ABSTRACT
AIMS: The aims of this study were to find out whether kallikrein could induce angiogenesis and affect the cerebral blood flow (rCBF) in the early period after cerebral ischemia/reperfusion (CI/R). METHODS: The adenovirus carried human tissue kallikrein (HTK) gene was administrated into the periinfarction region after CI/R. At 12, 24, and 72 h after treatments, neurological deficits were evaluated; expression of HTK and vascular endothelial growth factor (VEGF) were detected by immunohistochemistry staining; the infarction volume was measured; and rCBF was examined by( 14) C-iodoantipyrine microtracing technique. RESULTS: The expression of VEGF was enhanced significantly in pAdCMV-HTK group than controls over all time points (P < 0.05). Furthermore, the rCBF in pAdCMV-HTK group increased markedly than controls at 24 and 72 h after treatment (P < 0.05), and the improved neurological deficit was accompanied by reduced infarction volume in pAdCMV-HTK group 24 and 72 h posttreatment. CONCLUSION: In the early period after CI/R, kallikrein could induce the angiogenesis and improve rCBF in periinfarction region, and further reduce the infarction volume and improve the neurological deficits.
Subject(s)
Cerebrovascular Circulation/genetics , Gene Transfer Techniques , Infarction, Middle Cerebral Artery/therapy , Neovascularization, Physiologic/genetics , Reperfusion Injury/therapy , Tissue Kallikreins/metabolism , Animals , Anti-Inflammatory Agents, Non-Steroidal , Antipyrine/analogs & derivatives , Carbon Isotopes , Cerebral Infarction/etiology , Cerebral Infarction/pathology , Disease Models, Animal , Gene Expression Regulation/genetics , Humans , Infarction, Middle Cerebral Artery/complications , Male , Neovascularization, Physiologic/physiology , Nervous System Diseases/etiology , Nervous System Diseases/therapy , Rats , Rats, Sprague-Dawley , Reperfusion Injury/complications , Statistics, Nonparametric , Time Factors , Tissue Kallikreins/genetics , Tissue Kallikreins/therapeutic use , Vascular Endothelial Growth Factor A/metabolismSubject(s)
Amyloid beta-Peptides/pharmacology , Antipyrine/analogs & derivatives , Free Radical Scavengers/pharmacology , Membrane Potentials/drug effects , Peptide Fragments/pharmacology , Potassium Channels, Voltage-Gated/metabolism , Analysis of Variance , Animals , Antipyrine/pharmacology , Cadmium Chloride/pharmacology , Dose-Response Relationship, Drug , Drug Interactions , Edaravone , Electric Stimulation , PC12 Cells/drug effects , Patch-Clamp Techniques , RatsABSTRACT
To study the potential benefit of the NURR1 gene in Parkinson's disease (PD), we constructed a recombinant republic-deficit adenovirus containing the NURR1 gene (Ad-NURR1) and expressed it in transplanted neural stem cells (NSC). Ad-NURR1 was constructed, and NURR1 mRNA and protein expression were identified by in situ hybridization and western blot analysis, respectively. The identified NURR1 protein could directly or indirectly induce NSC differentiation into neurons. To identify a potential therapeutic use for the transfected NSCs, cells were transplanted into 6-hydroxydopamine lesioned rats. Histopathological and behavioral alterations were evaluated via immunohistochemistry and the ration test, respectively, in rats transplanted with NSCs with or without the Ad-NURR1 adenovirus. The Ad-NURR1 construct effectively expressed the NURR1 protein, which could directly or indirectly induce NSC differentiation into neurons. Both histopathological and behavioral alterations were seen in rats treated with NSCs with or without the Ad-NURR1 construct, although in the case of the latter, the benefits were more robust. These results suggest a potential therapeutic benefit for Ad-NURR1-expressing cells in the treatment of PD. The Ad-NURR1 modification induced NSC differentiation and therefore represents a potential therapy for PD.
Subject(s)
DNA-Binding Proteins/physiology , Genetic Therapy/methods , Neurons/physiology , Parkinson Disease/therapy , Stem Cells/metabolism , Transcription Factors/physiology , Adenoviridae/physiology , Adrenergic Agents/toxicity , Animals , Behavior, Animal , Cell Differentiation/genetics , Cell Line, Transformed , DNA-Binding Proteins/genetics , DNA-Binding Proteins/ultrastructure , Disease Models, Animal , Humans , Microscopy, Electron, Scanning/methods , Nuclear Receptor Subfamily 4, Group A, Member 2 , Oxidopamine/toxicity , Parkinson Disease/etiology , Parkinson Disease/pathology , Rats , Rats, Sprague-Dawley , Stem Cell Transplantation/methods , Time Factors , Transcription Factors/genetics , Transcription Factors/ultrastructure , Transfection , Tyrosine 3-Monooxygenase/metabolismABSTRACT
OBJECTIVE: To study the effects of vascular endothelial growth factor (VEGF) gene transfer on hypoxia-induced apoptosis of neural stem cells in vitro. METHODS: C17.2 neural stem cells cultured in vitro were infected by recombinant adenovirus containing VEGF gene and cultured under hypoxic condition. VEGF expression in these cells was detected by Western blotting, and the apoptotic index was calculated from results of triphosphate-biotin nick end-labeling (TUNEL) assay. Flow cytometry was employed to examine the changes in the cell apoptotic rate after VEGF gene transfer, and the apoptotic bodies were observed under fluorescence microscope with Hoechst33342 staining. RESULTS: The expression of VEGF was significantly increased in pAdCMV VEGF(165)-infected cells, resulting in inhibition of the apoptosis of C17.2 neural stem cells induced by hypoxia manifested by a significantly lower apoptotic rate of the stem cells transfected by pAdCMV VEGF(165) than that of the untransfected cells (10.38%;+/-0.48%; vs 19.98 %;+/-0.55%;, P<0.01) and of the cells transfected with pAdCMV VEGF(165) along with VEGF anti-sense oligodeoxynucleotide (19.07%;+/-0.64%;, <0.01) after hypoxia. CONCLUSIONS: Recombinant adenovirus can efficiently mediate VEGF gene transfer into C17.2 neural stem cells, resulting in high expression of the exogenous VEGF in vitro, which effectively reduces C17.2 neural stem cell apoptosis induced by hypoxia.
