nanoplastic coexposure can both decrease the number of eggs laid and the locomotor behavior of parental zebrafish and impact the hatching rate, mortality, body length and locomotor behavior of offspring zebrafish, especially in 7–9 spawn. BPAF were accumulated in parental zebrafish intestinal in 334.62 ng/g in BPAF group and 594.52 ng/g in nm+BPAF group, and accumulated in whole offspring zebrafish for 281.6 ng/g in BPAF group and 321.46 ng/g in nm+BPAF group. Neurodevelopmental, inflammation, apoptosis and oxidative stress-related genes were also significantly increased after 7–9 spawn. In addition, the exacerbated accumulation in the BPAF+nm group in parental and offspring zebrafish may be the reason for the accelerated toxic effect in the present research. In this study, we investigated the combined effects of nanoplastics and BPAF on parental and offspring zebrafish in the aquatic environment to identify the accumulative toxic effects and provide new experimental support for assessing the effects of coexposure on aquatic organisms.

Improving the protective ability of lignin against vascular and neurological development in BPAF-induced zebrafish by high-pressure homogenization technology

The lack of a sufficient amount of functional groups in the lignin structure limits its bioapplication. In this work, high-pressure homogenization was performed on original kraft lignin (L-ORI) to prepare lignin nanoparticles (L-NANO), which aimed to improve its functional group contents for further vascular and neurological applications. The results showed that the prepared L-NANO possessed spherical structures with diameters of 40.3–160.4 nm and increased amount of hydroxyl groups. Compared to L-ORI, L-NANO possessed better in vivo and in vitro antioxidant capacity, which could endow it with enhanced protective effects for the vascular and neural development of bisphenol AF (BPAF)-induced zebrafish. In addition, L-NANO reduced the neurotoxicity and cardiovascular toxicity of BPAF in zebrafish by upregulating the expression levels of oxidative stress-related genes (Cu/Zn-Sod and cat), which could further significantly upregulate the expression levels of neurogenesis genes (elavl3, gap43, mbp, and syn2a) and protect the contraction of the cardinal vein (CCV) and early central nervous system development by upregulating the expression levels of vascular genes (flk1 and flt4). The excellent cardiovascular and neurodevelopmental protective ability of L-NANO indicated that high-pressure homogenization is a promising technology for improving the bioactivity of lignin.

Transcriptomic profiling of lipopolysaccharide-challenged bovine mammary epithelial cells treated with forsythoside A

Mastitis is usually caused by a variety of pathogenic bacteria that seriously impact the health and milk-production ability of dairy cows, with consequent, economically detrimental effects on the dairy industry. Forsythoside A (FTA), isolated from the fruit and leaves of Forsythia suspensa (Thunb.) Vahl (Oleaceae), has been reported to have significant antioxidant, anti-inflammatory, and antibacterial effects. However, it is not clear whether FTA exerts a protective effect against lipopolysaccharide (LPS)-induced bovine mastitis and its potential gene signature. In this study, high-throughput sequencing technology was performed to analyze the differences between the mRNA and enrichment pathway of bovine mammary epithelial cells of the control, LPS, and LPS + FTA groups. The results showed that there were 139 differentially expressed genes (DEGs) (p-value < 0.05, |log2FoldChange| > 1, FPKM > 1) in the LPS group compared with the control group, including 121 up-regulated genes and 18 down-regulated genes, which were mainly enriched in the cellular response to lipopolysaccharide, cytokine activity, protein binding, and IL-17 signaling pathway based on Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, respectively. Compared with the control group and LPS + FTA group, there were 349 DEGs, including 322 up-regulated genes and 27 down-regulated genes. They were mainly enriched in protein localization to organelles, centrosomes, binding, and the IL-17 signaling pathway, based on GO and KEGG analysis. Compared to the LPS group, the LPS + FTA group had 272 DEGs, including 259 up-regulated genes and 13 down-regulated genes, which were mainly enriched in RNA processing, IL-6 receptor binding, and the lysosome pathway, based on GO and KEGG analyses. It can be seen that LPS stimulation induced the expression of inflammation-related genes, IL-17 and IL-6, whereas FTA treatment promoted the expression of the spliceosome-, lysosome-, and oxidative stress-related genes HSP70, HSPA8, and PARP2. The study utilized RNA-sequencing analysis of FTA against LPS-challenged bovine mammary epithelial cells to explore key mRNA findings that may be strongly associated with inflammation and oxidative stress, and provides a theoretical reference for further elucidation of molecular mechanisms of bovine mastitis and therapeutic effects of FTA against bovine mastitis.

A four oxidative stress gene prognostic model and integrated immunity-analysis in pancreatic adenocarcinoma.

Pancreatic adenocarcinoma (PAAD) is highly aggressive and characterized by a poor prognosis. Oxidative stress has great impacts on the occurrence and development of tumors. However, the predictive role of oxidative stress related genes on PAAD patients' prognosis remains unclear. In this study, we aimed to construct a prognostic model for PAAD based on oxidative stress genes and to evaluate its predictive value. The Cancer Genome Atlas (TCGA) and three Gene Expression Omnibus (GEO) datasets were used to identify differentially expressed oxidative stress genes. Univariate Cox regression, Kaplan-Meier and multivariate Cox regression analysis were used to select genes and to construct a prognosis model. According to the median value of the model's risk score, patients were divided into high and low risk groups, and gene set enrichment analysis (GSEA), immune infiltration and immunotherapy effect, drug resistance and the expression of immune checkpoint related genes and synthetic driver genes of T cell proliferation were analyzed. Finally, the mRNA and protein levels of four genes in PAAD were verified by the clinical proteomic tumor analysis consortium (CPTAC) database and the immunostaining of patients' tissue. 55 differentially expressed oxidative stress genes were identified, and four genes including MET, FYN, CTTN and CDK1 were selected to construct a prognosis model. GESA indicated that immune related pathways, metabolic pathways and DNA repair pathways were significantly enriched in the high risk group as compared to the low risk group. The frequency of genetic mutations was also significantly higher in high risk groups than that in low risk groups. Moreover, the infiltration level of 23 immune cells as well as the expression of immune checkpoint related and synthetic driver genes of T cell proliferation were significantly altered, with the better immunotherapy effect occurring in low risk group. In patient PAAD tissues, the mRNA and protein levels of these four genes were up-regulated. We have successfully constructed a four oxidative stress gene prognostic model that has important predictive value for PAAD patients, and this model might be a promising guidance for prognostic prediction and efficacy monitoring in clinical individualized therapy.

Identification and validation of oxidative stress and immune-related hub genes in Alzheimer’s disease through bioinformatics analysis

Alzheimer’s disease (AD) is the leading cause of dementia in aged population. Oxidative stress and neuroinflammation play important roles in the pathogenesis of AD. Investigation of hub genes for the development of potential therapeutic targets and candidate biomarkers is warranted. The differentially expressed genes (DEGs) in AD were screened in GSE48350 dataset. The differentially expressed oxidative stress genes (DEOSGs) were analyzed by intersection of DEGs and oxidative stress-related genes. The immune-related DEOSGs and hub genes were identified by weighted gene co-expression network analysis (WGCNA) and protein–protein interaction (PPI) analysis, respectively. Enrichment analysis was performed by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes. The diagnostic value of hub genes was assessed by receiver operating characteristic analysis and validated in GSE1297. The mRNA expression of diagnostic genes was determined by qRT-PCR analysis. Finally, we constructed the drug, transcription factors (TFs), and microRNA network of the diagnostic genes. A total of 1160 DEGs (259 up-regulated and 901 down-regulated) were screened in GSE48350. Among them 111 DEOSGs were identified in AD. Thereafter, we identified significant difference of infiltrated immune cells (effector memory CD8 T cell, activated B cell, memory B cell, natural killer cell, CD56 bright natural killer cell, natural killer T cell, plasmacytoid dendritic cell, and neutrophil) between AD and control samples. 27 gene modules were obtained through WGCNA and turquoise module was the most relevant module. We obtained 66 immune-related DEOSGs by intersecting turquoise module with the DEOSGs and identified 15 hub genes through PPI analysis. Among them, 9 hub genes (CCK, CNR1, GAD1, GAP43, NEFL, NPY, PENK, SST, and TAC1) were identified with good diagnostic values and verified in GSE1297. qRT-PCR analysis revealed the downregulation of SST, NPY, GAP43, CCK, and PENK and upregulation of NEFL in AD. Finally, we identified 76 therapeutic agents, 152 miRNAs targets, and 91 TFs regulatory networks. Our study identified 9 key genes associated with oxidative stress and immune reaction in AD pathogenesis. The findings may help to provide promising candidate biomarkers and therapeutic targets for AD.

Alterations in cell viability, reactive oxygen species production, and modulation of gene expression involved in mitogen-activated protein kinase/extracellular regulating kinase signaling pathway by glyphosate and its commercial formulation in hepatocellular carcinoma cells.

Glyphosate (N-phosphonomethyl glycine) is a non-selective, organophosphate herbicide widely used in agriculture and forestry. We investigated the possible toxic effects of the glyphosate active compound and its commercial formulation (Roundup Star®) in the human hepatocellular carcinoma (HepG2) cell line, including their effects on the cytotoxicity, cell proliferation, reactive oxygen species (ROS) levels, and expression of oxidative stress-related genes such as HO-1, Hsp70 Nrf2, L-FABP, and Keap1. MTT and NRU tests indicated that the IC50 values of Roundup Star® were 219 and 140μM, respectively, and because glyphosate failed to induce cell death at the studied concentrations, an IC50 value could not be determined for this cell line. Roundup Star at concentrations of 50 and 100μM significantly increased (39.58% and 52%, respectively) cell proliferation, which 200μM of glyphosate increased by 35.38%. ROS levels increased by 27.97% and 44.77% for 25 and 100μM of Roundup Star and 32.74% and 38.63% for 100 and 200μM of glyphosate exposure. In conclusion, Roundup Star and glyphosate significantly increased expression levels of selected genes related to the mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) signaling pathway. This suggests that ROS production and the MAPK/ERK signaling pathway may be key molecular mechanisms in the toxicity of glyphosate in liver cells.