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。
Objective To investigate the effects ot recombinant adenowrus cardiotroptun-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 tiffs 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.（15）. Conclusion CT-1 can alleviate neuron apoptosis anti in,rove cell survival by decreasing caspase-3 expression.
出处
《中华神经外科疾病研究杂志》
CAS
2006年第1期35-39,共5页
Chinese Journal of Neurosurgical Disease Research
