ghrelin对体外培养中脑神经干细胞细胞活力的影响  

EFFECT OF GHRELIN ON CELL VITALITY OF MIDBRAIN NEURAL STEM CELLS CULTURE IN VITRO

作　　者：宫冰[1] 焦倩[1] 杜希恂[1] 姜宏[1] 

机构地区：[1]青岛大学国家生理学重点(培育)学科,山东青岛266071

出　　处：《青岛大学医学院学报》2016年第2期130-133,共4页Acta Academiae Medicinae Qingdao Universitatis

基　　金：国家自然科学基金项目(31471114);中国博士后科学基金(2015M570571);山东省博士后创新项目专项资金(201401006)

摘　　要：目的探讨不同浓度ghrelin对体外培养的大鼠中脑神经干细胞（NSCs）细胞活力的影响。方法体外培养中脑NSCs并进行鉴定;采用免疫荧光染色检测中脑NSCs上ghrelin受体生长激素促分泌素受体1a（GHSR1a）的表达;以不同浓度的ghrelin对NSCs进行处理,采用CCK-8试剂盒检测NSCs细胞活力的改变。结果分离培养的中脑NSCs可快速增殖形成神经球;培养的细胞经诱导分化后,可分化为β-tubulinⅢ阳性、GFAP阳性和Oligodendrocyte阳性的细胞。体外培养的中脑NSCs细胞膜上GHS-R1a表达阳性。10-9~10-6 mol/L的ghrelin均可提高细胞活力（F=7.293,q=3.636~4.984,P〈0.05）,而[D-Lys3]-GHRP-6可拮抗ghrelin的效应。结论中脑NSCs上表达ghrelin的受体GHS-R1a,ghrelin能够提高体外培养的大鼠中脑NSCs的细胞活力,使细胞存活能力增加。Objective To investigate the effect of ghrelin on vitality of midbrain neural stem cells （NSCs） in rat cultured in vitro. Methods Midbrain NSCs were cultured in vitro and identified. Applying immunofluorescence, the expression of GHS Rla in NSCs was detected and managed with different concentration ghrelin, the vitality of NSCs was assessed using cell counting kit （CCK 8）. Results The NSCs cultured in vitro could quickly form neurospheres. The cultured cells, after induction and dif ferentiation, separated into β tubuli-Ⅲ positive, GFAP positive and Oligodendrocyte positive cells. The cell membrane of NSCs cultured in vitro showed positive GHS-Rla expression, 10^-9, 10^-8, 10^-7 and 10^--6 mol/L gb.relin could improve the vitality of the cells （F=7.293,q=3.636-4.984,P〈0.05）. The effects of ghrelin on NSCs could be antagonized by [D-Lys3]-GHRP-6. Conelusion The ghrelin acceptor GHS-Rla protein is expressed in midbrain neural stem cells. Ghrelin can increase the cell vitality of neural stem cells of rat cultured in vitro.

关 键 词：GHRELIN 中脑 干细胞 细胞培养技术 细胞活力 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]