Glucagon Reduces Neutrophil Migration and Increases Susceptibility to Sepsis in Diabetic Mice
Sepsis is one of the most common comorbidities observed in diabetic patients, associated with a deficient innate immune response. Recently, we have shown that glucagon possesses anti-inflammatory properties. In this study, we investigated if hyperglucagonemia triggered by diabetes might reduce the m...
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Frontiers Media S.A.
2021-07-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2021.633540/full |
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| author | Daniella Bianchi Reis Insuela Maximiliano Ruben Ferrero Cassiano Felippe Gonçalves-de-Albuquerque Cassiano Felippe Gonçalves-de-Albuquerque Amanda da Silva Chaves Adriano Yagho Oliveira da Silva Hugo Caire Castro-Faria-Neto Hugo Caire Castro-Faria-Neto Rafael Loureiro Simões Thereza Christina Barja-Fidalgo Patricia Machado Rodrigues e Silva Marco Aurélio Martins Adriana Ribeiro Silva Adriana Ribeiro Silva Vinicius Frias Carvalho Vinicius Frias Carvalho |
| author_facet | Daniella Bianchi Reis Insuela Maximiliano Ruben Ferrero Cassiano Felippe Gonçalves-de-Albuquerque Cassiano Felippe Gonçalves-de-Albuquerque Amanda da Silva Chaves Adriano Yagho Oliveira da Silva Hugo Caire Castro-Faria-Neto Hugo Caire Castro-Faria-Neto Rafael Loureiro Simões Thereza Christina Barja-Fidalgo Patricia Machado Rodrigues e Silva Marco Aurélio Martins Adriana Ribeiro Silva Adriana Ribeiro Silva Vinicius Frias Carvalho Vinicius Frias Carvalho |
| author_sort | Daniella Bianchi Reis Insuela |
| collection | DOAJ |
| description | Sepsis is one of the most common comorbidities observed in diabetic patients, associated with a deficient innate immune response. Recently, we have shown that glucagon possesses anti-inflammatory properties. In this study, we investigated if hyperglucagonemia triggered by diabetes might reduce the migration of neutrophils, increasing sepsis susceptibility. 21 days after diabetes induction by intravenous injection of alloxan, we induced moderate sepsis in Swiss-Webster mice through cecum ligation and puncture (CLP). The glucagon receptor (GcgR) antagonist des-his1-[Glu9]-glucagon amide was injected intraperitoneally 24h and 1h before CLP. We also tested the effect of glucagon on CXCL1/KC-induced neutrophil migration to the peritoneal cavity in mice. Neutrophil chemotaxis in vitro was tested using transwell plates, and the expression of total PKA and phospho-PKA was evaluated by western blot. GcgR antagonist restored neutrophil migration, reduced CFU numbers in the peritoneal cavity and improved survival rate of diabetic mice after CLP procedure, however, the treatment did no alter hyperglycemia, CXCL1/KC plasma levels and blood neutrophilia. In addition, glucagon inhibited CXCL1/KC-induced neutrophil migration to the peritoneal cavity of non-diabetic mice. Glucagon also decreased the chemotaxis of neutrophils triggered by CXCL1/KC, PAF, or fMLP in vitro. The inhibitory action of glucagon occurred in parallel with the reduction of CXCL1/KC-induced actin polymerization in neutrophils in vitro, but not CD11a and CD11b translocation to cell surface. The suppressor effect of glucagon on CXCL1/KC-induced neutrophil chemotaxis in vitro was reversed by pre-treatment with GcgR antagonist and adenylyl cyclase or PKA inhibitors. Glucagon also increased PKA phosphorylation directly in neutrophils in vitro. Furthermore, glucagon impaired zymosan-A-induced ROS production by neutrophils in vitro. Human neutrophil chemotaxis and adherence to endothelial cells in vitro were inhibited by glucagon treatment. According to our results, this inhibition was independent of CD11a and CD11b translocation to neutrophil surface or neutrophil release of CXCL8/IL-8. Altogether, our results suggest that glucagon may be involved in the reduction of neutrophil migration and increased susceptibility to sepsis in diabetic mice. This work collaborates with better understanding of the increased susceptibility and worsening of sepsis in diabetics, which can contribute to the development of new effective therapeutic strategies for diabetic septic patients. |
| first_indexed | 2024-12-22T02:12:16Z |
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| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-22T02:12:16Z |
| publishDate | 2021-07-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-400650b86144462f986fc1a2b58b3c972022-12-21T18:42:22ZengFrontiers Media S.A.Frontiers in Immunology1664-32242021-07-011210.3389/fimmu.2021.633540633540Glucagon Reduces Neutrophil Migration and Increases Susceptibility to Sepsis in Diabetic MiceDaniella Bianchi Reis Insuela0Maximiliano Ruben Ferrero1Cassiano Felippe Gonçalves-de-Albuquerque2Cassiano Felippe Gonçalves-de-Albuquerque3Amanda da Silva Chaves4Adriano Yagho Oliveira da Silva5Hugo Caire Castro-Faria-Neto6Hugo Caire Castro-Faria-Neto7Rafael Loureiro Simões8Thereza Christina Barja-Fidalgo9Patricia Machado Rodrigues e Silva10Marco Aurélio Martins11Adriana Ribeiro Silva12Adriana Ribeiro Silva13Vinicius Frias Carvalho14Vinicius Frias Carvalho15Laboratory of Inflammation, Oswaldo Cruz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, BrazilLaboratory of Inflammation, Oswaldo Cruz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, BrazilLaboratory of Immunopharmacology, Oswaldo Cruz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, BrazilLaboratory of Immunopharmacology, Biomedical Institute, Federal University of the State of Rio de Janeiro, Rio de Janeiro, BrazilLaboratory of Inflammation, Oswaldo Cruz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, BrazilLaboratory of Immunopharmacology, Oswaldo Cruz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, BrazilLaboratory of Immunopharmacology, Oswaldo Cruz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, BrazilLaboratory of Inflammation, National Institute of Science and Technology on Neuroimmunomodulation (INCT-NIM), Rio de Janeiro, BrazilLaboratory of Cellular and Molecular Pharmacology, Biology Institute, State University of Rio de Janeiro, Rio de Janeiro, BrazilLaboratory of Cellular and Molecular Pharmacology, Biology Institute, State University of Rio de Janeiro, Rio de Janeiro, BrazilLaboratory of Inflammation, Oswaldo Cruz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, BrazilLaboratory of Inflammation, Oswaldo Cruz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, BrazilLaboratory of Immunopharmacology, Oswaldo Cruz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, BrazilLaboratory of Inflammation, National Institute of Science and Technology on Neuroimmunomodulation (INCT-NIM), Rio de Janeiro, BrazilLaboratory of Inflammation, Oswaldo Cruz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, BrazilLaboratory of Inflammation, National Institute of Science and Technology on Neuroimmunomodulation (INCT-NIM), Rio de Janeiro, BrazilSepsis is one of the most common comorbidities observed in diabetic patients, associated with a deficient innate immune response. Recently, we have shown that glucagon possesses anti-inflammatory properties. In this study, we investigated if hyperglucagonemia triggered by diabetes might reduce the migration of neutrophils, increasing sepsis susceptibility. 21 days after diabetes induction by intravenous injection of alloxan, we induced moderate sepsis in Swiss-Webster mice through cecum ligation and puncture (CLP). The glucagon receptor (GcgR) antagonist des-his1-[Glu9]-glucagon amide was injected intraperitoneally 24h and 1h before CLP. We also tested the effect of glucagon on CXCL1/KC-induced neutrophil migration to the peritoneal cavity in mice. Neutrophil chemotaxis in vitro was tested using transwell plates, and the expression of total PKA and phospho-PKA was evaluated by western blot. GcgR antagonist restored neutrophil migration, reduced CFU numbers in the peritoneal cavity and improved survival rate of diabetic mice after CLP procedure, however, the treatment did no alter hyperglycemia, CXCL1/KC plasma levels and blood neutrophilia. In addition, glucagon inhibited CXCL1/KC-induced neutrophil migration to the peritoneal cavity of non-diabetic mice. Glucagon also decreased the chemotaxis of neutrophils triggered by CXCL1/KC, PAF, or fMLP in vitro. The inhibitory action of glucagon occurred in parallel with the reduction of CXCL1/KC-induced actin polymerization in neutrophils in vitro, but not CD11a and CD11b translocation to cell surface. The suppressor effect of glucagon on CXCL1/KC-induced neutrophil chemotaxis in vitro was reversed by pre-treatment with GcgR antagonist and adenylyl cyclase or PKA inhibitors. Glucagon also increased PKA phosphorylation directly in neutrophils in vitro. Furthermore, glucagon impaired zymosan-A-induced ROS production by neutrophils in vitro. Human neutrophil chemotaxis and adherence to endothelial cells in vitro were inhibited by glucagon treatment. According to our results, this inhibition was independent of CD11a and CD11b translocation to neutrophil surface or neutrophil release of CXCL8/IL-8. Altogether, our results suggest that glucagon may be involved in the reduction of neutrophil migration and increased susceptibility to sepsis in diabetic mice. This work collaborates with better understanding of the increased susceptibility and worsening of sepsis in diabetics, which can contribute to the development of new effective therapeutic strategies for diabetic septic patients.https://www.frontiersin.org/articles/10.3389/fimmu.2021.633540/fullcAMPdiabetesglucagonneutrophilsepsis |
| spellingShingle | Daniella Bianchi Reis Insuela Maximiliano Ruben Ferrero Cassiano Felippe Gonçalves-de-Albuquerque Cassiano Felippe Gonçalves-de-Albuquerque Amanda da Silva Chaves Adriano Yagho Oliveira da Silva Hugo Caire Castro-Faria-Neto Hugo Caire Castro-Faria-Neto Rafael Loureiro Simões Thereza Christina Barja-Fidalgo Patricia Machado Rodrigues e Silva Marco Aurélio Martins Adriana Ribeiro Silva Adriana Ribeiro Silva Vinicius Frias Carvalho Vinicius Frias Carvalho Glucagon Reduces Neutrophil Migration and Increases Susceptibility to Sepsis in Diabetic Mice Frontiers in Immunology cAMP diabetes glucagon neutrophil sepsis |
| title | Glucagon Reduces Neutrophil Migration and Increases Susceptibility to Sepsis in Diabetic Mice |
| title_full | Glucagon Reduces Neutrophil Migration and Increases Susceptibility to Sepsis in Diabetic Mice |
| title_fullStr | Glucagon Reduces Neutrophil Migration and Increases Susceptibility to Sepsis in Diabetic Mice |
| title_full_unstemmed | Glucagon Reduces Neutrophil Migration and Increases Susceptibility to Sepsis in Diabetic Mice |
| title_short | Glucagon Reduces Neutrophil Migration and Increases Susceptibility to Sepsis in Diabetic Mice |
| title_sort | glucagon reduces neutrophil migration and increases susceptibility to sepsis in diabetic mice |
| topic | cAMP diabetes glucagon neutrophil sepsis |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2021.633540/full |
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