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1.
Mol Immunol ; 126: 73-86, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32771671

RESUMO

Natural killer enhancing factor (NKEF) of peroxiredoxin family is an important innate immune molecule with having anti-oxidant activity. Although this gene has already been studied in a few fish species, it is yet to be identified and functionally characterised in Indian major carps. In the present study, the complete NKEF-B cDNA of rohu, Labeo rohita was cloned that encoded a putative protein of 197 amino acids. The phylogenetic study showed that L. rohita NKEF-B (LrNKEF-B) is closely related to NKEF-B of Cyprinus carpio and Danio rerio species. Tissue-specific expression of LrNKEF-B gene revealed the highest transcript level in the liver tissue. In the ontogeny study, the highest level of the expression was observed in milt and at 18 h post-development. The expression pattern of this gene was also studied in various pathogen models viz., Gram-negative bacteria (Aeromonas hydrophila), ectoparasite (Argulus siamensis) and a dsRNA viral analogue (poly I:C) in the liver and anterior kidney tissues of L. rohita juveniles. During A. hydrophila infection, the increase in expression of transcripts was observed at 3 h post-infection in both liver (15-fold) and anterior kidney (8-fold). In A. siamensis infection, the expression gradually increased up to 3 d post-infection in the anterior kidney, whereas in liver 3-fold up-regulation was noticed at 12 h post-infection. Similarly, during poly I:C stimulation, up-regulation of NKEF-B transcript was observed in anterior kidney from 1 h to 24 h post-stimulation and down-regulated afterwards whereas, the transcript level increased gradually from 6 h to 15 d post-stimulation in liver tissue. In vitro exposure to concanavalin, A and formalin-killed A. hydrophila upregulated NKEF-B gene expression in anterior kidney and peripheral blood leukocytes of L. rohita, however, down-regulated the same in the splenic leukocytes. A recombinant protein of LrNKEF-B (rLrNKEF-B) of 22 kDa was produced and it showed anti-oxidant activity by protecting supercoiled DNA and reducing insulin disulfide bonds. The minimum bactericidal concentration of this recombinant protein was found to be 4.54 µM against A. hydrophila and Staphylococcus aureus. Interestingly, rLrNKEF-B showed relative percent survival of 72.6 % in A. hydrophila challenged L. rohita, and the survival was found to be associated with a high level of expression of different cytokines, anti-oxidant genes and perforin in the rLrNKEF-B treated L. rohita. An indirect ELISA assay for estimation of NKEF was developed in L. rohita, and the concentrations of NKEF-B increased with time periods post A. hydrophila challenge viz., 0 h (42.56 ng/mL), 12 h (174 ng/mL) and 48 h (370 ng/mL) in rohu serum. Our results suggest a crucial role of LrNKEF-B in innate immunity against biotic stress and oxidative damage and also having antibacterial activity.


Assuntos
Carpas/imunologia , Doenças dos Peixes/imunologia , Proteínas de Peixes/imunologia , Imunidade Inata , Peroxirredoxinas/imunologia , Aeromonas hydrophila/imunologia , Animais , Arguloida/imunologia , Carpas/genética , Carpas/microbiologia , Carpas/parasitologia , Clonagem Molecular , Doenças dos Peixes/sangue , Doenças dos Peixes/microbiologia , Doenças dos Peixes/parasitologia , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Perfilação da Expressão Gênica , Rim Cefálico/enzimologia , Rim Cefálico/imunologia , Fígado/enzimologia , Fígado/imunologia , Estresse Oxidativo/imunologia , Peroxirredoxinas/genética , Filogenia , Poli I-C/imunologia , Espécies Reativas de Oxigênio/imunologia , Espécies Reativas de Oxigênio/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/metabolismo , Staphylococcus aureus/imunologia
2.
Arch Virol ; 165(7): 1599-1609, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32399788

RESUMO

The viral hemorrhage disease caused by grass carp reovirus (GCRV) is a serious contagious disease of grass carp that mainly infects fingerlings and yearlings. Epidemiological studies have shown that GCRV genotype II is currently the prominent genotype. However, little is known about the histopathological characteristics, virus distribution, and expression of immunity-related genes in grass carp infected by GCRV genotype II. In this study, we found that grass carp infected by GCRV genotype II lost appetite, swam alone, and rolled, and their fins, eyes, operculum, oral cavity, abdomen, intestine, and muscles showed pronounced punctate hemorrhage. Congestion, swelling, deformation, thinning of membranes, dilatation and darkened color of nucleoli, cathepsis, erythrocyte infiltration, and vacuole formation were observed in some infected tissues. A qRT-PCR test showed that the 11 genome segments of GCRV had similar expression patterns in different tissues. The S8 segment, with unknown function and no homologous sequences, had the highest expression level, while the most conserved segment, L2, had the lowest expression level. GCRV particles were distributed in different tissues, especially in the intestine. In the infected intestine, the expression of various receptors and adaptor molecules was modulated at different levels. Pro-inflammatory cytokine interleukin-1ß (IL-1ß) expression was 2160.9 times higher than that in the control group. The upregulation of immunity-related genes activated the antiviral immunity pathways. Therefore, the intestine might play a dual role in mediating GCRV infection and the antiviral immune response. This study provides detailed information about the pathogenicity of GCRV and expression of immunity-related genes, laying the foundation for further research on virus control and treatment.


Assuntos
Carpas/virologia , Doenças dos Peixes/genética , Proteínas de Peixes/genética , Infecções por Reoviridae/veterinária , Reoviridae/fisiologia , Animais , Carpas/genética , Carpas/imunologia , Doenças dos Peixes/imunologia , Doenças dos Peixes/patologia , Doenças dos Peixes/virologia , Proteínas de Peixes/imunologia , Genoma Viral , Genótipo , Interleucina-1beta/genética , Interleucina-1beta/imunologia , Reoviridae/genética , Infecções por Reoviridae/genética , Infecções por Reoviridae/imunologia , Infecções por Reoviridae/virologia
3.
J Fish Biol ; 96(6): 1399-1410, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32133636

RESUMO

Lgp2 (laboratory of genetics and physiology 2) is a cytosolic viral sensor of the RLR (retinoic acid-inducible gene 1 like receptor) family member without the caspase recruitment domain, having both inhibitory and stimulatory roles in RLR-signalling pathway. In India, Labeo rohita (rohu) is one of the leading and economically favoured freshwater fish species. Several immunological sentry proteins have been reported in this fish species, but no information is available on the RLR members. This study was aimed at cloning and characterization of full-length lgp2-cDNA (complementary DNA) in rohu and investigation of its expressional modulations following various pathogen-associated molecular pattern stimulations and bacterial infections. The full-length lgp2-cDNA sequence obtained through rapid amplification of cDNA ends-PCR consisted of 2299 nucleotides with an open reading frame of 2034 bp encoding 677 amino acids. In rohu-Lgp2, four conserved domains - a DEAD/DEAH box helicase domain, Pfam type-III restriction enzyme domain, helicase superfamily c-terminal domain and RIG-I C-terminal regulatory domain - have been detected. Within these domains, several important functional motifs, such as ATP-binding site, ATPase motif, RNA unwinding motif and RNA-binding sites, have also been identified. In healthy rohu, lgp2 gene was abundantly expressed in gill, liver, kidney, spleen and blood. In response to both in vitro and in vivo treatments using double-stranded RNA (poly I:C), lgp2 gene expression was significantly (P < 0.05) upregulated in all tested tissues and also in the LRG (Labeo rohita gill) cells. lgp2 gene expression significantly (P < 0.05) increased on stimulation of LRG cells using γ-d-glutamyl-meso-diaminopimelic acid and muramyl dipeptide. In vivo treatment using lipopolysaccharide and Aeromonas hydrophila-derived RNA resulted in both up- and down-regulation of lgp2 gene expression. Upon gram-positive and gram-negative bacterial infections, the expression of the lgp2 gene increased at different times in almost all the tested tissues. These integrated observations in rohu suggest that Lgp2 is an antiviral and antibacterial cytosolic receptor. SIGNIFICANCE STATEMENT: Lgp2, a cytosolic viral sensor of retinoic acid-inducible gene 1 like receptor family member, has been cloned in Labeo rohita. The complete sequence of rohu lgp2-complementary DNA consisted of 2299 nucleotides with an open reading frame of 2034 bp encoding 677 amino acids. It consisted of a DExDc, RES-III, HELICc, Pfam RIG-I_C-RD, ATP-binding site, ATPase motif, RNA unwinding motif and RNA-binding site. Upon bacterial infection, double-stranded RNA and various pathogen-associated molecular pattern stimulations, lgp2 gene expression significantly increased, indicating its role as an antiviral and antibacterial cytosolic receptor.


Assuntos
Carpas/genética , Carpas/imunologia , Proteínas de Peixes/genética , Infecções por Bactérias Gram-Negativas/imunologia , Aeromonas hydrophila/imunologia , Motivos de Aminoácidos , Sequência de Aminoácidos , Animais , DNA Complementar/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica
4.
Chemosphere ; 251: 126311, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32169710

RESUMO

The study provides cumulative data on the status of the two water bodies. The study designed revealed physicochemical properties (temperature, dissolved oxygen, pH, total dissolved solids and conductivity) to be in the desirable range, however, amongst the heavy metals excepting for Cd all were found to be higher than the permissible limits set by WHO and USEPA. It was observed that these elements cast their impact on bioindices (hepatosomatic index, condition factor, spleenosomatic index and kidney somatic index), renal marker enzyme (creatine kinase), hepatic marker enzymes (aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase), histology of immune organs (liver, spleen, head-kidney and thymus) and level of serum immunoglobulin (IgM). Further, expression levels of Metallothionein (MT) and Glutathione peroxidase (GPX) genes in immune-related tissues (liver, spleen, head-kidney, thymus and blood) observed indicates metal pollution and abiotic stresses. These alterations are reliable indicators of the cellular and humoral immune response in Cyprinus carpio.


Assuntos
Carpas/fisiologia , Fatores Imunológicos/toxicidade , Metais Pesados/toxicidade , Poluentes Químicos da Água/toxicidade , Animais , Biomarcadores/metabolismo , Carpas/genética , Carpas/imunologia , Expressão Gênica , Glutationa Peroxidase/genética , Glutationa Peroxidase/metabolismo , Rim Cefálico/efeitos dos fármacos , Fatores Imunológicos/metabolismo , Rim/metabolismo , Lagos , Fígado/metabolismo , Metalotioneína/genética , Metalotioneína/metabolismo , Metais Pesados/análise , Tanques , Baço/efeitos dos fármacos , Poluentes Químicos da Água/análise , Áreas Alagadas
5.
Appl Microbiol Biotechnol ; 104(4): 1633-1646, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31912200

RESUMO

Clonorchis sinensis (C. sinensis), an important fishborne zoonotic parasite threatening public health, is of major socioeconomic importance in epidemic areas. Effective strategies are still urgently expected to prevent against C. sinensis infection. In the present study, paramyosin of C. sinensis (CsPmy) was stably and abundantly expressed on the surface of Bacillus subtilis spores. The recombinant spores (B.s-CotC-CsPmy) were incorporated in the basal pellets diet in three different dosages (1 × 105, 1 × 108, 1 × 1011 CFU/g pellets) and orally administrated to grass carp (Ctenopharyngodon idella). The immune responses and intestinal microbiota in the treated grass carp were investigated. Results showed that specific anti-CsPmy IgM levels in sera, skin mucus, bile, and intestinal mucus, as well as mRNA levels of IgM and IgZ in the spleen and head kidney, were significantly increased in B.s-CotC-CsPmy-1011 group. Besides, transcripts levels of IL-8 and TNF-αin the spleen and head kidney were also significantly elevated than the control groups. Moreover, mRNA levels of tight junction proteins in the intestines of B.s-CotC-CsPmy-1011 group increased. Potential pathogenetic bacteria with lower abundance and higher abundances of candidate probiotics and bacteria associated with digestion in 1 × 1011 CFU/g B.s-CotC-CsPmy spores administrated fishes could be detected compared with control group. The amount of metacercaria in per gram fish flesh was statistically decreased in 1 × 1011 CFU/g B.s-CotC-CsPmy spores orally immunized group. Our work demonstrated that B. subtilis spores presenting CsPmy on the surface could be a promising effective, safe, and needle-free candidate vaccine against C. sinensis infection for grass carp.


Assuntos
Bacillus subtilis , Carpas/parasitologia , Clonorquíase/veterinária , Esporos Bacterianos , Tropomiosina/imunologia , Vacinas/administração & dosagem , Administração Oral , Ração Animal/microbiologia , Animais , Anticorpos Anti-Helmínticos/sangue , Carpas/imunologia , Cercárias/imunologia , Clonorquíase/imunologia , Clonorquíase/prevenção & controle , Clonorchis sinensis , Doenças dos Peixes/parasitologia , Doenças dos Peixes/prevenção & controle , Imunoglobulina M/imunologia , Intestinos/imunologia , Tropomiosina/genética , Vacinas/imunologia
6.
Fish Shellfish Immunol ; 98: 342-353, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31978531

RESUMO

Cyprinid Herpesvirus 3 (CyHV-3), also known as Koi Herpesvirus (KHV), causes Koi Herpesvirus Disease (KHVD) which leads to serious economic losses worldwide. To exploit DNA/subunit vaccine candidates, CyHV-3 ORF131 gene and cDNA was cloned and analyzed in the present study. Major B cell epitopes of deduced CyHV-3 pORF131 was also predicted. Then the complete CDS of CyHV-3 ORF131 was inserted into pEGFP-N1 vector and a modified pYD1/EBY100 system to construct the DNA and subunit vaccine, respectively. Subsequently, carp were immunized with homologous and heterologous prime-boost regimens relying on the constructed DNA and oral subunit vaccines. Then the protective immunity generated from different vaccines and regimens as well as the capacity of yeast (Saccharomyces cerevisiae) as an oral vaccine vehicle was evaluated. Our study confirmed that CyHV-3 ORF131 gene consisted of 2 introns and 3 exons encoding a 428 amino acids peptide. Further analysis indicated that four fragments of CyHV-3 pORF131 contained the major B cell epitopes (Cys20~Val140, Ser169~Tyr245, Thr258~Pro390, Phe414~Gln428), which could be linked and expressed in E. coli (BL21) as a truncated pORF131. The expression of full-length CyHV-3 pORF131 by pEGFP-N1 and yeast surface display was verified by In vitro assays before vaccination. Immunization of carp with CyHV-3 ORF131 DNA and subunit vaccines could evoke the activation of immune-related genes such as CXCa, CXCR1, IL-1ß, TNF-α, INF-a1, Mx-1, IgM, IgT1 and production of specific serum IgM measured by ELISA. RPS (relative percent of survival) ranging from 53.33% to 66.67% was acquired post challenge test. Moreover, flow cytometry analysis illustrated the delivery of surface-displayed CyHV-3 pORF131 to midgut after oral gavage. Thus, our findings suggest that CyHV-3 ORF131 can serve as DNA/subunit vaccines candidate and the yeast as an ideal oral vaccine vehicle.


Assuntos
Carpas , Doenças dos Peixes/prevenção & controle , Infecções por Herpesviridae/veterinária , Herpesviridae/imunologia , Vacinas contra Herpesvirus/imunologia , Fases de Leitura Aberta/imunologia , Vacinação/veterinária , Administração Oral , Animais , Anticorpos Antivirais/sangue , Carpas/imunologia , Carpas/virologia , Técnicas de Visualização da Superfície Celular , Epitopos de Linfócito B , Escherichia coli/genética , Doenças dos Peixes/virologia , Regulação da Expressão Gênica/imunologia , Infecções por Herpesviridae/prevenção & controle , Vacinas contra Herpesvirus/administração & dosagem , Esquemas de Imunização , Fases de Leitura Aberta/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/metabolismo , Saccharomyces cerevisiae/genética , Análise de Sobrevida , Vacinação/métodos , Vacinas de DNA/administração & dosagem , Vacinas de DNA/imunologia , Vacinas de Subunidades/administração & dosagem , Vacinas de Subunidades/imunologia
7.
Fish Shellfish Immunol ; 98: 324-333, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31981775

RESUMO

Microbial exopolysaccharides (EPS) from Lactococcus have been found to have an important role in the probiotic activity of this bacterium; however, the immunomodulatory and antioxidant activities have not been fully explored in aquaculture. In the present study, we investigated EPS-2 from Lactococcus lactis Z-2, isolated from healthy common carp, for its immunomodulatory and antioxidant effects and disease resistance against Aeromonas hydrophila in Cyprinus carpio L. We found that the molecular weight of EPS-2 was 18.65 KDa. The monosaccharide composition of this polymer was rhamnose, xylose, mannose, glucose, and galactose at a molar percentage of 13.3%, 14.1%, 18.5%, 27.4%, and 26.7%, respectively. EPS-2 treatment could modulate the immune responses in vitro and in vivo. In vitro tests showed that EPS-2 could significantly enhance the proliferation and phagocytosis activities (P < 0.05) as well as induce the production of nitic oxide (NO), pro-inflammatory cytokines (TNF-α, IL-1ß, IL-6), and anti-inflammatory cytokines (IL-10, TGF-ß) (P < 0.05) in head kidney cells. When the fish were gavaged with three different concentrations of EPS-2 (250, 500, 1000 µg/mL) for 7 days and infected with A. hydrophila, different expression patterns of the NO, cytokines, lysozyme (LZM), and alkaline phosphatase (AKP) in the serum and of antioxidants (T-AOC, SOD, CAT, GSH, GSH-Px and MDA) in hepatopancreas were observed. Before infection with A. hydrophila, EPS-2 supplementation significantly up-regulated the NO production, protein levels of pro-inflammatory cytokines (TNF-α, IL-1ß, IL-6), LZM and AKP activities, and levels of antioxidant molecules compared to those in the negative (G1) group (P < 0.05), whereas levels of NO and pro-inflammatory cytokines and LZM and AKP activities were significantly lower than those in the positive (G2) group after infection (P < 0.05). However, whether infected or not, the expression levels of anti-inflammatory cytokines (IL-10, TGF-ß) were significantly increased in the EPS-2 treatment groups (P < 0.05). These results indicate that EPS-2 has immunomodulatory and antioxidant effects on common carp, both in vitro and/or in vivo, and can be applied as a common carp feed supplement to enhance fish immunity and disease resistance against A. hydrophila.


Assuntos
Aeromonas hydrophila/fisiologia , Antioxidantes/metabolismo , Carpas/imunologia , Resistência à Doença/efeitos dos fármacos , Imunidade Inata/efeitos dos fármacos , Lactococcus lactis/química , Polissacarídeos Bacterianos/farmacologia , Animais , Carpas/microbiologia , Proliferação de Células , Citocinas/metabolismo , Suplementos Nutricionais , Rim Cefálico/citologia , Rim Cefálico/efeitos dos fármacos , Rim Cefálico/imunologia , Óxidos de Nitrogênio/metabolismo , Fagocitose , Probióticos/farmacologia
8.
Fish Shellfish Immunol ; 98: 25-33, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31904539

RESUMO

Mannose-binding lectin (MBL) is a crucial pattern recognition receptor in the host innate immune system. Previously, we reported the biological function of Ctenopharyngodon idella MBL (CiMBL) in initiating the lectin pathway of the complement system. In the present study, we further explored its biological function including the agglutinating ability, binding capacity and protective role in vitro and in vivo. After Aeromonas hydrophila infection, western blot analysis revealed that the CiMBL were fluctuated and expressed in the serum and major immune-related tissues. The result of quantitative PCR (qPCR) showed that the recombinant CiMBL (rCiMBL) significantly inhibited the mRNA expression of interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α) in liver, spleen and hepatic cells. Due to rCiMBL bound to d-mannose, d-galactose, d-glucose, N-acetyl-d-glucosamine (GlcNAc), lipopolysaccharide (LPS), peptidoglycan (PGN) and Agar in the presence of Ca2+, herein gram-positive (Staphylococcus aureus and Micrococcus luteus) and gram-negative (A. hydrophila and Vibrio anguillarum) bacteria were agglutinated by rCiMBL in a Ca2+-dependent manner. More importantly, rCiMBL enhanced the survival rate of grass carp following bacterial infection. Overall, the results provide an evidence that CiMBL can protect grass carp against A. hydrophila infection in aquaculture.


Assuntos
Aglutinação , Carpas/imunologia , Doenças dos Peixes/imunologia , Lectina de Ligação a Manose/imunologia , Monossacarídeos/metabolismo , Polissacarídeos/metabolismo , Aeromonas hydrophila/fisiologia , Animais , Carpas/metabolismo , Proteínas de Peixes/imunologia , Proteínas de Peixes/farmacologia , Bactérias Gram-Negativas/fisiologia , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Bactérias Gram-Positivas/fisiologia , Lectina de Ligação a Manose/farmacologia , Substâncias Protetoras/farmacologia
9.
Vet Immunol Immunopathol ; 221: 110009, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31945652

RESUMO

A 14-day experiment was conducted to explore the pathological process and immune response of soybean meal (SBM) induced enteritis (SBMIE) in grass carp (Ctenopharyngodon idellus). The complete replacement of dietary fish meal (FM) with SBM resulted in a remarkable reduction in final body weight, weight gain ratio, and feed conversion efficiency (p < 0.05). The typical histopathological changes of SBMIE appeared starting at day 4, and progressively increased in severity until day 8, then gradually subsided after day 11. The course of SBMIE could be divided into incubation period (days 1-2), prodromal period (days 3-6), symptomatic period (days 7-10), and convalescent period (days 11-14). Transcription levels of pro-inflammatory cytokines, including IL-1ß, TNF-α, IL-6, IL-8, IL-17A/F1 and IFN-γ2, were up-regulated during the prodromal period, and then down-regulated during the convalescent period. Transcript levels of anti-inflammatory cytokines (IL-10 and TGFß1) and their receptors (IL-10R1 and TßRII), were up-regulated during the prodromal and convalescent periods. Transcript levels of MHCIIß, Igµ, Igτ, TCRδ, TCRß, CD4, and CD8α were altered in SBMIE. Furthermore, expression levels of T-bet, IFN-γ2, RORγ2 and IL-17A/F1 were significantly increased in the initiation of enteritis, whereas the transcript levels of Foxp3 and IL-2/15Ra were significantly up-regulated in the repair of enteritis. In conclusion, grass carp SBMIE is regulated by the adjustment of SBM-based diet intake, and the changes of the above-mentioned genes expression suggest that these genes may be involved in SBMIE.


Assuntos
Ração Animal/análise , Carpas/imunologia , Citocinas/imunologia , Enterite/veterinária , Doenças dos Peixes/imunologia , Trato Gastrointestinal/imunologia , Soja/efeitos adversos , Animais , Carpas/metabolismo , Citocinas/genética , Suplementos Nutricionais , Enterite/induzido quimicamente , Enterite/imunologia , Doenças dos Peixes/induzido quimicamente , Trato Gastrointestinal/patologia , Inflamação/genética , Soja/química
10.
Artigo em Inglês | MEDLINE | ID: mdl-31958508

RESUMO

Bisphenol A (BPA) is a well-known phenolic environmental estrogen, widely distributed in the aquatic environment, which poses a toxic risk to the health of aquatic organisms. This study aimed to assess the effect of BPA on common carp gills by analyzing oxidative stress, ion equilibrium and immune response. Fish were exposed to five concentrations of BPA (0, 0.01, 0.1, 0.5, and 2 mg/L) for 30 days. Then gills were collected to assay biochemical parameters and gene expression. The results showed that BPA could decrease the levels of total antioxidant capacity (T-AOC), catalase (CAT), glutathione (GSH) and glutathione S-transferase (GST) and increase the levels of superoxide dismutase (SOD), malondialdehyde (MDA) and 8-hydroxy-2 deoxyguanosine (8-OHdG). The gene expression showed that BPA (2 mg/L) could affect the nuclear erythroid 2-related factor 2 (nrf2) signaling pathway, upregulate the gene expression of nrf2 and heme oxygenase 1 (ho-1). Meanwhile, BPA was found to change the activity of Na+/K+ ATPase, and increased the concentrations of Na+ and Ca2+ in gills of common carp. Also, high BPA concentration (0.5 or 2 mg/L) exposure increased the activity of alkaline phosphatase (AKP), blocked mRNA level of lysozyme-c (c-lyz), activated Toll-like receptors (TLRs) signaling pathway, enhanced the mRNA levels of toll-like receptor 2 (tlr2), receptor 4 (tlr4), myeloid differentiation factor 88 (myd88), interferon regulatory factor 3 (irf3), interleukin 1ß (il-1ß), interleukin 6 (il-6) and interleukin 10 (il-10). Overall, these results suggested that high BPA could induce oxidative damage, ion imbalance, immunosuppression and inflammatory response in gills of common carp.


Assuntos
Compostos Benzidrílicos/toxicidade , Carpas , Proteínas de Peixes , Brânquias/metabolismo , Fenóis/toxicidade , Poluentes Químicos da Água/toxicidade , Animais , Carpas/imunologia , Carpas/metabolismo , Proteínas de Peixes/imunologia , Proteínas de Peixes/metabolismo , Transporte de Íons , Estresse Oxidativo
11.
Fish Shellfish Immunol ; 96: 190-200, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31765792

RESUMO

Complement is a complex component of innate immune system, playing an important role in defense against pathogens and host homeostasis. The complement system has been comprehensively studied in mammals, however less is known about complement in teleost, especially in tetraploid common carp (Cyprinus carpio). In this study, a total of 110 complement genes were identified and characterized in common carp, which include almost all the homologs of mammalian complement genes. These genes were classified into three pathways (alternative pathways, lectin pathways and classical pathways), similar to those in mammals. Phylogenetic and selection pressure analysis showed that the complement genes were evolving-constrained and the function was conserved. Most of the complement genes were highly expressed in spleen, liver, brain and skin among the tested 12 health tissues of common carp. After Aeromonas hydrophila infection in the common carp, many members of complement genes were activated to bring about an immune response and expressed to against any pathogenic encroachment. Gene expression divergences which were found between two homoeologous genes suggested the functional divergences of the homoeologous genes after the 4R WGD event, revealing the evolutionary fate of the tetraploid common carp after the recent WGD.


Assuntos
Carpas/genética , Proteínas do Sistema Complemento/genética , Doenças dos Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Genoma/imunologia , Imunidade Inata/genética , Aeromonas hydrophila/fisiologia , Animais , Carpas/imunologia , Proteínas do Sistema Complemento/imunologia , Evolução Molecular , Proteínas de Peixes/química , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Perfilação da Expressão Gênica/veterinária , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Filogenia , RNA Mensageiro/genética
12.
Fish Shellfish Immunol ; 96: 26-31, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31794841

RESUMO

Di-(2-ethylhexyl) phthalate (DEHP), a common pollutant in the water environment, has been reported to be associated with immune functions, especially aquatic organisms. However, whether DEHP exposure causes neutrophils toxicity in common carp is still unclear. To investigate the toxic effect of DEHP on immune functions, common carp neutrophils were exposed to DEHP (40 µmol/L and 200 µmol/L) for 2 h. The common carp neutrophils exposed to DEHP showed a decrease in neutrophil phagocytosis rate compared with control group. DEHP exposure induced a significant decrease in mRNA expression levels of inflammatory cytokines-related genes (Interleukin-6, Interleukin-8, transforming growth factor, tumor necrosis factor (TNF)-α, TNF-R1, TNF-T1, Interferon (IFN)-2a, IFN-g2b, IFN-g1) in common carp neutrophils, while the expression levels of IL-1ß and IL-10 were increased compared with control group (P < 0.05). Furthermore, the detection of cytochrome P450 enzyme related genes showed that the mRNA expression levels of CYP (cytochrome P450 proteins)-1A, CYP-1B1, CYP-C1, CYP-2K were significantly decreased, and the mRNA expression level of CYP-3A was significantly reduced (P < 0.05). The results indicated that DEHP could affect the phagocytic ability of neutrophils by regulating the expression of inflammatory cytokines and disrupting cytochrome P450 homeostasis, which caused the immunosuppression in common carp.


Assuntos
Carpas/imunologia , Sistema Enzimático do Citocromo P-450/imunologia , Dietilexilftalato/efeitos adversos , Proteínas de Peixes/metabolismo , Tolerância Imunológica/efeitos dos fármacos , Neutrófilos/imunologia , Poluentes Químicos da Água/efeitos adversos , Animais , Carpas/metabolismo , Homeostase/imunologia , Neutrófilos/efeitos dos fármacos , Plastificantes/efeitos adversos
13.
Dev Comp Immunol ; 102: 103475, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31437525

RESUMO

In mammals, interferon regulatory factor 5 (IRF5) can be activated by tumor necrosis factor receptor-associated factor 6 (TRAF6). Upon activation, IRF5 translocates into the nucleus, where it binds to IFN promoter and up-regulates IFN expression. However, there are few reports on the molecular mechanism by which TRAF6 up-regulates IFN expression in fish. In this study, we explored how Grass carp (Ctenopharyngodon idellus) TRAF6 initiated innate immunity by activating IRF5. We found that CiTRAF6, CiIRF5 and CiIFN1 were all significantly up-regulated in LPS-stimulated CIK cells and the expression of CiTRAF6 was earlier than the expressions of CiIRF5 and CiIFN1. These findings suggested that CiIFN1 expression might be induced by CiTRAF6 in fish. CiIFN1 expression, CiIFN1 promoter activity and CO cells viability were all significantly up-regulated in the overexpression experiments, but they were significantly down-regulated in the gene silencing experiments. This indicated that CiTRAF6, along with CiIRF5, regulated CiIFN1 expression. The localization analysis found that both CiTRAF6 and CiIRF5 located in the cytoplasm. Following LPS stimulation, CiIRF5 was observed to translocate to the nucleus. GST-pull down and co-IP experiments revealed that CiTRAF6 interacted with CiIRF5. The colocalization analysis also showed that CiTRAF6 bound with CiIRF5 in the cytoplasm. Overexpression of CiTRAF6 increased the endogenous CiIRF5, promoted its ubiquitination and nuclear translocation. In conclusion, CiTRAF6 bound to CiIRF5 in the cytoplasm, and then activated CiIRF5, resulting in up-regulating the expression of CiIFN1.


Assuntos
Carpas/imunologia , Proteínas de Peixes/metabolismo , Fatores Reguladores de Interferon/metabolismo , Interferon Tipo I/metabolismo , Fator 6 Associado a Receptor de TNF/metabolismo , Animais , Núcleo Celular/metabolismo , Sobrevivência Celular , Células Cultivadas , Proteínas de Peixes/genética , Imunidade Inata , Fatores Reguladores de Interferon/genética , Interferon Tipo I/genética , Lipopolissacarídeos/farmacologia , Ligação Proteica , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/imunologia , Fator 6 Associado a Receptor de TNF/genética , Ativação Transcricional/efeitos dos fármacos , Ubiquitinação/efeitos dos fármacos
14.
Dev Comp Immunol ; 102: 103488, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31476324

RESUMO

This study aimed to evaluate the influence of dietary pure linseed oil or sesame oil or a mixture on innate immune competence and eicosanoid metabolism in common carp (Cyprinus carpio). Carp of 100.4 ±â€¯4.7 g were fed to satiation twice daily for 6 weeks with four diets prepared from three lipid sources (CLO; LO; SO; SLO). On day 42, plasma was sampled for immune parameter analyses, and kidney and liver tissues were dissected for gene expression analysis. On day 45, HKL and PBMCs from remaining fish were isolated and exposed to E. coli LPS at a dose of 10 µg/mL for 24 h. Results show that the SLO diet enhanced feed utilisation (P = 0.01), while no negative effects on growth or survival were observed in plant oil-fed fish compared to those fed a fish-oil based diet. Plant oil diets did not alter lysozyme and peroxidase activities or gene expression levels. Moreover, the diets did not affect the expression levels of some genes involved in eicosanoid metabolism processes (pla, pge2, lox5). Lys expression in HKL in vitro following exposure to LPS was up-regulated in LO-fed fish, while expression levels of pge2 were higher in SLO fish than in other groups (P < 0.05). The highest value for peroxidase activity in HKL exposed to LPS was found in the SLO-fed group (P < 0.05). In conclusion, our results indicate that dietary plant oils did not induce any negative effects on fish growth, survival, and immune competence status. Moreover, a dietary combination of SO and LO improved the feed utilisation efficiency and seemed more effective in inducing a better immunomodulatory response to LPS through a more active eicosanoid metabolism process.


Assuntos
Carpas , Eicosanoides/metabolismo , Imunidade Inata/fisiologia , Óleo de Semente do Linho/metabolismo , Óleo de Gergelim/metabolismo , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Carpas/imunologia , Carpas/metabolismo , Células Cultivadas , Ácidos Graxos/metabolismo , Rim Cefálico/citologia , Rim Cefálico/imunologia , Imunidade Inata/genética , Leucócitos/metabolismo , Óleo de Semente do Linho/análise , Metabolismo dos Lipídeos/genética , Lipopolissacarídeos/farmacologia , Muramidase/metabolismo , Peroxidase/metabolismo , Óleo de Gergelim/análise
15.
Fish Shellfish Immunol ; 97: 540-553, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31881329

RESUMO

This study investigated the effects of dietary curcumin on growth performance, non-specific immunity, antioxidant capacity and related genes expression of NF-κB and Nrf2 signaling pathways in grass carp (Ctenopharyngodon idella). A total of 525 juvenile grass carps with mean initial body weight of (5.30 ± 0.10) g were randomly distributed into five groups with three replicates each, fed five diets containing graded levels of curcumin (0, 196.11, 393.67, 591.46 and 788.52 mg/kg diet) for 60 days. After feeding trial, fifteen fish per tank were challenged with Aeromonas hydrophila and the mortalities were recorded for 7 days. The results showed that optimal dietary curcumin (393.67 mg/kg diet) improved the weight gain (WG) and specific growth rate (SGR) of juvenile grass carp, reduced feed conversion ratio (FCR) and the mortalities after challenge (P < 0.05). Moreover, optimal dietary curcumin increased the activities of lysozyme (LYZ) and acid phosphatase (ACP), and complement 3 (C3) and C4 levels, decreased alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in serum of grass carp after injection with A. hydrophila (P < 0.05). Meanwhile, optimal dietary curcumin up-regulated the mRNA levels of LYZ, C3 and antimicrobial peptides [hepcidin, liver-expressed antimicrobial peptide-2 (LEAP-2), ß-defensin], and anti-inflammatory cytokines of interleukin-10 (IL-10) and transforming growth factor ß1 (TGF-ß1), and inhibitor of κBα (IκBα), whereas down-regulated pro-inflammatory cytokines of tumor necrosis factor-α (TNF-α), IL-1ß, IL-6 and IL-8, and nuclear factor kappa B p65 (NF-κB p65), IκB kinases (IKKα, IKKß and IKKγ) mRNA levels in the liver and blood of grass carp after injection with A. hydrophila (P < 0.05). In addition, optimal dietary curcumin increased the reduced glutathione (GSH) content and activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione S-transferase (GST) and glutathione reductase (GR), reduced reactive oxygen species (ROS) and malondialdehyde (MDA) levels in the liver of grass carp after injection with A. hydrophila (P < 0.05). Meanwhile, optimal dietary curcumin up-regulated the mRNA levels of these antioxidant enzymes and nuclear factor erythroid 2-related factor 2 (Nrf2), whereas down-regulated Kelch-like ECH-associated protein (Keap) 1a and Keap 1b mRNA levels (P < 0.05) in the liver and blood of grass carp after injection with A. hydrophila. Thus, optimal dietary curcumin supplementation could promote growth of juvenile grass carp, reduce FCR, and enhance disease resistance, innate immunity and antioxidant capacity of fish, attenuating inflammatory response. However, dietary excessive curcumin had negative effect on fish. Based on second-order regression analysis between dietary curcumin contents and weight gain, the optimum requirement of dietary curcumin in juvenile grass carp was determined to be 438.20 mg/kg diet.


Assuntos
Carpas/crescimento & desenvolvimento , Carpas/imunologia , Curcumina/farmacologia , Suplementos Nutricionais/análise , Proteínas de Peixes/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Transdução de Sinais , Aeromonas hydrophila , Ração Animal/análise , Animais , Antioxidantes/metabolismo , Carpas/microbiologia , Citocinas/imunologia , Resistência à Doença , Doenças dos Peixes/imunologia , Doenças dos Peixes/microbiologia , Doenças dos Peixes/prevenção & controle , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/prevenção & controle , Imunidade Inata , Fator 2 Relacionado a NF-E2/imunologia , NF-kappa B/imunologia
16.
Fish Shellfish Immunol ; 97: 500-508, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31883471

RESUMO

Interleukin (IL)-2 belongs to the four-helix bundle cytokine family and plays key roles in growth, survival, activation-induced cell death and differentiation of the immune cells. In cyprinid fish, only common carp interleukin-2 (il2) has been cloned because of relatively low sequence homology between carp Il-2 and its homologs in other fish species. In the present study, the coding sequence of grass carp Il-2 (gcIl-2) was cloned and its identity was verified via bioinformatic analysis. Tissue distribution study showed that grass carp il2 (gcil2) mRNA was expressed in thymus, head kidney and gill with relatively high levels. Recombinant gcIl-2 (rgcIl-2) protein was subsequently prepared by using a prokaryotic expression system followed by a refolding method. The purified rgcIl-2 displayed an ability to stimulate the cell proliferation along with an increased mRNA expression of cd4l but not cd8a, igm or mcsfr in grass carp head kidney leukocytes (HKLs), suggesting the possible involvement of gcIl-2 in T helper (Th) cell proliferation. In the same cell model, rgcIl-2 significantly enhanced mRNA expression of some cytotoxic molecules including perforin-like protein 2, granzyme B-like and Fas ligand, indicating the modulation of cytotoxic cells by gcIl-2 in grass carp HKLs. Besides, gene expression of regulatory T (Treg) cell- and Th1/2 cell-related cytokines or transcription factors was detected in grass carp HKLs treated by rgcIl-2. Results showed that rgcIL2 treatment increased the mRNA expression of foxp3, cd25l, ifng2, il12p35, tbet, tnfa, il2, il4/13a, il4/13b and gata3l in HKLs, implying the regulatory roles of Il-2 in the expression of these immune genes and its possible involvement in differentiation of Treg and Th1/2 cells. These observations together with the related studies in other fishes suggest the existence of cytotoxic cells, Treg and Th1/2 subpopulations in fish species and the functional roles of Il-2 in these cells.


Assuntos
Carpas/imunologia , Rim Cefálico/citologia , Interleucina-2/imunologia , Leucócitos/imunologia , Animais , Biologia Computacional , Citocinas/imunologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Expressão Gênica , Rim Cefálico/imunologia , Interleucina-2/genética , RNA Mensageiro/imunologia , Transdução de Sinais
17.
Fish Shellfish Immunol ; 97: 344-350, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31846776

RESUMO

To study the effect of dietary supplementation of Bacillus licheniformis FA6 on the growth, survival and intestinal health of grass carp, we assessed the antioxidant capacity, intestinal barrier, expression levels of immune genes, and the resistance to Aeromonas hydrophila AH-1 infection. Experimental setup comprised three groups (90 specimens each; average initial weight = 16.5 g): the control group was fed the basal diet without B. licheniformis, the low-dose (LD) group was supplemented with B. licheniformis at the concentration of 1 × 105 cfu/g, and the high-dose (HD) group with 1 × 106 cfu/g. After 56 days of growth trial, the challenge test with A. hydrophila AH-1 was conducted for 14 days. The results revealed that the grass carp in LD group and HD group had significantly (p < 0.05) improved percent weight gain (PWG) and specific growth rate (SGR) parameters. Additionally, the antioxidant status was improved, which included increased superoxide dismutase (SOD) activity and decreased malondialdehyde (MDA) levels in the serum, and upregulated mRNA levels of antioxidant enzymes MnSOD and catalase (CAT) in the intestine. Meanwhile, B. licheniformis FA6 supplementation groups exhibited a decreased mRNA expression of proinflammatory cytokines (such as IL-1ß and TNF-α) and increased the expression of anti-inflammatory cytokine IL-10. Histological (villi length was increased) and gene expression (qPCR: upregulated ZO-1, occludin, and claudin-c) analyses suggested improved functioning of the intestinal barrier. Post-challenge mortality rates in LD and HD groups were significantly lower (56.6% and 70% respectively) than in the control group (100%). Overall, these results indicated that dietary supplementation of B. licheniformis FA6 can improve growth, antioxidant capacity, intestinal barrier functions and disease resistance of grass carp.


Assuntos
Antioxidantes/metabolismo , Bacillus licheniformis/química , Carpas/imunologia , Intestinos/imunologia , Probióticos/farmacologia , Ração Animal/análise , Animais , Carpas/crescimento & desenvolvimento , Carpas/metabolismo , Dieta/veterinária , Resistência à Doença/fisiologia , Relação Dose-Resposta a Droga , Doenças dos Peixes/imunologia , Probióticos/administração & dosagem , Distribuição Aleatória
18.
Fish Shellfish Immunol ; 97: 351-358, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31874297

RESUMO

This study was conducted to investigate the effect of dietary Scenedesmus ovalternus on the growth and disease resistance of gibel carp (Carassius gibelio) during overwintering. Gibel carp (initial body weight: 90.39 ± 0.33 g) were fed with diets containing 0% or 4% Scenedesmus ovalternus (DS0 and DS4) for 4 weeks during the early overwintering period, and then all fish were left unfed during the late overwintering period. A bacterial challenge test using Aeromonas hydrophila was subsequently conducted. The 4% Scenedesmus ovalternus diet had no effect on the growth of gibel carp (P > 0.05), but did improve the survival rate after the challenge (P ≤ 0.05). In the DS0 group, the bacterial challenge decreased the contents of complement 3 (C3), immunoglobulin M (IgM), interleukin 2 (IL2) and tumor necrosis factor α (TNFα) in fish (P < 0.05); in the DS4 group, the challenge increased total antioxidant capacity (T-AOC) and myeloperoxidase (MPO) activity but decreased IL2 and TNFα contents (P < 0.05). The activities of MPO and contents of C3, IgM and TNFα were higher in the DS4 group than that fed the DS0 diet after bacterial challenge (P < 0.05). Compared to pre challenge, the expression levels of toll like receptor 2 (TLR2), toll like receptor 3 (TLR3), toll like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), Toll/IL-1 receptor domain-containing adaptor protein (TIRAP), TIR-domain-containing adapter-inducing interferon ß (TRIF), nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor α (IκBα), transforming growth factor ß (TGFß), interleukin 1ß (IL1ß), tumor necrosis factor α1 (TNFα1) and interleukin 10 (IL10) in the head kidney of gibel carp were induced after challenge (P < 0.05). Gibel carp fed the DS4 diet showed lower expression of TGFß in head kidney before the challenge and lower expression of TLR2, TLR3, TLR4, TIRAP, TRIF, IκBα, TNFα1, IL10 and TGFß after the challenge than that fed the DS0 diet (P < 0.05). Overall, Scenedesmus ovalternus supplement enhanced the resistances of gibel carp against A. hydrophila after overwintering via the TLR signaling pathway.


Assuntos
Carpas/genética , Carpas/imunologia , Resistência à Doença/genética , Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Scenedesmus/química , Receptores Toll-Like/genética , Aeromonas hydrophila/fisiologia , Ração Animal/análise , Animais , Dieta/veterinária , Suplementos Nutricionais/análise , Doenças dos Peixes/genética , Proteínas de Peixes/metabolismo , Infecções por Bactérias Gram-Negativas/genética , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Distribuição Aleatória , Análise de Sequência de DNA/veterinária , Transdução de Sinais/genética , Receptores Toll-Like/metabolismo
19.
Mol Immunol ; 118: 79-90, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31855810

RESUMO

The p38 mitogen-activated protein kinases (MAPKs) are essential cytoplasmic signal molecules of innate immune pathways that play a vital role in host immune defense responses to pathogenic challenges. In this study, two fish p38 genes (Cip38α and Cip38ß) were characterized for the first time from the grass carp Ctenopharyngodon idella. Similar to other reported p38MAPKs, both Cip38α and Cip38ß contained a conserved phosphorylation motif (Thr-Gly-Tyr, TGY) and a substrate binding site (Ala-Thr-Arg-Trp, ATRW) in the serine/threonine protein kinase (S_TKc) domain. Expression profile analysis showed that Cip38α and Cip38ß mRNAs were broadly expressed in all of the examined tissues and developmental stages of C. idella. In addition, in vivo injection experiments directly revealed that Cip38α and Cip38ß showed strong responsiveness to Aeromonas hydrophila and muramyl dipeptide (MDP) challenges, and their expression levels were significantly upregulated in the intestine of grass carp. Additionally, the MDP-induced expression levels of intestinal inflammatory cytokines (TNF-α and IL-15) and an antimicrobial peptide (ß-defensin) were significantly inhibited by the p38MAPK-specific inhibitor SB203580. Moreover, the nutritional dipeptide carnosine and Ala-Gln were found to significantly suppress the bacterial MDP-induced expression of p38MAPK pathway genes and inflammatory cytokines in the intestine of grass carp. Finally, overexpression analysis demonstrated that Cip38α and Cip38ß could act as efficient activators in the regulation of AP-1 signaling pathways through interaction with CiMKK6. Altogether, this study provided experimental evidence of the presence of a functional p38 pathway in grass carp, which revealed its involvement in the intestinal immune response to bacterial challenges in bony fish.


Assuntos
Acetilmuramil-Alanil-Isoglutamina/imunologia , Carpas/imunologia , Carpas/microbiologia , Imunidade Inata/imunologia , Intestinos/imunologia , Proteínas Quinases p38 Ativadas por Mitógeno/imunologia , Aeromonas hydrophila/imunologia , Animais , Citocinas/imunologia , Doenças dos Peixes/imunologia , Doenças dos Peixes/microbiologia , Proteínas de Peixes/imunologia , Células HEK293 , Humanos , Inflamação/imunologia , Inflamação/microbiologia , Intestinos/microbiologia , NF-kappa B/imunologia , Transdução de Sinais/imunologia
20.
Fish Shellfish Immunol ; 98: 34-44, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31883472

RESUMO

Ctenopharyngodon idella growth arrest and DNA damage-inducible 45 ab (CiGadd45ab) is a subtype of the Gadd45a gene of the Gadd45 family in grass carp. There is increasing evidence that microRNAs (miRNAs) are involved in the regulation of inflammatory and apoptotic responses. However, little is known about the regulatory effects of miRNAs on CiGadd45ab expression. In the present study, CiGadd45ab was identified as a target gene of miR-23a-3p and miR-23a-5p, based on miRNA expression profiling and a dual-luciferase reporter assay. In addition, miR-23a-3p and miR-23a-5p were both confirmed to be involved in the inflammatory response following infection with Aeromonas hydrophila by targeting CiGadd45ab. Transfection with miR-23a-3p and miR-23a-5p mimics and inhibitor altered proinflammatory gene expression and apoptosis rate, thereby suggesting that miRNAs regulate immune response and anti-apoptosis by targeting CiGadd45ab in grass carp. Our results provide a theoretical basis for exploring the molecular mechanisms by which miR-23a-3p and miR-23a-5p target CiGadd45ab to regulate inflammation and apoptosis against bacterial infection in grass carp.


Assuntos
Apoptose/imunologia , Carpas/genética , Carpas/imunologia , Doenças dos Peixes/imunologia , Inflamação/veterinária , Peptídeos e Proteínas de Sinalização Intracelular/genética , MicroRNAs/genética , Aeromonas hydrophila/fisiologia , Animais , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Inflamação/imunologia , Peptídeos e Proteínas de Sinalização Intracelular/imunologia , MicroRNAs/imunologia
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