摘要
【目的】探讨重组腺病毒介导血管内皮生长因子(VEGF)165基因对缺氧状态下对神经干细胞凋亡的作用。【方法】体外培养C17.2神经干细胞,建立神经干细胞缺氧模型;将携带人类VEGF基因的重组腺病毒感染C17.2神经干细胞;Western-blot法检测VEGF蛋白的表达;流式细胞术测定细胞凋亡率;Hoechst33342染色荧光显微镜观察凋亡小体。【结果】转染pAdCMVVEGF165的C17.2神经干细胞成功表达VEGF蛋白;缺氧后神经干细胞凋亡率为(19.98%±0.55%),转染pAdCMVVEGF165后的细胞凋亡率为(10.38%±0.48%,P<0.01),而转染pAdCMVVEGF165+VEGF反义寡核脱氧核酸经干细胞凋亡率为(19.07%±0.64%),与对照组无显著差异(P>0.05)。【结论】腺病毒介导的VEGF165基因能成功表达VEGF;外源性VEGF对C17.2神经干细胞具有抗凋亡作用,能够提高C17.2神经干细胞对缺氧的耐受性,有利于神经干细胞的生存。
Objective] To investigate the effect of vascular endothelial growth factor (VEGF) gene on the apoptosis of neural stem cell after hypoxia in vitro. [Methods] C17.2 neural stem cell was cultured and infected by recombinant adenovirus containing VEGF gene, then cultured under hypoxic condition; VEGF expression was detected by Western blot analysis, the apoptotic ratio was inspected by flow cytometry, and the apoptosis bodies was observed by Hoechst33342 staining under fluorescence microscope. [Results] The expression of VEGF in pAdCMV VEGF165 infected cells was significantly increased. The apoptotic ratio of neural stem cells after hypoxia was (19.98% ± 0.55%). The apoptotic ratio of neural stem cells infected by pAdCMV VEGF165 after hypoxia was (10.38% ± 0.48%, P< 0.01), the apoptotic ratio of neural stem cells infected by pAdCMV VEGF165 and anti-sense oligodeoxynucleotide after hypoxia was (19.07%±0.64%, P< 0.01). [Conclusions] Recombinant adenovirus containing VEGF gene can transfer C17.2 neural stem cell efficiently and the exogenous VEGF expressed highly in vitro. Recombinant adenovirus media VEGF gene transfer could reduce C17.2 neural stem cell apoptosis after hypoxia.
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2005年第3期293-296,共4页
Journal of Sun Yat-Sen University:Medical Sciences
基金
广东省卫生厅科研基金资助项目(B2004047)
关键词
神经干细胞
血管内皮生长因子
凋亡
腺病毒载体
缺氧
neural stem cell
vascular endothelial growth factor
apoptosis
adenovirus vector
hypoxia
