Influence of CCN3 on differentiation of bone marrow mesenchymal stem cells into endothelial cells / 心血管康复医学杂志

ABSTRACT

Objective:To explore the influence of nephroblastoma over expressed gene (CCN3)on differentiation of bone marrow mesenchymal stem cells (BM-MSCs)into endothelial cells.Methods:BM-MSCs differentiation into endothelial cells induced by condition medium containing vascular endothelial growth factor (VEGF,50ng/ml)was regarded as positive control group.BM-MSCs differentiation into endothelial cells induced by condition medium con-taining recombined CCN3 protein (100 ng/ml)and VEGF (50 ng/ml)was regarded as CCN3 group,and BM-MSCs incubated in pure complete medium was regarded as negative control group.Immunofluorescence staining and semi-quantitative reverse transcription-polymerase chain reaction method (RT-PCR)were used to measure expression of von Willebrand factor (vWF)after 16d to evaluate endothelial cell differentiation,and semi-quantitative RT-PCR method was used to measured expression of Notch1 gene mRNA before and after induced BM-MSCs differentiation. Results:On 16d after induced BM-MSCs differentiation,some cells of positive vWF fluorescence staining were ob-served in positive control group,those of CCN3 group was significantly more than those of positive control group, and expression of vWF mRNA [ratio of optical density (OD) (0.550±0.090)]of CCN3group was significantly higher than that of positive control group (0.358±0.080),(P0.05 all),on 16d after induced differentiation, expressions of Notch1 gene mRNA in positive control group (0.232±0.047)and CCN3 group (0.352±0.029)were significantly higher than that of negative control group (0.132±0.033),P<0.01 all,it of CCN3 group was signifi-cantly higher that of positive control group (P<0.01).Conclusion:CCN3 can enhance BM-BMCs differentiation into endothelial cells by activating Notch1 signal pathway.