title: Neuritic growth impairment and cell death by unconjugated bilirubin is mediated by NO and glutamate, modulated by microglia, and prevented by glycoursodeoxycholic acid and interleukin-10.
abstract: Neuronal oxidative damage and cell death by unconjugated bilirubin (UCB) showed to be mediated by overstimulation of glutamate receptors and nitric oxide (NO) production, which was abrogated by the bile acid glycoursodeoxycholic acid (GUDCA). Microglia, a crucial mediator of CNS inflammation, evidenced to react to UCB by releasing glutamate and NO before becoming senescent. Our studies demonstrated that neurite outgrowth deficits are produced in neurons exposed to UCB and that conditioned media from these UCB-treated neurons further stimulate NO production by microglia. Nevertheless, microglia protective and/or harmful effects in neonatal jaundice are poorly understood, or unrecognized. Here, we investigated the role of microglia, glutamate and NO in the impairment of neurite sprouting by UCB. Therapeutic potential of the anti-inflammatory cytokine interleukin (IL)-10 and GUDCA was also evaluated. By using MK-801 (a NMDA glutamate-subtype receptor antagonist) and L-NAME (a non-specific NO synthase inhibitor) we found that glutamate and NO are determinants in the early and enduring deficits in neurite extension and ramification induced by UCB. Both GUDCA and IL-10 prevented these effects and decreased the production of glutamate and NO. Only GUDCA was able to counteract neuronal death and synaptic changes. Data from organotypic-cultured hippocampal slices, depleted or non-depleted in microglia, supported that microglia participate in glutamate homeostasis and contribute to NO production and cell demise, which were again abrogated by GUDCA. Collectively our data suggest that microglia is a key player in UCB-induced neurotoxicity and that GUDCA might be a valuable preventive therapy in neonates at risk of UCB encephalopathy.
authors.affiliation: Research Institute for Medicines and Pharmaceutical Sciences (iMed.UL), Faculty of Pharmacy, University of Lisbon, Av. Professor Gama Pinto, 1649-003 Lisbon, Portugal. sandrasilva@ff.ul.pt
authors.forename: Sandra L
authors.initials: SL
authors.lastname: Silva
authors.forename: Ana R
authors.initials: AR
authors.lastname: Vaz
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Diógenes
authors.forename: Nico
authors.initials: N
authors.lastname: van Rooijen
authors.forename: Ana M
authors.initials: AM
authors.lastname: Sebastião
authors.forename: Adelaide
authors.initials: A
authors.lastname: Fernandes
authors.forename: Rui F M
authors.initials: RF
authors.lastname: Silva
authors.forename: Dora
authors.initials: D
authors.lastname: Brites
mesh_terms: D000818:Animals; D001663:Bilirubin / Q000037:antagonists & inhibitors / Q000633:toxicity*; D002417:Cattle; D016923:Cell Death / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D005260:Female; D018698:Glutamic Acid / Q000502:physiology*; D006131:Growth Inhibitors; D006801:Humans; D016753:Interleukin-10 / Q000502:physiology*; D017628:Microglia / Q000187:drug effects / Q000502:physiology; D016501:Neurites / Q000187:drug effects / Q000502:physiology*; D009569:Nitric Oxide / Q000502:physiology*; D009924:Organ Culture Techniques; D011247:Pregnancy; D051381:Rats; D017208:Rats, Wistar; D014580:Ursodeoxycholic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology
chemical_list: D006131:Growth Inhibitors; D016753:Interleukin-10; D009569:Nitric Oxide; D018698:Glutamic Acid; C024033:glycoursodeoxycholic acid; D014580:Ursodeoxycholic Acid; D001663:Bilirubin