Pro-inflammatory and pro-oxidant status of pancreatic islet in vitro is controlled by TLR-4 and HO-1 pathways

• Published in: PloS one Vol. 9; no. 10; p. e107656
• Main Authors: Vivot, Kevin, Langlois, Allan, Bietiger, William, Dal, Stéphanie, Seyfritz, Elodie, Pinget, Michel, Jeandidier, Nathalie, Maillard, Elisa, Gies, Jean-Pierre, Sigrist, Séverine
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 24.10.2014; Public Library of Science (PLoS)
• Subjects: Animals; Apoptosis; Biology and Life Sciences; Cell activation; Chemokines; Cobalt; COX-2 inhibitors; Cyclooxygenase 2 - biosynthesis; Cyclooxygenase-2; Cytokines; Diabetes; Gene expression; Heme; Heme Oxygenase-1 - biosynthesis; Heparan sulfate; Humans; Immune system; Implantation; Inflammation; Inflammation - genetics; Inflammation - pathology; Inhibition; Interleukin 6; Interleukin-6 - biosynthesis; Ischemia; Islet cells; Islets of Langerhans - metabolism; Islets of Langerhans - pathology; Islets of Langerhans Transplantation; Laboratory animals; Life Sciences; Macrophages; Macrophages - metabolism; Macrophages - pathology; Medicine and Health Sciences; Migration; NF-κB protein; Oxidative stress; Oxygen; Oxygenase; Pancreas; Pancreatic islet transplantation; Pathogenesis; Phosphorylation; Proteins; Protoporphyrin; Rats; Reactive oxygen species; Reactive Oxygen Species - metabolism; Receptors; Rodents; Signal transduction; Signal Transduction - genetics; Signaling; Target recognition; Therapeutic applications; TLR4 protein; Toll-like receptors; Toll-Like Receptors - biosynthesis; Transplantation; Transplants & implants; France
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0107656

Since their isolation until implantation, pancreatic islets suffer a major stress leading to the activation of inflammatory reactions. The maintenance of controlled inflammation is essential to preserve survival and function of the graft. Identification and targeting of pathway(s) implicated in post-transplant detrimental inflammatory events, is mandatory to improve islet transplantation success. We sought to characterize the expression of the pro-inflammatory and pro-oxidant mediators during islet culture with a focus on Heme oxygenase (HO-1) and Toll-like receptors-4 signaling pathways. Rat pancreatic islets were isolated and pro-inflammatory and pro-oxidant status were evaluated after 0, 12, 24 and 48 hours of culture through TLR-4, HO-1 and cyclooxygenase-2 (COX-2) expression, CCL-2 and IL-6 secretion, ROS (Reactive Oxygen Species) production (Dihydroethidine staining, DHE) and macrophages migration. To identify the therapeutic target, TLR4 inhibition (CLI-095) and HO-1 activation (cobalt protoporphyrin,CoPP) was performed. Activation of NFκB signaling pathway was also investigated. After isolation and during culture, pancreatic islet exhibited a proinflammatory and prooxidant status (increase levels of TLR-4, COX-2, CCL-2, IL-6, and ROS). Activation of HO-1 or inhibition of TLR-4 decreased inflammatory status and oxidative stress of islets. Moreover, the overexpression of HO-1 induced NFκB phosphorylation while the inhibition of TLR-4 had no effect NFκB activation. Finally, inhibition of pro-inflammatory pathway induced a reduction of macrophages migration. These data demonstrated that the TLR-4 signaling pathway is implicated in early inflammatory events leading to a pro-inflammatory and pro-oxidant status of islets in vitro. Moreover, these results provide the mechanism whereby the benefits of HO-1 target in TLR-4 signaling pathway. HO-1 could be then an interesting target to protect islets before transplantation.