Nuclear-transcription factor-κB signal transduction pathway participating in neuronal differentiation of rat marrow mesenchymal stem cells induced by fasudil
10.3760/cma.j.issn.1671-8925.2015.09.008
- VernacularTitle:NF-κB信号通路在法舒地尔诱导大鼠骨髓间充质干细胞向神经元分化中的作用
- Author:
Eryi ZHAO
1
;
Yanjie JIA
;
Daimei WANG
;
Guoqiang WEN
;
Lijun JING
;
Yidong DENG
;
Min GUO
Author Information
1. 海南省人民医院神经内科
- Keywords:
Nuclear-transcription factor-κB;
Fasudil;
Marrow mesenchymal stem cell;
Neuron;
Cell differentiation
- From:
Chinese Journal of Neuromedicine
2015;14(9):899-904
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of nuclear-transcription factor-κB signal transduction pathway on differentiation of rat marrow mesenchymal stem cells (MSCs) into neurons.Methods The MSCs were purified at least four generations in vitro.And then,the MSCs were assigned into three groups:non-serum control group (group A),fasudil treatment group (group B) and fasudil and lipopolysaccharide treatment group (group C).Cells in group A were added serum-free DMEM,those in group B were added induced liquid (DMEM 900 μL+fetal bovine serum 100 μL that contains final concentration of 200 μmol/L of fasudil),and those in group C were added induced liquid (DMEM 900 μL+fetal bovine serum 100 μL that contains final concentration of 200 μmol/1 of fasudil and final concentration of 1000 μg/L of lipopolysaccharide).All the MSCs were induced into neurons for 6 h.The morphology of MSCs was observed under inverted fluorescence microscope.The expressions of neuronspecific enolase (NSE),microtubule-associated protein (MAP)-2,cyclin D1 and glial fibrillary acidic protein (GFAP) in neurons were detected by immunocytochemical method.The mRNA expressions ofcyclin D1 and MA P-2 in neurons were detected by real time-PCR.The protein expressions ofcyclin D1 and MAP-2 were detected by Western blotting.Results (1) The MSCs of group A kept flat and the neurites were scarce;the MSCs of group B and C could differentiate into neurons,and the speed of MSCs of group B differentiating into neurons was faster than that in group C;there was typical neural network in group B and the neural network in group C was scarcer than group B.(2) As compared with those in group A,higher expression levels of NSE and MAP-2,and lower cyclin D1 expression level in group B and group C were noted (P＜0.05);the expression percentage of GFAP was lower than 1％ in both three groups.(3) As compared with group A,group B and group C had significantly lower cyclin D1 protein and mRNA expressions and significantly higher MAP-2 protein and mRNA expressions (P＜0.05).Conclusion The NF-κB signal transduction pathway,by regulating the expression of cyclin D1,participates in the neuronal differentiation of rat marrow mesenchymal stem cells induced by fasudil.
