摘要
目的探讨过氧化物氧化还原酶6(Prdx6)对神经干细胞的增殖和分化的影响。方法(1)采用悬浮培养法从新生SD大鼠脑分离培养神经干细胞,传至第三代后进行神经干细胞特异性标记蛋白巢蛋白(Nestin)的免疫荧光检测;(2)细胞实验分组:溶媒组、Prdx61 ng/mL、10 ng/mL、100 ng/mL、1μg/mL组;(3)采用台盼蓝细胞计数法计数各组细胞培养第1 d、第2 d、第3 d、第4 d和第5 d的细胞数,并绘制细胞生长曲线;(4)细胞增殖实验观测各组细胞形成神经球的直径,用Cell Counting kit-8(CCK-8)试剂盒检测各组细胞在450 nm处的吸光度值;(5)在细胞分化实验中,通过免疫荧光染色检测神经干细胞分化后神经元特异性烯醇化酶(neuron specific enolase,NSE)、胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)阳性细胞数。结果(1)体外培养得到悬浮生长的神经干细胞,免疫荧光染色Nestin呈阳性;(2)Prdx6100 ng/mL组和1μg/mL组神经干细胞形成神经球的直径、吸光度值,以及分化后的NSE、GFAP阳性细胞数与溶媒组比较,差异有统计学意义(均P<0.05)。结论Prdx6具有促进神经干细胞增殖和分化的作用。
Objective To investigate the effects of peroxidase 6(Prdx6)on the proliferation and differentiation of neural stem cells.Methods(1)Neural stem cells were isolated from the brain of newborn SD rats by suspension culture and passed to the third generation for immunofluorescence detection of specific protein nestin(Nestin).(2)Cell experiment group:solvent,Prdx61 ng/mL,10 ng/mL,100 ng/mL and 1μg/mL groups.(3)The number of cells on the first day,the second day,the third day,the fourth day,and the fifth day of each group of cells was counted by trypan blue cell counting method and a growth curve was drawn.(4)In the proliferation experiment,the diameter of the neurospheres were compared between the cells of the cell and The Cell Counting kit-8(CCK-8)was used to compare the absorbance at 450 nm of each group of cells.(5)In the cell differentiation experiment,the number of neuronal marker protein neuron specific enolase(NSE)and glial cell marker protein glial fibrillary acidic protein(GFAP)positive cells was compared by immunofluorescence staining.Results(1)In vitro cultured neural stem cell spheres were obtained.The specific marker protein Nestin was positive for immunofluorescence staining.(2)When the concentration of Prdx6 was 100 ng/mL and 1μg/mL,the neurosphere diameter,absorbance value and the number of differentiated NSE and GFAP positive cells in the neural stem cells were statistically significant compared with the solvent group(all P<0.05).Conclusion The proper concentration of Prdx6 can promote the proliferation and differentiation of neural stem cells.
作者
肖广庆
刘希光
伦志刚
陈慧珍
李爱民
XIAO Guang-qing;LIU Xi-guang;LUN Zhi-gang(Graduate School of Xuzhou Medical University,Xuzhou 221004,China;不详)
出处
《临床神经外科杂志》
CAS
2020年第1期60-65,共6页
Journal of Clinical Neurosurgery
关键词
Prdx6
神经干细胞
增殖
分化
Prdx6
neural stem cells
proliferation
differentiation
