No154 Central Hospital Of Chinese Pla

Xinyang, China

No154 Central Hospital Of Chinese Pla

Xinyang, China
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Yan X.-B.,No154 Central Hospital Of Chinese Pla | Hu Z.-H.,No154 Central Hospital Of Chinese Pla | Du Y.-H.,No154 Central Hospital Of Chinese Pla | Lu A.-L.,PLA Fourth Military Medical University | Xing Y.-J.,PLA Fourth Military Medical University
Chinese Journal of Tissue Engineering Research | Year: 2013

Background: Latest researches have shown that P53 can significantly increase the differentiation rate of stem cells by locking the protein pathway P53-P21. While 5-azacytidine can inhibit cell proliferation and lead to apoptosis by activating the P53-P21 protein pathway. Objective: To investigate the effects of P53 specific inhibitor p-fifty three inhibitor-α on differentiation of rat bone marrow mesenchymal stem cells into cardiomyocyte-like cells. Methods: Bone marrow mesenchymal stem cells were separated from Sprague-Dawley rats for culturing and passage. Passage 4 bone marrow mesenchymal stem cells were divided into four groups: normal control group, p-fifty three inhibitor-α group, 5-azacytidine and p-fifty three inhibitor-α+5-azacytidine group. Results and Conclusion: Primary bone marrow mesenchymal stem cells were smaller, and after passaged and induced, the cells were larger than primary cells and exhibited spindle-shaped appearance and arranged in order. MTT assay showed that p-fifty three inhibitor-α at 0-20 μmol/L could reduce the apoptosis of bone marrow mesenchymal stem cells. Identification results of cardiomyocyte-like cells showed that after induced for 4 weeks, little expression of cardiac troponin I and CX-43 could be seen in the normal control group, and the expression of cardiac troponin I and CX-43 in the other three groups was strong. After induced for 4 weeks, the differentiation rate of cardiomyocyte-like cells in the p-fifty three inhibitor-α and p-fifty three inhibitor-α+ 5-azacytidine group was significantly higher than that in the 5-azacytidine group. Western blotting analysis showed that at 1 week after induction, the expression of P53 and P21 in 5-azacytidine group was strongest compared with the other groups, but no expression was observed in p-fifty three inhibitor-α group. At 4 weeks after induction, the expression levels of P53 and P21 in 5-azacytidine group, p-fifty three inhibitor-α group and p-fifty three inhibitor-α+5-azacytidine group were higher than those in the normal control group. The expression levels of P53 and P21 in p-fifty three inhibitor-α group and p-fifty three inhibitor-α+5-azacytidine group at 4 weeks after induction were higher than those at 1 week after induction. It indicates that P53 specific inhibitor p-fifty three inhibitor-α can significantly induce the apoptosis, promote the proliferation of bone marrow mesenchymal stem cells through blocking P53-P21 protein pathway, and inhibit bone marrow mesenchymal stem cells to differentiate into cardiomyocyte-like cells.

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