RESUMO
Porcine reproductive and respiratory syndrome (PRRS) is one of the most economically devastating diseases affecting the swine industry worldwide. To investigate the role of miRNAs in the infection and susceptibility of PRRS virus (PRRSV), twenty-four miRNA libraries were constructed and sequenced from PRRSV-infected and mock-infected Porcine alveolar macrophages (PAMs) of Meishan, Landrace, Pietrain and Qingping pigs at 9 hours post infection (hpi), 36 hpi, and 60 hpi. The let-7 family miRNAs were significantly differentially expressed between PRRSV-infected and mock-infected PAMs from 4 pig breeds. The let-7 family miRNAs could significantly inhibit PRRSV-2 replication by directly targeting the 3'UTR of the PRRSV-2 genome and porcine IL6, which plays an important role in PRRSV replication and lung injury. NEAT1 acts as a competing endogenous lncRNA (ceRNA) to upregulate IL6 by attaching let-7 in PAMs. EMSA and ChIP results confirmed that ARID3A could bind to the promoter region of pri-let-7a/let-7f/let-7d gene cluster and inhibit the expression of the let-7 family. Moreover, the NF-κB signaling pathway inhibits the expression of the let-7 family by affecting the nuclear import of ARID3A. The pEGFP-N1-let-7 significantly reduced viral infections and pathological changes in PRRSV-infected piglets. Taken together, NEAT1/ARID3A/let-7/IL6 play significant roles in PRRSV-2 infection and may be promising therapeutic targets for PRRS.
Assuntos
MicroRNAs , Síndrome Respiratória e Reprodutiva Suína , Vírus da Síndrome Respiratória e Reprodutiva Suína , RNA Longo não Codificante , Regiões 3' não Traduzidas , Animais , Proteínas de Ligação a DNA/genética , Interleucina-6/metabolismo , Macrófagos Alveolares/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , NF-kappa B/genética , NF-kappa B/metabolismo , Síndrome Respiratória e Reprodutiva Suína/genética , Síndrome Respiratória e Reprodutiva Suína/metabolismo , Vírus da Síndrome Respiratória e Reprodutiva Suína/metabolismo , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Suínos , Fatores de Transcrição/genética , Replicação ViralRESUMO
Porcine reproductive and respiratory syndrome (PRRS), a highly contagious disease caused by Porcine reproductive and respiratory syndrome virus (PRRSV), results in huge economic losses to the world pig industry. MiRNAs have been reported to be involved in regulation of viral infection. In our study, miR-320 was one of 21 common differentially expressed miRNAs of Meishan, Pietrain, and Landrace pig breeds at 9-h post-infection (hpi). Bioinformatics and experiments found that PRRSV replication was inhibited by miR-320 through directly targeting PRRSV ORF6. In addition, the expression of CCAAT enhancer binding protein beta (CEBPB) was also inhibited by miR-320 by targeting the 3' UTR of CEBPB, which significantly promotes PRRSV replication. Intramuscular injection of pEGFP-N1-miR-320 verified that miR-320 significantly inhibited the replication of PRRSV and alleviated the symptoms caused by PRRSV in piglets. Taken together, miR-320 have significant roles in the infection and may be promising therapeutic target for PRRS.
Assuntos
MicroRNAs , Síndrome Respiratória e Reprodutiva Suína , Vírus da Síndrome Respiratória e Reprodutiva Suína , Proteínas Virais , Replicação Viral , Animais , Proteína beta Intensificadora de Ligação a CCAAT/metabolismo , Proteína beta Intensificadora de Ligação a CCAAT/genética , MicroRNAs/genética , MicroRNAs/metabolismo , Síndrome Respiratória e Reprodutiva Suína/genética , Síndrome Respiratória e Reprodutiva Suína/virologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Vírus da Síndrome Respiratória e Reprodutiva Suína/fisiologia , Suínos , Proteínas Virais/genética , Proteínas Virais/metabolismoRESUMO
Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) is one of the major diseases restricting the development of large-scale pig breeding industry, which has brought huge economic losses to pig industry. Although a lot of work has been done in vaccine development, biosafety and pig health, PRRSV is characterized by easy mutation and recombination of genome, immunosuppression, enhanced antibody dependence, persistent infection, etc., making the prevention, control and elimination of PRRSV very difficult. With the deepening of PRRSV research, it is gradually realized that screening and identifying PRRSV susceptibility/resistance genes and implementing PRRSV disease resistance breeding are long-term and effective strategies for fundamental prevention and control, which has important practical significance for the prevention and control of pig herds.
RESUMO
Glycyrrhiza polysaccharide (GCP) is a natural plant active polysaccharide extracted from traditional Chinese medicine licorice. In this research, we studied the antiviral activity of glycyrrhiza polysaccharide against porcine reproductive and respiratory syndrome virus (PRRSV), a virus of the Arteriviridae family, with a high rate of variation and has caused huge economic losses to the pig industry in various countries since its discovery. Our results show that GCP can inhibit PRRSV replication in a dose-dependent manner. Furthermore, GCP could inhibit the mRNA expression of receptor genes CD163 and NF-κB p65 and promote the mRNA expression of the SLA-7 gene. Because of these results, GCP can be used as a candidate drug to prevent and treat PRRS.
Assuntos
Glycyrrhiza , Síndrome Respiratória e Reprodutiva Suína , Vírus da Síndrome Respiratória e Reprodutiva Suína , Suínos , Animais , Linhagem Celular , RNA Mensageiro , Replicação ViralRESUMO
BACKGROUND: MicroRNAs (miRNAs) are small non-coding RNAs playing vital roles in regulating posttranscriptional gene expression. Elucidating the expression regulation of miRNAs underlying pig testis development will contribute to a better understanding of boar fertility and spermatogenesis. RESULTS: In this study, miRNA expression profile was investigated in testes of Duroc and Meishan boars at 20, 75, and 270 days of age by high-throughput sequencing. Forty-five differentially expressed miRNAs were identified from testes of Duroc and Meishan boars before and after puberty. Integrated analysis of miRNA and mRNA profiles predicted many miRNA-mRNA pairs. Gene ontology and biological pathway analyses revealed that predicted target genes of ssc-mir-423-5p, ssc-mir-34c, ssc-mir-107, ssc-mir-196b-5p, ssc-mir-92a, ssc-mir-320, ssc-mir-10a-5p, and ssc-mir-181b were involved in sexual reproduction, male gamete generation, and spermatogenesis, and GnRH, Wnt, and MAPK signaling pathway. Four significantly differentially expressed miRNAs and their predicted target genes were validated by quantitative real-time polymerase chain reaction, and phospholipase C beta 1 (PLCß1) gene was verified to be a target of ssc-mir-423-5p. CONCLUSIONS: This study provides an insight into the functional roles of miRNAs in testis development and spermatogenesis and offers useful resources for understanding differences in sexual function development caused by the change in miRNAs expression between Duroc and Meishan boars.
Assuntos
Redes Reguladoras de Genes , MicroRNAs/genética , RNA Mensageiro/genética , Suínos/genética , Testículo/metabolismo , Transcriptoma , Animais , Linhagem Celular , Masculino , MicroRNAs/metabolismo , Fosfolipase C beta/genética , Fosfolipase C beta/metabolismo , RNA Mensageiro/metabolismo , Transdução de Sinais , Espermatogênese , Suínos/metabolismo , Testículo/citologiaRESUMO
Porcine reproductive and respiratory syndrome (PRRS), caused by the PRRS virus (PRRSV), is an economically significant contagious disease. Traditional approaches based on vaccines or medicines were challenging to control PRRSV due to the diversity of viruses. Different breeds of pigs infected with PRRSV have been reported to have different immune responses. However, due to the complexity of interaction mechanism between host and PRRSV, the genetic mechanism leading to PRRSV susceptibility/resistance in various pig breeds is still unclear. Herein, the role of host genetic components in PRRSV susceptibility is systematically described, and the molecular mechanisms by which host genetic factors such as SNPs, cytokines, receptor molecules, intestinal flora, and non-coding RNAs regulate PRRSV susceptibility/resistance. Therefore, improving the resistance to disease of individual animals through disease-resistance breeding technology is of profound significance for uplifting the sustainable and healthy development of the pig industry.
Assuntos
Síndrome Respiratória e Reprodutiva Suína , Vírus da Síndrome Respiratória e Reprodutiva Suína , Suínos , Animais , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Síndrome Respiratória e Reprodutiva Suína/genética , Citocinas/genéticaRESUMO
Porcine reproductive and respiratory syndrome (PRRS) is an infectious disease caused by the PRRS virus that leads to reproductive disorders and severe dyspnoea in pigs, which has serious economic impacts. One of the reasons PRRSV cannot be effectively controlled is that it has developed countermeasures against the host immune response, allowing it to survive and replicate for long periods. Transcription Factors acts as a bridge in the interactions between the host and PRRSV. PRRSV can create an environment conducive to PRRSV replication through transcription factors acting on miRNAs, inflammatory factors, and immune cells. Conversely, some transcription factors also inhibit PRRSV proliferation in the host. In this review, we systematically described how PRRSV uses host transcription factors such as SP1, CEBPB, STATs, and AP-1 to escape the host immune system. Determining the role of transcription factors in immune evasion and understanding the pathogenesis of PRRSV will help to develop new treatments for PRRSV.
RESUMO
Porcine reproductive and respiratory syndrome (PRRS) caused by a single-stranded RNA virus (PRRSV) is a highly infectious respiratory disease and leads to huge economic losses to the swine industry worldwide. To investigate the role of miRNAs in the infection and lung injury induced by PRRSV, the differentially expressed miRNAs (DE-miRs) were isolated from PRRSV-2 infected/mock-infected PAMs of Meishan, Landrace, Pietrain, and Qingping pigs at 9, 36, and 60 hpi. Mir-331-3p was the only common DE-miR in each set of miRNA expression profile at 36 hpi. Mir-210 was one of 7 common DE-miRs between PRRSV infected and mock-infected PAMs of Meishan, Pietrain, and Qingping pigs at 60 hpi. Mir-331-3p/mir-210 could target PRRSV-2 ORF1b, bind and downregulate porcine TNF-α/STAT1 expression, and inhibit PRRSV-2 replication, respectively. Furthermore, STAT1 and TNF-α could mediate the transcriptional activation of MCP-1, VCAM-1, and ICAM-1. STAT1 could also upregulate the expression of TNF-α by binding to its promoter region. In vivo, pEGFP-N1-mir-331-3p could significantly reduce viral replication and pathological changes in PRRSV-2 infected piglets. Taken together, Mir-331-3p/mir-210 have significant roles in the infection and lung injury caused by PRRSV-2, and they may be promising therapeutic targets for PRRS and lung injury/inflammation.
Assuntos
Pulmão/metabolismo , Pulmão/virologia , MicroRNAs/genética , Síndrome Respiratória e Reprodutiva Suína/genética , Síndrome Respiratória e Reprodutiva Suína/virologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/fisiologia , Replicação Viral , Animais , Biomarcadores , Biópsia , Linhagem Celular , Regulação da Expressão Gênica , Interações Hospedeiro-Patógeno , Pulmão/patologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/classificação , Interferência de RNA , Fator de Transcrição STAT1/genética , Suínos , Fator de Necrose Tumoral alfa/genética , Replicação Viral/genéticaRESUMO
Gonadotropin-releasing hormone 2 receptor (GnRHR2) together with its cognate ligand involves in regulating reproductive behavior. However, little is known concerning the effect of transcription factor steroidogenic factor1 (SF-1) regulation on porcine GnRHR2 gene expression and GnRH2 regulation mechanism in testosterone secretion through GnRHR2. Our study demonstrated that GnRHR2 transcription levels were high in porcine testis. Immunohistochemistry analyses showed that GnRHR2 immunoreactivity was strong in the Leydig cells in boar testes. Two SF-1 binding sites were predicted in GnRHR2 promoter and the second site (-159/-149) was considered to be important for GnRHR2 promoter activity through site-directed mutagenesis. The binding of SF-1 to GnRHR2 promoter was confirmed by electrophoretic mobility shift assays (EMSA) and chromatin immunoprecipitation (ChIP). Overexpression and knockdown experiments revealed that SF-1 could up-regulate porcine GnRHR2 expression. DNA methylation of GnRHR2 promoter CpG island also specifically regulated GnRHR2 expression. Meanwhile, our study also demonstrated GnRH2 treatment promoted the expression of SF-1 and steroidogenic acute regulatory protein (StAR), and that this treatment stimulated cAMP responsive element binding protein (CREB) phosphorylation, regulated the expression of GnRHR2, especially that of GnRHR2-X1, and promoted testosterone secretion in porcine Leydig cells. We speculated that testosterone secretion mediated by GnRH2 and GnRHR2 (mainly GnRHR2-X1) was regulated by phosphorylated CREB interacting with SF-1 to control StAR expression. Taken together, the present study indicates that SF-1 and GnRH2 are the essential regulatory factors for GnRHR2 expression. This study also explores the regulation mechanism of testosterone secretion mediated by GnRH2 and GnRHR2 in porcine Leydig cells.