摘要
背景:神经干细胞具有自我更新和多向分化潜能,但正常情况下,神经干细胞数目太少,且大部分处于静息状态,促进其增殖是神经干细胞治疗神经退行性疾病的关键。目的:观察蛇床子素对体外培养的神经干细胞增殖作用的影响,分析其促进增殖能力的作用机制。方法:体外培养神经干细胞,在增殖培养基中培养至第3代,加入不同浓度的蛇床子素(10,50和100μmol/L)作用24 h用CCK-8法检测细胞活力,再培养3,5,7 d后测量神经球半径,用免疫荧光组化法计数Ki67阳性细胞数目;再培养3 d后利用RT-PCR技术检测神经干细胞中Notch 1、Hes 1和Mash 1基因表达的情况。结果与结论:与正常组比较,50,100μmol/L的蛇床子素具有明显促进神经干细胞增殖能力的作用,100μmol/L作用最为显著,可引起Notch 1基因、Hes 1基因上调表达,对Mash 1基因基本无影响,说明蛇床子素对体外培养的神经干细胞具有促进增殖作用,其作用机制可能与激活Notch信号通路上的Notch 1、Hes 1基因有关。
BACKGROUND： Neural stem cells have self-renewal and multidirectional differentiation potential, but under normal circumstances, the number of neural stem cells is less, and most cells are in the resting state. Thus, to promote the proliferation of neural stem cells is the key to the treatment of neurodegenerative diseases. OBJECTIVE： To investigate the effects of osthole on the proliferation of neural stem cells cultured in vitro, and to analyze its mechanism underlying promoting the proliferation. METHODS： Neural stem cells were cultured in vitro, and passage 3 cells were cultured with different concentrations of osthole（10, 50 and 100 μmol/L）. After 24 hours, cell vitality was determined by cell counting kit-8. After 3, 5, 7 days of further culture, the radius of neurospheres was measured, and Ki67-positive cells were counted by immunofluorescence staining. Meanwhile, after 3 days of further culture, the gene expression of Notch 1, Hes 1 and Mash 1 in neural stem cells was detected by RT-PCR. RESULTS AND CONCLUSION： Compared with the control group, 50, 100 μmol/L osthole could obviously promote the proliferation ability of neural stem cells. 100 μmol/L osthole had the most significant effect and increased the expression of Notch 1 gene, Hes 1 gene, but it had no effect on Mash 1 gene. These results suggest that osthole can promote proliferation of neural stem cells cultured in vitro and its mechanism may be associated with activation of Notch 1 gene and Hes 1 gene in Notch signaling pathway.
出处
《中国组织工程研究》
CAS
CSCD
2014年第32期5184-5189,共6页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金(81173580)
国家自然科学基金资助对外交流与合作项目(81210108050)~~
