Molecular cloning, characterization and expression analysis of PPAR gamma in the orange-spotted grouper (Epinephelus coioides) after the Vibrio alginolyticus challenge

Luo, Sheng‐Wei; Huang, Youhua; Xie, Fu-Xing; Huang, Xiaohong; Liu, Yuan; Wang, Weina; Qin, Qiwei

doi:10.1016/j.fsi.2015.01.003

Cited by 41 publications

(9 citation statements)

References 66 publications

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“…Similar observations were also described for fish. For example, overexpression of PPARγ in spleen cells of the orange-spotted grouper decreased the expression of proinflammatory mediators, whereas blocking of PPARγ induced an increase in inflammation [89]. Interestingly, in human macrophages, as with dexamethasone, GW9662 shifted the phenotype of macrophages from M2a to M2c, characterized by an inhibited production of TNF-α and IL-10 [90].…”

Section: Discussionmentioning

confidence: 99%

“…To verify involvement of macrophage-derived cortisol in macrophage polarization, cells were pretreated for 1 h with an inhibitor of cortisol synthesis/conversion-metyrapone (Sigma-Aldrich, St. Louis, MO, USA; 1 µM, 10 µM), while, to study role of GRs and PPARγ in this process, cells were pretreated with GR antagonist-RU-486 (Sigma-Aldrich, St. Louis, MO; 1 µM [93]) or selective PPARγ antagonist-GW9662 (Sigma-Aldrich, St. Louis, MO; 1 µM, 10 µM [89]). All inhibitors were dissolved in dimethyl sulfoxide (DMSO) (Bioshop, Burlington, ON, Canada); therefore, control cells were pretreated with vehicle-0.1%, DMSO.…”

Section: Inhibition Of Cortisol Conversion and Blocking Of Gr And Pparγmentioning

confidence: 99%

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Cortisol Metabolism in Carp Macrophages: A Role for Macrophage-Derived Cortisol in M1/M2 Polarization

Maciuszek

Kłak

Rydz

et al. 2020

IJMS

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Macrophages are crucial not only for initiation of inflammation and pathogen eradication (classically polarized M1 macrophages), but also for inflammation inhibition and tissue regeneration (alternatively polarized M2 macrophages). Their polarization toward the M1 population occurs under the influence of interferon-γ + lipopolysaccharide (IFN-γ + LPS), while alternatively polarized M2 macrophages evolve upon, e.g., interlukin 4 (IL-4) or cortisol stimulation. This in vitro study focused on a possible role for macrophage-derived cortisol in M1/M2 polarization in common carp. We studied the expression of molecules involved in cortisol synthesis/conversion from and to cortisone like 11β-hydroxysteroid dehydrogenase type 2 and 3. (11β-HSD2 and 3) and 11β-hydroxylase (CYP11b), as well as the expression of glucocorticoid receptors (GRs) and proliferator-activated receptor gamma (PPARγ) in M1 and M2 macrophages. Lastly, we analyzed how inhibition of these molecules affect macrophage polarization. In M1 cells, upregulation of gene expression of GRs and 11β-HSD3 was found, while, in M2 macrophages, expression of 11β-hsd2 was upregulated. Moreover, blocking of cortisol synthesis/conversion and GRs or PPARγ induced changes in expression of anti-inflammatory interleukin 10 (IL-10). Consequently, our data show that carp monocytes/macrophages can convert cortisol. The results strongly suggest that cortisol, via intracrine interaction with GRs, is important for IL-10-dependent control of the activity of macrophages and for the regulation of M1/M2 polarization to finally determine the outcome of an infection.

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Section: Discussionmentioning

confidence: 99%

Section: Inhibition Of Cortisol Conversion and Blocking Of Gr And Pparγmentioning

confidence: 99%

Cortisol Metabolism in Carp Macrophages: A Role for Macrophage-Derived Cortisol in M1/M2 Polarization

Maciuszek

Kłak

Rydz

et al. 2020

IJMS

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“…In Atlantic salmon, researchers found that the diversity of PPAR-γ alleles influenced susceptibility to disease (Sundvold et al, 2010). In orange-spotted grouper, studies indicated that PPAR-γ transcript expression showed a significant increase following Vibrio challenge (Luo et al, 2015). However, the potential role of teleost PPARs in the immune defense system is still not clear.…”

Section: Discussionmentioning

confidence: 99%

Transcriptomics analysis reveals candidate genes and pathways for susceptibility or resistance to Singapore grouper iridovirus in orange-spotted grouper (Epinephelus coioides)

Yang

Wang

et al. 2019

Developmental & Comparative Immunology

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In this study, the transcriptional response of grouper to Singapore grouper iridovirus (SGIV) stimulation was characterized using RNA sequencing. Transcriptome sequencing of three test groups in the grouper was performed using the Illumina MiSeq platform. The three test groups were a control group, which was injected with PBS buffer; a high-susceptible (HS) group, which died shortly after the SGIV injection; and a high-resistance (HR) group, which survived the SGIV injection. In total, 38,253 unigenes were generated. When the HS group was compared with the control group, 885 unigenes were upregulated and 487 unigenes were downregulated. When the HR and control groups were compared, 1114 unigenes were upregulated and 420 were downregulated, and when the HR and HS groups were compared, 1010 unigenes were upregulated and 375 were downregulated. In the KEGG analysis, two immune-related pathways, the p53 and peroxisome proliferator-activated receptor pathways, were detected with highly significant enrichment. In addition, 7465 microsatellites and 22,1569 candidate single nucleotide polymorphisms were identified from our transcriptome data. The results suggested several pathways that are associated with traits of disease susceptibility or disease resistance, and provided extensive information about novel gene sequences, gene expression profiles, and genetic markers. This may contribute to vaccine research and a breeding program against SGIV infection in grouper.

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“…Our pervious studies indicate that the toxicological effects of invading bacteria can induce the oxidative stress that cause DNA damage and lipid peroxidation, while the expression of antioxidant genes (CAT, SOD1, SOD2) serving as biomarkers of oxidative stress increase significantly after vibrio challenge (Luo et al 2015). NADPH oxidase and nitric oxide synthase (NOS) are the two important enzymes related to the free radical generation and the inflammatory response (Jaiswal et al 2000;SATO et al 2002).…”

Section: Discussionmentioning

confidence: 99%

Characterization and expression analysis of Calmodulin (CaM) in orange-spotted grouper (Epinephelus coioides) in response to Vibrio alginolyticus challenge

et al. 2015

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Vibrio alginolyticus containing the highly toxic extracellular product is one of the most serious threats to grouper survival and its minimum lethal dose is approximately 500 CFU/g fish body weight in grouper. To study the toxic effects of V. alginolyticus on the immune system in teleost, Calmodulin (CaM), an important molecular indicator gene, was cloned from the orange-spotted grouper (Epinephelus coioides). The full-length Ec-CaM consisted of a 5'-UTR of 103 bp, an ORF of 450 bp and a 3'-UTR of 104 bp. The Ec-CaM gene encoded a protein of 149 amino acids with an estimated molecular mass of 16.4 kDa and a predicted isoelectric point of 3.93. The deduced amino acid sequence showed that Ec-CaM contained four highly conserved EF-hand domains known to be critical for the function of CaM. Ec-CaM was widely expressed and the highest expression level was observed in liver. Following V. alginolyticus challenge, a sharp increase level of respiratory burst activity and apoptosis ratio were observed. Further analyses of CaM expression and p53 expression in liver, kidney and spleen by qRT-PCR demonstrated that the up-regulated expression of CaM and p53 were observed in the vibrio challenge group. Western blotting analysis confirmed that the Ec-CaM protein was strongly induced in liver at 12 h post-injection, while a sharp increase of p53 protein expression was observed at 24 h post-injection. These results showed CaM expression serving as a potential molecular indicator may help to assess the toxicological effects of V. alginolyticus on the ROS generation and apoptotic process in grouper.

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Molecular cloning, characterization and expression analysis of PPAR gamma in the orange-spotted grouper (Epinephelus coioides) after the Vibrio alginolyticus challenge

Cited by 41 publications

References 66 publications

Cortisol Metabolism in Carp Macrophages: A Role for Macrophage-Derived Cortisol in M1/M2 Polarization

Cortisol Metabolism in Carp Macrophages: A Role for Macrophage-Derived Cortisol in M1/M2 Polarization

Transcriptomics analysis reveals candidate genes and pathways for susceptibility or resistance to Singapore grouper iridovirus in orange-spotted grouper (Epinephelus coioides)

Characterization and expression analysis of Calmodulin (CaM) in orange-spotted grouper (Epinephelus coioides) in response to Vibrio alginolyticus challenge

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