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1.
Genes (Basel) ; 11(4)2020 Mar 26.
Artigo em Inglês | MEDLINE | ID: mdl-32224871

RESUMO

Heme oxygenase 1 (HMOX1) is a stress-inducing enzyme with multiple cardiovascular protective functions, especially in hypoxia stress. However, transcriptional regulation of swine HMOX1 gene remains unclear. In the present study, we first detected tissue expression profiles of HMOX1 gene in adult Hezuo Tibetan pig and analyzed the gene structure. We found that the expression level of HMOX1 gene was highest in the spleen of the Hezuo Tibetan pig, followed by liver, lung, and kidney. A series of 5' deletion promoter plasmids in pGL3-basic vector were used to identify the core promoter region and confirmed that the minimum core promoter region of swine HMOX1 gene was located at -387 bp to -158 bp region. Then we used bioinformatics analysis to predict transcription factors in this region. Combined with site-directed mutagenesis and RNA interference assays, it was demonstrated that the three transcription factors WT1, Sp1 and C/EBPα were important transcription regulators of HMOX1 gene. In summary, our study may lay the groundwork for further functional study of HMOX1 gene.

2.
Microb Pathog ; 136: 103699, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31472261

RESUMO

Clostridium perfringens (C. perfringens) type C is one of major pathogenic causing diarrhea and other intestinal inflammatory diseases in piglets, which seriously affects the healthy development of the swine industries. Studies have found that miRNAs play important roles in regulating piglet diarrhea challenged by pathogenic E. coli and Salmonella. However, little is known miRNAs in the ileum of diarrheic piglets caused by C. perfringens type C. Therefore, we studied the expression profiles of the ileum miRNAs of 7-day-old piglets infected with C. perfringens type C using small RNA-Seq, including control (IC), susceptible (IS) and resistant (IR) groups. As a result, 53 differentially expressed miRNAs were found. KEGG pathway analysis for target genes revealed that these miRNAs were involved in ErbB signaling pathway, MAPK signaling pathway, Jak-STAT signaling pathway and Wnt signaling pathway. The expression correlation analysis between miRNAs and target genes revealed that the expression of miR-7134-5p had negative correlation with target NFATC4, miR-500 had negative correlation with target ELK1, HSPA2 and IL7R, and miR-92b-3p had negative correlation with target CLCF1 in ileum of IR vs IS group, suggesting that miR-7134-5p targeting to NFATC4, miR-500 targeting to ELK1, HSPA2 and IL7R, and miR-92b-3p targeting to CLCF1 were probably involved in piglet resisting C. perfringens type C. The results will provide value resources for better understanding of the genetic basis of C. perfringens type C resistance in piglet and lays a new foundation for identifying novel markers of C. perfringens type C resistance.

3.
Microbiologyopen ; 8(12): e923, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31496126

RESUMO

Diarrhea is one of the most common enteric diseases in young piglets. Diverse factors such as an unstable gut microenvironment, immature intestinal immune system, early supplementary feeding, and weaning often induce dysfunction of gut microbiota, thus leading to a continuing high incidence of diarrhea in piglets. However, few studies have characterized the gut microbiota of diarrheic piglets following changes in diet and during the development of intestinal physiology. In this study, we used 16S rRNA gene sequencing to analyze the dynamic establishment of fecal microbiota in six healthy piglets in response to age-related changes in the diet: sow-reared, early supplementary creep-feeding (sow-reared + starter diet), and weaning (solid nursery diet). We compared the gut microbiota of these six healthy piglets with those of diarrheic piglets during each of the three dietary stages (n = 10 sow-reared, n = 10 early supplementary creep-feeding, and n = 5 weaning). We found that weaning (solid nursery feeding) was the primary factor leading to dynamic colonization by microbiota in healthy piglets, and diarrhea primarily affected the microbial communities of piglets before weaning. Healthy piglets showed a continuous decrease in Lactobacillus and Escherichia, as well as a gradual increase in Prevotella with the transition to solid food. An altered relationship between Prevotella and Escherichia may be the main cause of diarrhea in preweaned piglets, whereas reduced numbers of Bacteroides, Ruminococcus, Bulleidia, and Treponema that are responsible for the digestion and utilization of solid feeds may be related to the onset of postweaning piglet diarrhea. The Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) functional analysis indicated that a reduction in genes involved in carbohydrate metabolism induced by intestinal dysbacteriosis in diarrheic piglets was one of the major causes of diarrhea at the three dietary stages. These findings provide insights into developing an intervention strategy for better management of diarrhea in piglets.

4.
Exp Ther Med ; 18(3): 2160-2166, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31452707

RESUMO

Lipodystrophic patients have an adipose tissue triglyceride storage defect that causes ectopic lipid accumulation, leading to severe insulin resistance. The present study investigated the potential role of isoimperatorin on 3T3-L1 adipocyte differentiation. mRNA and protein levels of differentiation- and lipid accumulation-associated genes, as well as the adipogenesis-related signaling pathway were analyzed in control and isoimperatorin-treated differentiated 3T3-L1 adipocytes using reverse transcription-quantitative PCR and western blot analysis. Results determined that isoimperatorin promoted 3T3-L1 fibroblast adipogenesis in a dose-dependent manner compared with standard differentiation inducers. Isoimperatorin significantly increased mRNA and protein expression of the crucial adipogenic transcription factors peroxisome proliferator activated receptor-γ (PPARγ) and CCAAT enhancer binding protein-α (C/EBPα). mRNA expression of the downstream adipogenesis-related genes sterol regulatory element-binding transcription factor 1c, adipocyte protein 2, fatty acid synthase, adiponectin and diacylglycerol O-acyltransferase 2 were also significantly increased following isoimperatorin treatment. The underlying mechanism likely involved activation of the Akt signaling pathway. Taken together, the present findings indicated that isoimperatorin may alter PPARγ and C/EBPα expression via the Akt signaling pathway, resulting in promotion of adipogenesis. The results highlighted the potential use of isoimperatorin as a therapeutic agent for preventing diabetes.

5.
Microb Pathog ; 135: 103567, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31163250

RESUMO

Clostridium perfringens (C. perfringens), a Gram-positive bacterium, is one of the main causing piglet diarrhea, which leads serious economic loss in the world swine industries. Generally, the innate immune response plays a critical role in host defense against pathogen invasion. TLR4, a member of the TLR (Toll-like receptor) family, has been considered to implicate in the host immune responses and induce secretion of inflammatory cytokines during bacterial infection. However, little is clear about the effects of TLR4 and key signaling genes in the process of piglet inflammatory and immune responses after C. perfringens infection. This study aims to explore the effect of C. perfringens type C infection on the key mRNAs of TLR4/MyD88/NF-κB signaling pathways during the process of piglet diarrhea. In this study, the expressions of TLR4 and other key mRNAs in the TLR4/MyD88/NF-κB signaling pathways were quantified in piglet ileum and jejunum tissues among IR (intestinal resistance), IS (intestinal susceptibility) and IC (intestinal control) groups by qPCR and Western blot methods, the concentrations of pro-inflammatory cytokines in intestinal tissues and serum immunoglobulins were also tested by ELISA kits. Results showed that compared to IC group, expressions of ileum TLR4 and TNF-α was significantly increased in the IS and IR groups, specially TBK1 gene; the expressions of ileum TLR2, TRAF6, MyD88 and IL-8 mRNAs was significantly up-regulated in the IS group, the expressions of TLR9, NF-κB, IL-6, IFN-γ and MAPK1 genes were not significant differences among the IR, IS and IC groups. Meanwhile, the protein levels of TLR4, HMGB1 and NF-κB were higher in the IS and IR groups. The levels of jejunum IFN-γ and IL-6, ileum IL-6 and IL-12 were risen in the IR group. Serum immunoglobulin IgA and IgG in the IR and IS groups reached a peak on the 72 h and 48 h post infection, respectively. These findings suggest that C. perfringens type C infection induces host immune responses involving in the TLR4/MyD88/NF-κB signaling pathways in ileum than in jejunum, which may provide valuable information for innate immune mechanisms involved in regulation of piglet diarrhea caused by C. perfringens type C infection.


Assuntos
Infecções por Clostridium/imunologia , Clostridium perfringens/patogenicidade , Intestino Delgado/imunologia , Fator 88 de Diferenciação Mieloide/metabolismo , NF-kappa B/metabolismo , Transdução de Sinais , Receptor 4 Toll-Like/metabolismo , Animais , Infecções por Clostridium/microbiologia , Citocinas/genética , Citocinas/metabolismo , Diarreia/imunologia , Diarreia/microbiologia , Modelos Animais de Doenças , Regulação da Expressão Gênica , Imunidade Inata , Imunoglobulinas/sangue , Intestino Delgado/microbiologia , Fator 88 de Diferenciação Mieloide/genética , NF-kappa B/genética , Suínos , Receptor 4 Toll-Like/genética , Receptores Toll-Like/genética , Receptores Toll-Like/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
6.
Animals (Basel) ; 9(5)2019 May 23.
Artigo em Inglês | MEDLINE | ID: mdl-31126046

RESUMO

C. perfringens type C can induce enteritis accompanied by diarrhea and annually causes significant economic losses to the global pig industry. The pathogenic mechanisms of C. perfringens type C in pigs are still largely unknown. To investigate this, we challenged seven-day-old piglets with C. perfringens type C to cause diarrhea. We performed hematoxylin & eosin (H&E) staining of the small intestine (including duodenum, jejunum, and ileum) and assessed gene expression in the ileal tissue. H&E staining of the duodenum, jejunum, and ileum demonstrated inflammation and edema of the lamina propria and submucosa. A total of 2181 differentially expressed genes (DEGs) were obtained in ileal tissues. Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis of DEGs indicated that the main pathways were enriched in the T cell receptor signaling pathway, NF-kappa B signaling pathway, and (tumor necrosis factor) TNF signaling pathway. These results provide insights into the pathogenicity of C. perfringens type C and improve our understanding of host-bacteria interactions.

7.
Artigo em Inglês | MEDLINE | ID: mdl-31114763

RESUMO

Long non-coding RNAs (lncRNAs) have been shown to play important roles in regulating host immune and inflammatory responses to bacterial infection. Infection with Clostridium perfringens (C. perfringens), a food-borne zoonotic pathogen, can lead to a series of inflammatory diseases in human and piglet, greatly challenging the healthy development of global pig industry. However, the roles of lncRNAs involved in piglet immune response against C. perfringens type C infection remain unknown. In this study, the regulatory functions of ileum lncRNAs and mRNAs were investigated in piglet immune response to C. perfringens type C infection among resistance (IR), susceptibility (IS) and sham-inoculation (control, IC) groups. A total of 480 lncRNAs and 3,669 mRNAs were significantly differentially expressed, the differentially expressed lncRNAs and mRNAs in the IR and IS groups were enriched in various pathways of ABC transporters, olfactory transduction, PPAR signaling pathway, chemokine signaling pathway and Toll-like receptor signaling pathway, involving in regulating piglet immune responses and resistance during infection. There were 212 lncRNAs and 505 target mRNAs found to have important association with C. perfringens infectious diseases, furthermore, 25 dysregulated lncRNAs corresponding to 13 immune-related target mRNAs were identified to play potential roles in defense against bacterial infection. In conclusion, the results improve our understanding on the characteristics of lncRNAs and mRNAs on regulating host immune response against C. perfringens type C infection, which will provide a reference for future research into exploring C. perfringens-related diseases in human.


Assuntos
Infecções por Clostridium/veterinária , Clostridium perfringens/imunologia , Regulação da Expressão Gênica , Íleo/imunologia , RNA Longo não Codificante/metabolismo , RNA Mensageiro/metabolismo , Doenças dos Suínos/imunologia , Animais , Animais Recém-Nascidos , Infecções por Clostridium/imunologia , Infecções por Clostridium/patologia , Íleo/patologia , Suínos , Doenças dos Suínos/patologia
8.
Fish Shellfish Immunol ; 89: 516-524, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30986537

RESUMO

Infectious hematopoietic necrosis virus (IHNV) leads to serious disease and economic losses in the salmonid aquaculture industry. The present study aimed to develop an effective and efficient vaccine to protect rainbow trout (Oncorhynchus mykiss) against IHNV infection. Administered via the immersion route, a live vector vaccine containing the regions of the IHNV glycoprotein (G) induced immune responses in rainbow trout. Use of the immersion route induced more-efficient mucosal immunity than intramuscular injection vaccination. IHNV G gene expression was detected in the spleens of rainbow trout at 3, 7 and 15 days post-vaccination (dpv). The G gene expression continuously decreased between 3 and 15 dpv. In addition, the expression of TLR-3, TLR-7 and TLR-8 was upregulated after vaccination, and the highest expression levels of IFN-1, Mx-1, Mx-3, Vig-1 and Vig-2 were observed at 3 dpv. Four markers of the adaptive immune response (CD4, CD8, IgM and IgT) gradually increased. When experimental fish were challenged with IHNV by immersion, significant differences in cumulative percentage mortality were observed in the vaccinated fish and the unvaccinated (empty-plasmid-vaccinated) fish. The relative survival rate was 92% and 6% in the vaccinated group and empty-plasmid group, respectively. Serum antibody levels gradually increased in the vaccinated fish, unlike in the unvaccinated fish, after 7 dpv. Our results suggest there was a significant increase in fish immune responses and resistance to infection with IHNV following administration of the live vector vaccine. Therefore, this live vector vaccine is a promising vaccine that may be utilized to protect rainbow trout against IHNV.


Assuntos
Imunidade Adaptativa , Doenças dos Peixes/prevenção & controle , Vírus da Necrose Hematopoética Infecciosa/fisiologia , Oncorhynchus mykiss , Infecções por Rhabdoviridae/veterinária , Vacinas Virais/imunologia , Animais , Doenças dos Peixes/imunologia , Glicoproteínas/genética , Glicoproteínas/imunologia , Infecções por Rhabdoviridae/imunologia , Infecções por Rhabdoviridae/prevenção & controle , Baço/imunologia , Vacinas Atenuadas/imunologia , Proteínas Virais/genética , Proteínas Virais/imunologia
9.
PeerJ ; 6: e5997, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30533301

RESUMO

Background: Clostridium perfringens (C. perfringens) type C is the most common bacteria causing piglet diarrheal disease and it greatly affects the economy of the global pig industry. The spleen is an important immune organ in mammals; it plays an irreplaceable role in resisting and eradicating pathogenic microorganisms. Based on different immune capacity in piglets, individuals display the resistance and susceptibility to diarrhea caused by C. perfringens type C. Recently, long non-coding RNA (lncRNA) and mRNA have been found to be involved in host immune and inflammatory responses to pathogenic infections. However, little is known about spleen transcriptome information in piglet diarrhea caused by C. perfringens type C. Methods: Hence, we infected 7-day-old piglets with C. perfringens type C to lead to diarrhea. Then, we investigated lncRNA and mRNA expression profiles in spleens of piglets, including control (SC), susceptible (SS), and resistant (SR) groups. Results: As a result, 2,056 novel lncRNAs and 2,417 differentially expressed genes were found. These lncRNAs shared the same characteristics of fewer exons and shorter length. Bioinformatics analysis identified that two lncRNAs (ALDBSSCT0000006918 and ALDBSSCT0000007366) may be involved in five immune/inflammation-related pathways (such as Toll-like receptor signaling pathway, MAPK signaling pathway, and Jak-STAT signaling pathway), which were associated with resistance and susceptibility to C. perfringens type C infection. This study contributes to the understanding of potential mechanisms involved in the immune response of piglets infected with C. perfringens type C.

10.
FEBS Open Bio ; 8(10): 1722-1732, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30338222

RESUMO

Clostridium perfringens type C is a pathogen that causes necrotizing enteritis (NE), which is an intestinal tract disease in piglets. The pathogenesis of C. perfringens type C-induced NE is still unclear, leading to a lack of effective therapies. Earlier studies have reported that circular RNAs (circRNAs) are involved in the pathogenic processes of various diseases. However, it is not known if circRNAs in spleen play a role in C. perfringens type C infection in NE. To address this question, we infected 7-day-old piglets with C. perfringens type C to induce NE. Hematoxylin and eosin staining of small intestine revealed inflammation, atrophy and shedding of intestinal villi, and intestinal mucosal necrosis. We observed increased expression of cytokine genes (such as IL-1ß and IL-6) and inflammation in the spleen. In addition, we used RNA-seq and bioinformatics analysis to examine changes in circRNA expression. A total of 103 circRNAs were found to be differentially expressed in NE, and Gene Ontology analysis revealed that the genes producing differentially expressed circRNAs were enriched in regulation of the cellular metabolic process protein binding. Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that the genes producing differentially expressed circRNAs were involved in the tumor necrosis factor signaling pathway, T cell receptor signaling pathway and nuclear factor-κB signaling pathway. Finally, we found eight circRNAs (including circ_0002220 and circ_0000821) that are related to NE. Therefore, our study provides new insights into the mechanisms underlying C. perfringens type C infection in piglets.

11.
PLoS One ; 12(5): e0178045, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28542522

RESUMO

MicroRNAs (miRNAs) are small non-coding RNAs that are involved in translational regulation of the messenger RNA molecules. Sequence variations in the genes encoding miRNAs could influence their biogenesis and function. MiR-15b plays an important role in cellular proliferation, apoptosis and the cell cycle. Here, we report the identification of a C58T mutation in porcine pre-miR-15b. Through in vitro and in vivo experiments, we determined that this mutation blocks the transition from pri-miRNA to pre-miRNA, alters the strand selection between miR-15b-5p and miR-15b-3p, and obstructs biogenesis of the downstream miR-16-1. These results serve to highlight the importance of miRNA mutations and their impacts on miRNA biogenesis.


Assuntos
MicroRNAs/biossíntese , MicroRNAs/genética , Precursores de RNA/genética , Animais , Linhagem Celular , Regulação da Expressão Gênica , Células HEK293 , Humanos , MicroRNAs/ultraestrutura , Mutação/genética , Precursores de RNA/metabolismo , Estabilidade de RNA/genética , Sus scrofa
12.
Front Microbiol ; 8: 502, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28392784

RESUMO

Diarrhea is a leading cause of increased mortality in neonatal and young piglets. Aberration of the gut microbiota is one important factor in the etiology of piglet diarrhea. However, information regarding the structure and function of the gut microbiome in diarrheic neonatal piglets is limited. To investigate the composition and functional potential of the fecal microbiota in neonatal piglets, we performed 16S rRNA gene sequencing on 20 fecal samples from diarrheic piglets and healthy controls, and metagenomics sequencing on a subset of six samples. We found striking compositional and functional differences in fecal microbiota between diarrheic and healthy piglets. Neonatal piglet diarrhea was associated with increases in the relative abundance of Prevotella, Sutterella, and Campylobacter, as well as Fusobacteriaceae. The increased relative abundance of Prevotella was correlated with the reduction in Escherichia coli and the majority of beneficial bacteria that belonging to the Firmicutes phylum (e.g., Enterococcus, Streptococcus, Lactobacillus, Clostridium, and Blautia) in diarrheic piglets. The differentially functional gene abundances in diarrheic piglets were an increase in bacterial ribosome, and contributed primarily by the genera Prevotella, this indicates a growth advantage of the Prevotella in diarrheic conditions. Additional functional gene sets were associated with the reduction of polyamine transport, monosaccharide and sugar-specific PTS transport, amino acid transport, and two-component regulatory system. These profiles likely impact the ability to transport and uptake nutrients, as well as the ability to fight microbial infections in the piglet gut ecosystem. This work identifies a potential role for Prevotella in the community-wide microbial aberration and dysfunction that underpins the pathogenesis of piglet diarrhea. Identification of these microbial and functional signatures may provide biomarkers of neonatal piglet diarrhea.

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