title: Endotoxin downregulates peroxisome proliferator-activated receptor-gamma via the increase in TNF-alpha release.
abstract: The nuclear receptor peroxisome proliferator-activated receptor-gamma (PPAR-gamma) is anti-inflammatory in a cell-based system and in animal models of endotoxemia. We have shown that PPAR-gamma gene expression is downregulated in macrophages after lipopolysaccharide (LPS) stimulation. However, it remains unknown whether hepatic PPAR-gamma is altered in sepsis and, if so, whether LPS directly downregulates PPAR-gamma. To study this, rats were subjected to sepsis by cecal ligation and puncture (CLP). Hepatic tissues were harvested at 5, 10, and 20 h after CLP. PPAR-gamma gene expression and protein levels were determined by RT-PCR and Western blot analysis, respectively. The results showed that PPAR-gamma gene expression decreased at 10 and 20 h and that its proteins levels were reduced at 20 h after CLP. PPAR-gamma levels were also decreased in animals that were administered LPS. To determine the direct effects of LPS on PPAR-gamma downregulation, LPS binding agent polymyxin B (PMB) was administered intramuscularly after CLP. The administration of PMB significantly reduced plasma levels of endotoxin, but it did not prevent the downregulation of PPAR-gamma expression. We found that circulating levels of TNF-alpha still remained significantly elevated in PMB-treated septic animals. We, therefore, hypothesize that the decrease of PPAR-gamma expression is TNF-alpha dependent. To investigate this, Kupffer cells (KCs) were isolated from normal rats and stimulated with LPS or TNF-alpha. TNF-alpha significantly attenuated PPAR-gamma gene expression in KCs. Although LPS decreased PPAR-gamma in KCs, the downregulatory effect of LPS was blocked by the addition of TNF-alpha-neutralizing antibodies. Furthermore, the administration of TNF-alpha-neutralizing antibodies to animals before the onset of sepsis prevented the downregulation of PPAR-gamma in sepsis. We, therefore, conclude that LPS downregulates PPAR-gamma expression during sepsis via an increase in TNF-alpha release.
authors.affiliation: Department of Surgery, North Shore University Hospital and Long Island Jewish Medical Center and The Feinstein Institute for Medical Research, Manhasset, NY 11030, USA.
authors.forename: Mian
authors.initials: M
authors.lastname: Zhou
authors.forename: Rongqian
authors.initials: R
authors.lastname: Wu
authors.forename: Weifeng
authors.initials: W
authors.lastname: Dong
authors.forename: Asha
authors.initials: A
authors.lastname: Jacob
authors.forename: Ping
authors.initials: P
authors.lastname: Wang
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000906:Antibodies / Q000276:immunology / Q000494:pharmacology; D002478:Cells, Cultured; D004195:Disease Models, Animal; D015536:Down-Regulation / Q000187:drug effects*; D004731:Endotoxins / Q000097:blood / Q000494:pharmacology*; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D007728:Kupffer Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D047495:PPAR gamma / Q000235:genetics / Q000378:metabolism*; D011112:Polymyxin B / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D018805:Sepsis / Q000378:metabolism* / Q000473:pathology; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D000906:Antibodies; D004731:Endotoxins; D008070:Lipopolysaccharides; D047495:PPAR gamma; D014409:Tumor Necrosis Factor-alpha; D011112:Polymyxin B