lipofectamine and Brain-Ischemia

An efficient gene carrier to the brain is required for successful gene therapy of ischemic stroke. In this study, deoxycholic acid-conjugated polyethylenimine (DA-PEI) was synthesized and evaluated as a heme oxygenase-1 (HO-1) gene carrier for ischemic stroke gene therapy. Gel retardation assay and heparin competition assay showed that DA-PEI formed a stable complex with plasmid DNA. In vitro transfection assays with the luciferase gene showed that DA-PEI had higher transfection efficiency than polyethylenimine (25 kDa, PEI25k) and lipofectamine in Neuro2A cells. Furthermore, DA-PEI had less toxicity than lipofectamine. To evaluate the therapeutic effects of the pβ-HO-1/DA-PEI complex, the complex was injected locally in the brain of the transient middle cerebral artery occlusion animal model. In in vivo studies, DA-PEI was more effective than PEI25k in delivering pβ-HO-1 to the ischemic brain and achieved higher HO-1 expression. As a result, the pβ-HO-1/DA-PEI complexes more effectively reduced infarct volume and the number of apoptotic cells compared with the pβ-HO-1/PEI25k complex. The results suggest that DA-PEI will be useful for HO-1 gene therapy of ischemic stroke.

Topics: Animals; Apoptosis; Brain; Brain Ischemia; Cell Line, Tumor; Deoxycholic Acid; Disease Models, Animal; Gene Transfer Techniques; Genetic Therapy; Heme Oxygenase-1; Humans; Lipids; Male; Mice; Plasmids; Polyethyleneimine; Rats; Rats, Sprague-Dawley; Stroke; Transfection

Vascular endothelial growth factor (VEGF) is most promising in therapeutic angiogenesis for ischemic vascular disease. This paper aimed to study VEGF gene therapy for the treatment of cerebral ischemia. The glial cell was chosen as the target cell for gene transfer, and the expression of VEGF was studied in vitro. VEGF plasmid/liposome complexes were constructed by mixing VEGF plasmid with liposome, and then cultured C6 glioma cells were transfected with these complexes by lipofectamine method. As control, the same kind of cells were exposed to liposome only. Immunohistochemistry was performed to both groups at 24, 48 and 72 hours after transfection. The transfected cells expressed VEGF significantly higher than the control. The present result demonstrated the feasibility of choosing the glial cell as the target cell for VEGF gene transfer, and found the rationale for the cerebral VEGF gene therapy.

Topics: Animals; Brain Ischemia; Brain Neoplasms; Cation Exchange Resins; DNA, Complementary; Endothelial Growth Factors; Feasibility Studies; Genetic Therapy; Genetic Vectors; Glioma; Immunoenzyme Techniques; Lipids; Liposomes; Lymphokines; Neuroglia; Rats; Recombinant Fusion Proteins; Tumor Cells, Cultured; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors