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| 篇名 |
心肌营养素-1对超氧化损伤神经元的保护作用 |
| 作者 |
杨云华 廖维宏 李红运 |
| 中英文摘要 |
摘要 目的 在体外培养神经元凋亡损伤模型，观察重组腺病毒心肌营养素1（Adv-CT1）对损伤神经元存活的保护作用，了解CT-1对神经元的作用和机制，为神经损伤提供新的治疗措施。 方法 诱导大鼠神经干细胞（NSCs）分化为神经元，建立超氧化诱导神经元凋亡损伤模型，以Adv-CT1转染神经元，应用免疫组化、流式细胞仪凋亡检测等技术，观察CT-1对神经元生长存活的作用以及CT-1和caspase-3基因在损伤神经元表达的变化。 结果 分离培养NSCs，应用无血清小剂量碱性成纤维细胞生长因子（bFGF）神经元培养基诱导NSCs定向分化培养神经元；在神经元凋亡模型中转染Adv-CT1，免疫组化显示神经元中CT-1表达增高（P<0.05，或P<0.01），caspase-3表达降低（P<0.05）；流式细胞检测显示CT-1可减少损伤神经元凋亡比例（P<0.01，或P<0.05），促进细胞存活。 结论 Adv-CT1转染到凋亡神经元后，CT-1表达增加，caspase-3表达降低，提示Adv-CT1对损伤神经元有保护作用，是CT-1通过减少神经元凋亡基因caspase-3表达，抑制凋亡发生，从而促进神经元存活。
Abstract Objective To investigate the effects of recombinant adenovirus cardiotrophin-1 (Adv-CT1) on the survival of neurons in vitro, to observe the protective effects of Adv-CT1 on injured neurons, and to provide solution to the neuronal injury. Methods Neural stem cells (NSCs) were isolated from brains of 14 d rats' embryos and were cultured with conditional medium. NSCs differentiated to neurons in low basic fibroblast growth factor (bFGF) and serum free medium, indicating that this a better method to establish the neuron culture model. The Adv-CT1 was transfected into the H2O2 induced neurons apoptosis injury model. The expression of CT-1 and caspase-3 of injured neurons were checked by immunocytochemistry techniques, and the effects of CT-1 on neurons survival were checked by flowcytometry. Results The Adv-CT1 was transfected into injured neurons, which resulted in the increased expression of CT-1 (P<0.05, or P<0.01). And the expression of caspase-3 was decreased by immunocytochemistry (P<0.05), and ratio of apoptotic neurons decreased by flowcytometry (P<0.01, or P<0.05). Conclusion CT-1 can alleviate neuron apoptosis and improve cell survival by decreasing caspase-3 expression.
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