Regional difference in inflammatory response to LPS-injection in the brain: Role of microglia cell density
Pintado Losa, Cristina; Revilla Torres, María Elisa; Vizuete Chacón, María Luisa; García Cuervo, Luisa; Vitorica Ferrández, Francisco Javier; Ruano Caballero, Diego; Castaño Navarro, Angélica
- Published
- 2011
- Publisher
- Elsevier
- Language
- en
Abstract
To elucidate whether density of cells could contribute to the extent of microglial activation, we performed in vitro assays using three different densities of N13 microglia stimulated with LPS. Our results showed that induction of pro-inflammatory factors as TNF-α and iNOS was directly related to cell density, meanwhile the induction of the anti-inflammatory IL-10 was inversely related to cell density. Accordingly, in vivo assays showed that after LPS-injection, iNOS expression was more intense in substantia nigra, a brain area showing specific susceptibility to neurodegeneration after microglia activation, whereas IL-10 expression was more sustained in striatum, an area resistant to damage. These results support that microglia density is pivotal to control the balance between pro- and anti-inflammatory factors release.
Full text
Post‐printof:RegionaldifferenceininflammatoryresponsetoLPS‐injectioninthebrain: Roleofmicrogliacelldensity.JournalofNeuroimmunology238(1‐2):44‐51(2011). FIGURES RegionaldifferenceininflammatoryresponsetoLPS‐injectioninthe brain:roleofmicrogliacelldensity. CristinaPintado1,2,ElisaRevilla1,MaríaL.Vizuete1,2,3,SebastiánJiménez1,2,3,LuisaGarcía‐ Cuervo1,2,3,JavierVitorica1,2,3,DiegoRuano1,2,3andAngélicaCastaño§1,2,3. 1DepartamentodeBioquímicayBiologíaMolecular.FacultaddeFarmacia.Universidadde Sevilla,Spain. 2CentrodeInvestigaciónBiomédicaenRedsobreEnfermedadesNeurodegenerativas (CIBERNED). 3InstitutodeBiomedicinadeSevilla(IBIS)‐HospitalUniversitarioVirgendel Rocío/CSIC/UniversidaddeSevilla.Spain §Correspondingauthor: AngélicaCastaño DepartamentodeBioquímicayBiologíaMolecular.FacultaddeFarmacia.US C/ProfesorGarcíaGonzáleznº2.41012‐Sevilla.Spain. Phonenumber:00‐34‐954556220. E‐mail:[email protected]
Figure 1 control 15 min 1h 6 h 9 h % difference respect to control 0 1000 1500 2000 0.25x106 cells/well 0.50x106 cells/well 1.0x106 cells/well Control 1 h 3 h 6 h 9 h % difference respect to control 0 1000 2000 3000 4000 B iNOS (mRNA) Control 30 m 1 h 3 h 6 h 9 h % difference respect to control 0 2000 4000 6000 8000 10000 A TNF-a (mRNA) C IL-10 (mRNA) a * a * a * a * a * a * a * a * b
Figure 2 6 h 9 h 24 h % difference respect to control 0 100 200 300 400 500 iNOS B * * * 6 h 9 h 24 h C 0.25 0.5 1.0 0.25 0.5 1.0 0.25 0.5 1.0 6 h 9 h 24 h IL-10 (pg/nº cells) x105 0 50 100 150 200 250 0.25x106 cells/well 0.5x106 cells/well 1.0x106 cells/well * * IL-10 C 0.25 0.5 1 0.25 0.5 1 0.25 0.5 1 17kDa 6 h 9 h 24 h C 0.25 0.5 1.0 0.25 0.5 1.0 0.25 0.5 1.0 26kDa 6 h 9 h 24 h % difference respect to control 0 1 2 3 A TNF-a * a a
Figure 3 saline 3h 6h 14h 24h 3 days 7 days Fold increase 0 25 50 75 100 125 TNF-a (mRNA) * A saline 3h 6h 14h 24h 3 days 7 days Fold increase 0 50 100 150 200 250 iNOS (mRNA) a * a * a * B * saline 3h 6h 14h 24h 3 days 7 days Fold increase 0 20 40 60 80 100 120 CD SN a* * * a* saline 3h 6h 14h 24h 3 days 7 days Fold increase 0 20 40 60 80 100 120 CD SN a* * * a* D IL-10 (mRNA) 3h 14 h 24 h 72 h saline SN CD C iNOS protein
GAD65 striatum (mRNA) Saline 14 h 24 h 3 days 7 days % of difference respect to saline 0 50 100 150 200 250 300 Figure 4 GAD65 SN (mRNA) Saline 14 h 24 h 3 days 7 days % of difference respect to saline 0 20 40 60 80 100 120 140 * A B * GAD65 SN (mRNA) 7 days LPS LPS+2,5 nmol SB LPS+3 nmol MT GAD65 mRNA (LPS /Saline) 0.0 0.2 0.4 0.6 0.8 1.0 1.2 * C
