海马去神经支配损伤后放射状胶质细胞表达RUNX1T1的变化及对向神经元分化的影响

Expression of RUNX1T1 and neuronal differentiation in radial glial cells after deafferented injury of hippocampus in vitro

目的观察海马去神经支配损伤后,海马放射状胶质细胞表达RUNX相关转录因子1转位1(RUNX1T1)和向神经元分化的情况。方法取6只大鼠,采用物理切割大鼠穹隆海马伞术制备海马去神经支配损伤模型,并制备海马提取液,海马放射状胶质细胞体外培养,培养液中加入海马提取液。实验分为去神经支配组、正常组和空白对照组,共6块24孔板,每组各48孔。采用Real-time PCR、Western blotting及免疫荧光技术检测各组放射状胶质细胞表达RUNX1T1 mRNA和蛋白的变化,以及分化成微管相关蛋白2(MAP-2)阳性神经元比例。结果体外培养的海马放射状胶质细胞具有细而长的突起,且表达RUNX1T1,去神经支配组中RUNX1T1阳性细胞的荧光强度高于正常组和空白对照组约1. 8倍,细胞突起也较后两组长。去神经支配组RUNX1T1 mRNA和蛋白表的表达量各上调2. 9倍和2. 4倍,且39. 33%细胞表达MAP-2,与正常组和空白对照组相比,阳性细胞数比例明显增高。结论海马去神经支配损伤后海马放射状胶质细胞上调表达RUNX1T1,并可更多地向神经元分化。

Objective To investigate the expression of runt-related transcription factor 1 translocated to 1( RUNX1 T1) and neuronal differentiation in hippocampal radial glial cells after deafferented injury of hippocampus in vitro.Methods After physical transecting of fimbria-fornix in 6 rats to establish a model of deafferented hippocampus,the extract of hippocampus was prepared. Hippocampal radial glial cells were cultured into 6 pieces of 24 holes cell culture plates in vitro and divided into deafferented injury,normal and blank groups,48 holes in each group. The expression of RUNX1 T1 and proportion of microtubule associated protein 2( MAP-2) positive neurons in each group were detected by immunofluorescence,Real-time PCR and Western blotting. Results Normally hippocampal radial glial cells had thin and long processes and expressed RUNX1 T1. The fluorescence intensity of RUNX1 T1 in deafferented injury group was about1. 8 times higher than that in normal and blank group,and the cell process was also longer. The mRNA and protein expression of RUNX1 T1 in the deafferented injury group were increased by 2. 9 and 2. 4 times respectively,and 39. 33%cells expressed MAP-2. Compared with normal and blank group,the proportion of MAP-2 positive neurons increased more.Conclusion The expression of RUNX1T1 is up-regulated in hippocampal radial glial cells after deafferented injury,and they could differentiate into more neurons in vitro.