Effect of neurotrophic factor -3 on the proliferation and apoptosis of bone marrow mesenchymal stem cells and mechanisms

Abstract

Objective To investigate the effect of neurotrophin-3 (NT-3) on the proliferation and apoptosis of bone marrow mesenchymal stem cells (BMSCs) in rats and possible mechanisms. Methods The NT-3 overexpression and lentiviral transfection of BMSCs were co-cultured with neuronal cells respectively and then they were divided into overexpression control group, NT-3 transfection group and shRNA-NT-3 transfection group (NT-3 silencing group). MTT assay was used to detect the cell culture for 24 h, 48 h and 72 h. Cell cycle and apoptosis were detected by flow cytometry for 48 h. Real-time quantitative PCR was used to detect the expression of C/EBPβmRNA.The expression of C/EBPβprotein was detected by Western blot. Results MTT results showed that the proliferation ability of BMSCs in the NT-3 overexpression group was significantly higher than that in the control group (0.650±0.042, 0.826±0.074) at 48 h and 72 h (P<0.05). Compared with the control group (P<0.05), the cell cycle and apoptosis of BMSCs in NT-3 silencing group were significantly decreased at 48 h and 72 h (P<0.05). The results of 48 h cell cycle and apoptosis showed that the percentage of G1 phase in BMSCs was decreased, G2 and S were increased and the apoptosis was decreased. The percentage of G1 phase in G2-S phase and the increase of apoptosis were increased in NT-3 silencing group. The results of Western Blot showed that C/EBPβ mRNA and protein levels were significantly up-regulated in BMSCs of NT-3 overexpression group and significantly decreased in NT-3 silencing group (P<0.05). Conclusion NT-3 may promote the expression of C/EBP beta and affect the expression of its downstream target genes, which can inhibit the apoptosis of BMSCs cells. Key words: Bone marrow mesenchymal stem cells; Neurotrophin-3; CCAAT/enhancer binding proteins β; Neuronal; Apoptosis