Kupffer cell cytokines interleukin-1β and interleukin-10 combine to inhibit phosphoenolpyruvate carboxykinase and gluconeogenesis in cultured hepatocytes
Background and aims: Recent evidence suggests that inflammatory cytokines may mediate reduced hepatic glucose production and reduced blood glucose concentrations in sepsis. Therefore the aim of this study is to provide direct evidence of a cytokine-mediated interaction between Kupffer cells and hepa...
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| Published in | The international journal of biochemistry & cell biology Vol. 36; no. 8; pp. 1462 - 1472 |
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| Main Authors | , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Elsevier Ltd
01.08.2004
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| Subjects | |
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| Abstract | Background and aims: Recent evidence suggests that inflammatory cytokines may mediate reduced hepatic glucose production and reduced blood glucose concentrations in sepsis. Therefore the aim of this study is to provide direct evidence of a cytokine-mediated interaction between Kupffer cells and hepatocytes by characterising the effects of lipopolysaccharide-stimulated Kupffer cells on hepatocyte gluconeogenesis, and the activity of key regulatory enzymes of this pathway.
Methods and results: Primary isolates of hepatocytes co-cultured with lipopolysaccharide-stimulated Kupffer cells in Transwell inserts showed a 48% inhibition of gluconeogenesis (
P<0.001). RNase protection assay and ELISA of Kupffer cells and the culture media following exposure to lipopolysaccharide showed increased levels of interleukin-1 alpha and beta, tumour necrosis factor alpha and IL-10. The addition of IL-1β and IL-10 to hepatocyte cultures inhibited gluconeogenesis by 52% (
P<0.001), whereas each cytokine alone was ineffective. To determine whether altered production or activity of phosphoenolpyruvate carboxykinase or pyruvate kinase was responsible for the reduced glucose synthesis, their mRNA, protein levels and enzyme activities were measured. Primary hepatocytes co-cultured with lipopolysaccharide-stimulated Kupffer cells or cultured with a combination of IL-1β and IL-10 displayed reduced levels of phosphoenolpyruvate carboxykinase mRNA, protein and enzyme activity. In contrast the mRNA, protein levels and enzyme activity of pyruvate kinase were not altered; suggesting that gluconeogenesis was suppressed by downregulation of phosphoenolpyruvate carboxykinase.
Conclusions: Therefore, hypoglycaemia, which is often observed in sepsis, may be mediated by Kupffer cell-derived IL-1β and IL-10. In addition this study suggests these cytokines inhibit phosphoenolpyruvate carboxykinase production and thereby hepatic gluconeogenesis. |
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| AbstractList | Background and aims: Recent evidence suggests that inflammatory cytokines may mediate reduced hepatic glucose production and reduced blood glucose concentrations in sepsis. Therefore the aim of this study is to provide direct evidence of a cytokine-mediated interaction between Kupffer cells and hepatocytes by characterising the effects of lipopolysaccharide-stimulated Kupffer cells on hepatocyte gluconeogenesis, and the activity of key regulatory enzymes of this pathway.
Methods and results: Primary isolates of hepatocytes co-cultured with lipopolysaccharide-stimulated Kupffer cells in Transwell inserts showed a 48% inhibition of gluconeogenesis (
P<0.001). RNase protection assay and ELISA of Kupffer cells and the culture media following exposure to lipopolysaccharide showed increased levels of interleukin-1 alpha and beta, tumour necrosis factor alpha and IL-10. The addition of IL-1β and IL-10 to hepatocyte cultures inhibited gluconeogenesis by 52% (
P<0.001), whereas each cytokine alone was ineffective. To determine whether altered production or activity of phosphoenolpyruvate carboxykinase or pyruvate kinase was responsible for the reduced glucose synthesis, their mRNA, protein levels and enzyme activities were measured. Primary hepatocytes co-cultured with lipopolysaccharide-stimulated Kupffer cells or cultured with a combination of IL-1β and IL-10 displayed reduced levels of phosphoenolpyruvate carboxykinase mRNA, protein and enzyme activity. In contrast the mRNA, protein levels and enzyme activity of pyruvate kinase were not altered; suggesting that gluconeogenesis was suppressed by downregulation of phosphoenolpyruvate carboxykinase.
Conclusions: Therefore, hypoglycaemia, which is often observed in sepsis, may be mediated by Kupffer cell-derived IL-1β and IL-10. In addition this study suggests these cytokines inhibit phosphoenolpyruvate carboxykinase production and thereby hepatic gluconeogenesis. |
| Author | Yeoh, George C.T Olynyk, John K Rigby, Paul J Fournier, Paul A Yerkovich, Stephanie T |
| Author_xml | – sequence: 1 givenname: Stephanie T surname: Yerkovich fullname: Yerkovich, Stephanie T organization: Biochemistry and Molecular Biology, University of Western Australia, Nedlands, 35 Stirling Highway, Crawley 6009, WA, Australia – sequence: 2 givenname: Paul J surname: Rigby fullname: Rigby, Paul J organization: Department of Pharmacology, University of Western Australia, Nedlands, Crawley 6009, WA, Australia – sequence: 3 givenname: Paul A surname: Fournier fullname: Fournier, Paul A organization: School of Human Movement and Exercise Science, University of Western Australia, Nedlands, Crawley 6009, WA, Australia – sequence: 4 givenname: John K surname: Olynyk fullname: Olynyk, John K organization: School of Medicine & Pharmacology, University of Western Australia, Nedlands, Crawley 6009, WA, Australia – sequence: 5 givenname: George C.T surname: Yeoh fullname: Yeoh, George C.T email: yeoh@cyllene.uwa.edu.au organization: Biochemistry and Molecular Biology, University of Western Australia, Nedlands, 35 Stirling Highway, Crawley 6009, WA, Australia |
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| CitedBy_id | crossref_primary_10_1111_bph_13786 crossref_primary_10_1096_fj_202001056RRR crossref_primary_10_1111_iep_12002 crossref_primary_10_1111_j_1742_7843_2006_pto_496_x crossref_primary_10_1152_ajpendo_00345_2017 crossref_primary_10_3390_cells9102258 |
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| Keywords | IFNγ RPA TNFα IL FBPase GK PEPCK Cytokines EGF Phosphoenolpyruvate carboxykinase LPK Kupffer cells ITS LPS Bt 2cAMP G-6-Pase PFK Gluconeogenesis |
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| SubjectTerms | Cytokines Gluconeogenesis Kupffer cells Phosphoenolpyruvate carboxykinase |
| Title | Kupffer cell cytokines interleukin-1β and interleukin-10 combine to inhibit phosphoenolpyruvate carboxykinase and gluconeogenesis in cultured hepatocytes |
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