Transient transfection of human CDNF gene reduces the 6-hydroxydopamine-induced neuroinflammation in the rat substantia nigraReportar como inadecuado

Transient transfection of human CDNF gene reduces the 6-hydroxydopamine-induced neuroinflammation in the rat substantia nigra - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

Journal of Neuroinflammation

, 11:209

First Online: 16 December 2014Received: 12 September 2014Accepted: 25 November 2014


BackgroundThe anti-inflammatory effect of the cerebral dopamine neurotrophic factor CDNF was shown recently in primary glial cell cultures, yet such effect remains unknown both in vivo and in 6-hydroxydopamine 6-OHDA models of Parkinson’s disease PD. We addressed this issue by performing an intranigral transfection of the human CDNF hCDNF gene in the critical period of inflammation after a single intrastriatal 6-OHDA injection in the rat.

MethodsAt day 15 after lesion, the plasmids p3xNBRE-hCDNF or p3xNBRE-EGFP, coding for enhanced green florescent protein EGFP, were transfected into the rat substantia nigra SN using neurotensin NTS-polyplex. At day 15 post-transfection, we measured nitrite and lipoperoxide levels in the SN. We used ELISA to quantify the levels of TNF-α, IL-1β, IL-6, endogenous rat CDNF rCDNF and hCDNF. We also used qRT-PCR to measure rCDNF and hCDNF transcripts, and immunofluorescence assays to evaluate iNOS, CDNF and glial cells microglia, astrocytes and Neuron-Glial type 2 NG2 cells. Intact SNs were additional controls.

ResultsIn the SN, 6-OHDA triggered nitrosative stress, increased inflammatory cytokines levels, and activated the multipotent progenitor NG2 cells, which convert into astrocytes to produce rCDNF. In comparison with the hemiparkinsonian rats that were transfected with the EGFP gene or without transfection, 6-OHDA treatment and p3xNBRE-hCDNF transfection increased the conversion of NG2 cells into astrocytes resulting in 4-fold increase in the rCDNF protein levels. The overexpressed CDNF reduced nitrosative stress, glial markers and IL-6 levels in the SN, but not TNF-α and IL-1β levels.

ConclusionOur results show the anti-inflammatory effect of CDNF in a 6-OHDA rat of Parkinson’s disease. Our results also suggest the possible participation of TNF-α, IL-1β and IL-6 in rCDNF production by astrocytes, supporting their anti-inflammatory role.

KeywordsNG2 cells Astrocytes NBRE promoter Neurotrophic factor CDNF Polyplex Cytokines Nitrosative stress Abbreviations4-HEA4-hydroxyalkanal

6-OHDA6- hydroxydopamine

ANOVAanalysis of variance

BDNFbrain-derived neurotrophic factor

bpbase pair

BSAbovine serum albumin

CDNFcerebral dopamine neurotrophic factor

C-EBPβCCAAT-enhancer-binding protein β



EDTAethylenediaminetetraacetic acid

EGFPenhanced green fluorescent protein

EGTAethylene glycoltetraacetic acid

ELISAenzyme-linked immunosorbent assay

ERendoplasmic reticulum

ERK-MAPKmitogen-activated protein kinase

GDNFglial cell line-derived neurotrophic factor

GFAPglial fibrillary acidic protein



iNOSinducible nitric oxide synthase


MANFmesencephalic astrocyte-derived neurotrophic factor


MFImean fluorescence intensity


NBREnerve growth factor responsive element

NFnuclear factor

NG2Neuron-Glial type 2

NGFneural growth factor

NOnitric oxide

NTFneurotrophic factor


OX42marker for brain microglia

PBPparabranchial pigmented

PBSphosphate-buffered saline

PDParkinson’s disease

qRT-PCRquantitative Reverse Transcription- Polymerase Chain Reaction

RAGsrecombinant activator genes


ROIregion of interest

SNsubstantia nigra

SNcsubstantia nigra pars compacta

SNrsubstantia nigra pars reticulata

SNc-SNrinter-region between substantia nigra pars compacta and pars reticulata

THtyrosine hydroxylase

TNF-αtumor necrosis factor alpha

Electronic supplementary materialThe online version of this article doi:10.1186-s12974-014-0209-0 contains supplementary material, which is available to authorized users.

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