An Sung Su, Jin Hong Lian, Kim Keung Nyun, Kim Dong Seok, Cho Joon, Liu Meng-Lu, Oh Jin Soo, Yoon Do Heum, Lee Min Hyung, Ha Yoon
Department of Neurosurgery, Spine & Spinal Cord Institute, College of Medicine, Yonsei University, Seoul, 120-752, South Korea.
Childs Nerv Syst. 2010 Mar;26(3):323-31. doi: 10.1007/s00381-009-1040-2.
To avoid unwanted adverse effects of higher doses of single treatment of stem cells and gene therapy and increase the therapeutic efficacies, we hypothesized the combined therapy with stem cells and gene therapy. This study assessed the neuroprotective effects of combined gene therapy and stem cell treatment under ischemic hypoxia conditions using hypoxia-inducible vascular endothelial growth factor (VEGF) and bone marrow-derived mesenchymal stem cells (BMSC).
Experimental groups included the control which was N2A cells transfected with empty vectors, the transfection only group which was N2A cells treated with pEpo-SV-VEGF alone, the BMSC only group which was N2A cells transfected with empty vectors and cocultured with BMSCs, and the combined treatment group which was N2A cells treated with pEpo-SV-VEGF and cocultured with BMSCs. Each group was transfected for 4 h and cultured at 37 degrees C and 5% CO2 for 24 h. Each group was then cultivated under hypoxic conditions (1% O2) for 12 h. Neuroprotective effects were assessed by reverse transcription polymerase chain reaction, annexin V, and cytotoxicity assay.
Neurons exposed to hypoxic conditions exhibited neuronal apoptosis. Compared to single treatments, the combined hypoxia-inducible VEGF and BMSC treatment demonstrated a significant increase in VEGF expression and decreased neuronal apoptosis.
These results suggest that combined pEpo-SV-VEGF and BMSC treatment is effective in protecting neurons against hypoxic ischemic injury.
为避免干细胞单次治疗和基因治疗高剂量带来的不良副作用并提高治疗效果，我们设想了干细胞与基因治疗的联合疗法。本研究使用缺氧诱导血管内皮生长因子（VEGF）和骨髓间充质干细胞（BMSC）评估了在缺血缺氧条件下基因治疗与干细胞治疗联合应用的神经保护作用。
实验组包括转染空载体的N2A细胞作为对照组、仅用pEpo-SV-VEGF处理的N2A细胞作为仅转染组、转染空载体并与BMSC共培养的N2A细胞作为仅BMSC组，以及用pEpo-SV-VEGF处理并与BMSC共培养的N2A细胞作为联合治疗组。每组转染4小时，然后在37℃、5%二氧化碳条件下培养24小时。之后每组在缺氧条件（1%氧气）下培养12小时。通过逆转录聚合酶链反应、膜联蛋白V和细胞毒性测定评估神经保护作用。
暴露于缺氧条件下的神经元出现神经元凋亡。与单一治疗相比，联合缺氧诱导VEGF和BMSC治疗显示VEGF表达显著增加，神经元凋亡减少。
这些结果表明，联合pEpo-SV-VEGF和BMSC治疗对保护神经元免受缺氧缺血性损伤有效。
