hTERT基因转染对人胚胎大脑皮质神经元生长的影响

目的观察hTERT基因转染后对人胚胎大脑皮质神经元生长活力的影响。方法制备pSU-CMV-hTERT腺病毒,以MOI感染人胚胎大脑皮质神经元。免疫细胞化学荧光染色观察神经元内hTERT表达。MTT法测量接种病毒后24h、72h、1周、2周、3周的神经元活力。结果经鉴定正确的重组质粒pSU-CMV-hTERT与骨架质粒正确重组,经包装扩增后得到含hTERT基因的腺病毒。病毒转染24h时荧光染色可见神经元内有hTERT表达,第3周末表达减弱。hTERT基因转染后的神经元平均活力较对照组高,转染病毒后24h、72h、1周、2周、3周,hTERT基因转染对神经元的保护率分别为27.4%、42.4%、17.3%、7.4%和15.9%。结论hTERT基因转染对人胚胎大脑皮质神经元有一定的保护作用。

Influence on human embryonic cortex neuron survival and apoptosis after transfeeted the hTERT gene

Objective To observe the survival autivity of human embryonic cortex neurons,after transfected hTERT gene.Methods Prepare the pSU-CMV-hTERT adenovirus,and infected human embryonic neurons by MOI.Observe hTERT gene,s expression in neurons by immunofluorescence assay.MTT assay was used to determined neurons viability which were infected pSU-CMV-hTERT adenovirus for 24 hours,72 hours,1 week,2 weeks and 3 weeks respectively.Results Identified pSU-CMV-hTERT recombinant plasmid recombined with framework plasmid correctly.After package and amplification,the adenovirus with hTERT gene was obtained.This gene expressed in neurons after transfected for 24 hours,and the expression became fainter at the end of 3 weeks.The average cell viability of neurons which were transfected hTERT gene was higher than the control group.At the time of 24 hours,72 hours,1 week,2 weeks and 3 weeks after being transfected,the protect rates for neurons were 27.4%,42.4%,17.3%,7.4%,15.9% respectively.Conclusion hTERT gene transfection has a certain of protected on humam embryonic corter reweons.