scholarly journals Alterations of Suckling Piglet Jejunal Microbiota Due to Infection With Porcine Epidemic Diarrhea Virus and Protection Against Infection by Lactobacillus salivarius

2021 ◽  
Vol 8 ◽  
Author(s):  
Wanting Dong ◽  
Ning Ding ◽  
Yu Zhang ◽  
Zhen Tan ◽  
Xiangdong Ding ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Signaling Pathway ◽  
Intestinal Microbiota ◽  
Porcine Epidemic Diarrhea Virus ◽  
Akt Signaling ◽  
Jejunal Mucosa ◽  
Akt Signaling Pathway ◽  
Lactobacillus Salivarius ◽  
Diarrhea Virus ◽  
Porcine Epidemic Diarrhea

The high mortality of neonatal piglets due to porcine epidemic diarrhea virus (PEDV) infection has caused huge economic losses to the pig industry. The intestinal microbiota is an important barrier against invaders entering the gastrointestinal route. In this study, we examined the differences between intestinal microbiota of PEDV-infected and healthy piglets. According to the viral copy numbers, 16 crossbred (Landrace-Yorkshire) piglets were divided into three groups: uninfected, low virus load, and high virus load groups. Next, 16S rRNA sequencing was performed to determine the microbiota composition in jejunal content and jejunal mucosal samples from the three groups. PEDV infection induced an imbalance in the microbiota of both jejunal content and jejunal mucosa. The abundance of phylum Firmicutes was higher in uninfected piglets than in infected piglets, whereas the abundance of Proteobacteria was lower in uninfected piglets. Principal coordinate analysis showed significant separation of jejunal microbiota between different groups. Linear discriminant analysis (LDA) effect size (LEfSe) identified Lactobacillus salivarius as a potential biomarker among three groups at the level of species. Then, in vitro, L. salivarius was able to suppress the infection of PEDV to IPEC-J2 cells and decreased the expression of GRP78 (Glucose-regulating protein 78). In addition, we detected the mRNA expression of genes involved in the FAK/PI3K/Akt signaling pathway. When IPEC-J2 cells were treated with L. salivarius before PEDV infection, the mRNA expression levels of ITGA1, ITGA5, ITGB5, FAK, PIK3R1, PIK3CA and AKT1 were significantly higher than those in the control cells (without treatment) at different times post-infection, indicating that L. salivarius may upregulate the FAK/PI3K/Akt signaling pathway in IPEC-J2 cells to resist PEDV infection. In summary, PEDV infection altered microbial communities in both jejunal content and jejunal mucosa. L. salivarius has a protective effect against PEDV infection in IPEC-J2 cells. This study provides a potentially effective strategy to prevent the occurrence and control the spread of PED in the pig production.

scholarly journals The Antagonistic Effect of Glutamine on Zearalenone-Induced Apoptosis via PI3K/Akt Signaling Pathway in IPEC-J2 Cells

Toxins ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 891
Author(s):  
Tianhu Wang ◽  
Jingjing Wang ◽  
Tong Zhang ◽  
Aixin Gu ◽  
Jianping Li ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Signaling Pathway ◽  
Nuclear Fission ◽  
Akt Signaling ◽  
Antagonistic Effect ◽  
Protective Effects ◽  
Akt Signaling Pathway ◽  
Pi3k Inhibitor Ly294002 ◽  
Apoptotic Genes ◽  
Induced Apoptosis

Zearalenone (ZEN) is a non-steroidal estrogen mycotoxin produced by Fusarium fungi, which inevitably exists in human and animal food or feed. Previous studies indicated that apoptosis seems to be a key determinant of ZEN-induced toxicity. This experiment aimed to investigate the protective effects of Glutamine (Gln) on ZEN-induced cytotoxicity in IPEC-J2 cells. The experimental results showed that Gln was able to alleviate the decline of cell viability and reduce the production of reactive oxygen species and calcium (Ca2+) induced by ZEN. Meanwhile, the mRNA expression of antioxidant enzymes such as glutathione reductase, glutathione peroxidase, and catalase was up-regulated after Gln addition. Subsequently, Gln supplementation resulted in the nuclear fission and Bad-fluorescence distribution of apoptotic cells were weakened, and the mRNA expression and protein expression of pro-apoptotic genes and apoptotic rates were significantly reduced. Moreover, ZEN reduced the phosphorylation Akt, decreased the expression of Bcl-2, and increased the expression of Bax. Gln alleviated the above changes induced by ZEN and the antagonistic effects of Gln were disturbed by PI3K inhibitor (LY294002). To conclude, this study revealed that Gln exhibited significant protective effects on ZEN-induced apoptosis, and this effect may be attributed to the PI3K/Akt signaling pathway.

scholarly journals Acute porcine epidemic diarrhea virus infection reshapes the intestinal microbiota

Virology ◽  
2020 ◽  
Vol 548 ◽  
pp. 200-212
Author(s):  
Shanshan Yang ◽  
Yang Li ◽  
Bin Wang ◽  
Ning Yang ◽  
Xin Huang ◽  
...  
Keyword(s):  
Virus Infection ◽  
Intestinal Microbiota ◽  
Porcine Epidemic Diarrhea Virus ◽  
Diarrhea Virus ◽  
Porcine Epidemic Diarrhea

scholarly journals Porcine Epidemic Diarrhea Virus Induces Vero Cell Apoptosis via the p53-PUMA Signaling Pathway

Viruses ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1218
Author(s):  
Lin Yang ◽  
Chenyu Wang ◽  
Jinqi Shu ◽  
Huapeng Feng ◽  
Yulong He ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Differentially Expressed Genes ◽  
Porcine Epidemic Diarrhea Virus ◽  
Vero Cells ◽  
Pathogenic Mechanism ◽  
Differentially Expressed ◽  
Diarrhea Virus ◽  
Porcine Epidemic Diarrhea ◽  
Induced Apoptosis ◽  
Dependent Pathway

Porcine Epidemic Diarrhea Virus (PEDV) is the causative agent of swine epidemic diarrhea. In order to study the pathogenic mechanism of PEDV, PEDV was inoculated into Vero cells cultured in vitro, and the total RNA of Vero cells was extracted to construct a library for Illumina high-throughput sequencing and screening of differentially expressed genes (p < 0.05). Five differentially expressed genes for qRT-PCR verification analysis were randomly selected, and the verification results were consistent with the transcriptome sequencing results. The Kyoto Encyclopedia of Genes and Genomes (KEGG) signal pathway enrichment analysis was performed on the differentially expressed genes screened above. The results showed that the target gene annotations of differentially expressed genes in the African green monkey genome were mainly enriched in the TNF signaling pathway, the P53 signaling pathway, the Jak-STAT signaling pathway, the MAPK signaling pathway, and immune inflammation. In addition, it has been reported that Puma can promote apoptosis and is a key mediator of P53-dependent and non-dependent apoptosis pathways. However, there is no report that PEDV infection can activate Puma and induce apoptosis in a P53-dependent pathway. It was found by flow cytometry that PEDV infection induced apoptosis, and by Western Blotting detection, PEDV infection significantly increased the expression of p53, BAX, and Puma apoptosis-related proteins. Treatment Vero cells with the p53 inhibitor, PFT-α, could significantly inhibit PEDV-induced apoptosis. Studies have shown that PEDV infection can activate Puma and induce apoptosis in a P53-dependent pathway. These findings provide data support for further elucidating the pathogenic mechanism of PEDV and developing an effective vaccine against PEDV.

scholarly journals TMEM119 facilitates ovarian cancer cell proliferation, invasion, and migration via the PDGFRB/PI3K/AKT signaling pathway

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Tianshui Sun ◽  
Fangfang Bi ◽  
Zhuonan Liu ◽  
Qing Yang
Keyword(s):  
Ovarian Cancer ◽  
Cell Proliferation ◽  
Mrna Expression ◽  
Signaling Pathway ◽  
Akt Signaling ◽  
Biological Behavior ◽  
Akt Signaling Pathway ◽  
Invasion And Migration ◽  
Potential Biomarker ◽  
And Migration

Abstract Background Ovarian cancer (OV) is the deadliest gynecological cancer. Transmembrane protein 119 (TMEM119) has been reported as oncogene in several human cancers. However, the function of TMEM119 in OV is still poorly known. Methods Western blot and qRT-PCR were used to analyze TMEM119 levels. Transwell assays, wound healing assays, CCK-8 assays and EdU cell proliferation assays were designed to explore the function and potential mechanism of TMEM119 in malignant biological behaviors in OV. Results TMEM119 was observed to be overexpressed in OV tissues and associated with poor survival in OV patients. Knockdown and overexpression experiments demonstrated that TMEM119 promoted proliferation, invasion, and migration in OV cells in vitro. TMEM119 mRNA expression was related to the pathways of focal adhesion according to Gene Set Enrichment Analyses and was correlated with the mRNA expression level of platelet-derived growth factor receptor beta (PDGFRB). TMEM119 exerted oncogenic effects partially by regulating the expression of PDGFRB and by activating the PI3K/AKT signaling pathway. Conclusions Collectively, our findings highlight the potential role of TMEM119 in the malignant biological behavior of OV, which may serve as a potential biomarker and a therapeutic candidate for OV.

scholarly journals Differences in the intestinal microbiota between uninfected piglets and piglets infected with porcine epidemic diarrhea virus

PLoS ONE ◽  
2018 ◽  
Vol 13 (2) ◽  
pp. e0192992 ◽  
Author(s):  
Mei-Zhou Huang ◽  
Sheng-Yi Wang ◽  
Hui Wang ◽  
Dong-An Cui ◽  
Ya-Jun Yang ◽  
...  
Keyword(s):  
Intestinal Microbiota ◽  
Porcine Epidemic Diarrhea Virus ◽  
Diarrhea Virus ◽  
Porcine Epidemic Diarrhea

scholarly journals Plastrum Testudinis Extracts Promote BMSC Proliferation and Osteogenic Differentiation by Regulating Let-7f-5p and the TNFR2/PI3K/AKT Signaling Pathway

2018 ◽  
Vol 47 (6) ◽  
pp. 2307-2318 ◽  
Author(s):  
Geng-Yang Shen ◽  
Hui Ren ◽  
Jin-Jing Huang ◽  
Zhi-Da Zhang ◽  
Wen-Hua Zhao ◽  
...  
Keyword(s):  
Protein Expression ◽  
Osteogenic Differentiation ◽  
Mrna Expression ◽  
Signaling Pathway ◽  
Quantitative Polymerase Chain Reaction ◽  
Akt Signaling ◽  
Cell Counting ◽  
Luciferase Reporter ◽  
Akt Signaling Pathway ◽  
Alizarin Red

Background/Aims: Plastrum testudinis extracts (PTE) show osteoprotective effects on glucocorticoid-induced osteoporosis in vivo and in vitro. However, the underlying molecular mechanism of PTE in promoting osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) is unclear. Methods: BMSC proliferation was investigated using the Cell Counting Kit-8 assay. BMSC differentiation and osteogenic mineralization were assayed using alkaline phosphatase and Alizarin red staining, respectively. The mRNA expression levels of Let-7f-5p, Tnfr2, Traf2, Pi3k, Akt, β-catenin, Gsk3β, Runx2, and Ocn were measured using real time quantitative polymerase chain reaction. Protein levels of TNFR2, TRAF2, p-PI3K, p-AKT, p-β-CATENIN, and p-GSK3β were analyzed by western blotting. The functional relationship of Let-7f-5p and Tnfr2 was determined by luciferase reporter assays. Results: The optimum concentration for PTE was 30 μg/ml. PTE significantly promoted BMSC osteogenic differentiation and mineralization after 7 and 14 days in culture, respectively. The combination of PTE and osteogenic induction exhibited significant synergy. PTE upregulated Let-7f-5p, β-catenin, Runx2, and Ocn mRNA expression, and downregulated Tnfr2, Traf2, Pi3k, Akt, and Gsk3β mRNA expression. PTE inhibited TNFR2, TRAF2, and p-β-CATENIN protein expression, and promoted p-PI3K, p-AKT, and p-GSK3β protein expression. In addition, Tnfr2 was a functional target of Let-7f-5p in 293T cells. Conclusions: Our results suggested that PTE may promote BMSC proliferation and osteogenic differentiation via a mechanism associated with the regulation of Let-7f-5p and the TNFR2/PI3K/AKT signaling pathway.

scholarly journals N-CoR modulates osteogenic differentiation of rat mesenchymal stem cells through the PI3K/Akt-cell signaling pathway

2016 ◽  
Vol 68 (3) ◽  
pp. 551-559
Author(s):  
Yi Qin ◽  
Guoping Cao ◽  
Jichao Ye ◽  
Peng Wang ◽  
Liangbin Gao ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Protein Expression ◽  
Osteogenic Differentiation ◽  
Mrna Expression ◽  
Signaling Pathway ◽  
Akt Signaling ◽  
Akt Pathway ◽  
Akt Signaling Pathway ◽  
Alp Activity

The nuclear receptor corepressor (N-CoR) is involved in the regulation of diverse transcription factors. We previously found that N-CoR could regulate adipogenic differentiation of rat mesenchymal stem cells (MSCs),but whether it modulated osteogenic differentiation of this type of cells was uncertain. Therefore, in the present study, we investigated the effect and mechanism of N-CoRon osteogenic differentiation of rat MSCs. The results showed that MSCs cultured in osteogenic medium successfully differentiated into osteogenic cells. Overexpression of N-CoR decreased cell proliferation, alkaline phosphatase (ALP)activity, calcium accumulation, mRNA expression of genes including bone sialoprotein (BSP), osteocalcin (OCN), osteopontin (OPN), Osterix and Runx2, and protein expression of phosphor-Akt(pAkt). Conversely, knocking down cellular N-CoR by small interfering RNA (siRNA) promoted pAkt activity and cell differentiation. Overexpression or knockdown of N-CoRhad no significant influences on the protein expression of pyruvate dehydrogenase kinase isozyme 1 (PDK1), phosphatidylinositol 3-kinase (PI3K) and total Akt, indicating that N-CoR regulated the changes in the PI3K/Akt signaling pathway by targeting pAkt. To further prove the function of the PI3K/Akt signaling in N-CoR-regulated osteogenic differentiation, we used the PI3K inhibitor (LY294002) to block the activation of the PI3K/Akt pathway and found that overexpression of N-CoR showed no effects on ALP activity, calcium level and mRNA expression of BSP, osteocalcin OCN, OPN, Osterix and Runx2 in rat MSCs following the inhibition of the PI3K/Akt pathway. These findings suggest that N-CoR regulates osteogenic differentiation of rat MSCs through suppression of the PI3K/Akt signaling pathway.

scholarly journals Identification of the Suitable Insertion Site for Expression of Spike Gene of A Porcine Epidemic Diarrhea Virus Variant in A Pseudorabies Virus Vector

2020 ◽  
Author(s):  
Chuanjian Zhang ◽  
Shiqi Guo ◽  
Rongli Guo ◽  
Saisai Chen ◽  
Yating Zheng ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Post Infection ◽  
Porcine Epidemic Diarrhea Virus ◽  
Pseudorabies Virus ◽  
Insertion Site ◽  
Diarrhea Virus ◽  
Artificial Chromosomes ◽  
S Gene ◽  
Porcine Epidemic Diarrhea ◽  
Gene Mrna

Abstract Background: The emergence of variant porcine epidemic diarrhea virus (PEDV) strain and pseudorabies virus (PRV) in China in recent years has decreased the effectiveness of CV777 and Bartha K61 vaccines, causing significant loss to the swine industry. Previously, we generated a TK&gE-deleted PRV (PRVTK&gE-AH02) based on a virulent PRV AH02LA strain. It was shown to be safe for 1-day-old piglets with maternal PRV antibodies and 4～5 week-old PRV antibody negative piglets and provide rapid and complete protection in weaned pigs against lethal challenge with the PRV variant strain. PRVTK&gE-AH02 may be a promising live vaccine vector for construction of recombinant vaccine in pigs. PEDV spike (S) protein is mainly used in the development of PEDV vaccines. Therefore, the gene-deleted PRV (from PRV variants) vectored vaccine expressing variant PEDV S gene may be viable PEDV and PRV vaccine candidates. However, insertion site is an important factor affecting foreign gene expression and vaccine efficacy. Results: In this study, we constructed four recombinant PRV-S bacterial artificial chromosomes (BACs) carrying the same S expression cassette in different noncoding regions (UL11-10, UL35-36, UL46-27 or US2-1) from AH02LA BAC with TK, gE and gI deletion. The successful expression of S gene (UL11-10, UL35-36 and UL46-27) in recombinant viruses was confirmed by virus rescue, PCR, RT-PCR and indirect immunofluorescence. We observed higher S gene mRNA expression level in Swine testicular cells infected with PRV-S(UL11-10)ΔTK/gE and PRV-S (UL35-36)ΔTK/gE compared to that of PRV-S(UL46-27)ΔTK/gE at 6 hour post infection (P < 0.05). Moreover, at 12 hour post infection, cells infected with PRV-S (UL11-10)ΔTK/gE exhibited higher S gene mRNA expression than those infected with PRV-S (UL35-36)ΔTK/gE (P = 0.097) and PRV-S (UL46-27)ΔTK/gE (P < 0.05). Recovered vectored mutant PRV-S (UL11-10, UL35-36 and UL46-27) exhibited similar growth kinetics to the parental virus (PRVTK&gE-AH02).Conclusions: The identification and comparison of the insertion sites in PRV genome laids a foundation for future development of recombinant PRV vaccines.

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