Effect of pilose antler polypeptides on apoptosis and membrane stability of mitochondria in cardiac stem cells

doi:10.3969/j.issn.2095-4344.2017.09.021

Abstract

Abstract:

BACKGROUND: Cardiac stem cells can differentiate into cardiomyocytes in vitro under induction of pilose antler polypeptides, which provides a new therapeutic idea for myocardial injury.

OBJECTIVE: To explore the effect of pilose antler polypeptides on the apoptosis rate and membrane stability of mitochondria in cardiac stem cells.

METHODS: We chose healthy male Wistar rats born 2 days to extract cardiac stem cells. The culture dish was used as the experimental unit, and extracted cells were divided into the following four groups (n=12). Blank control group: The same amount of buffer was added for induction; 5-azacytidine group: induced with 5-azacytidine (3 μmol/L); pilose antler polypeptides group: induced with pilose antler polypeptides (800 mg/L); combined group: induced with pilose antler polypeptides (800 mg/L) and 5-azacytidine (3 μmol/L). After 48 hours induction, apoptosis rate of cardiac stem cells in each group was detected with flow cytometry. The membrane potential of mitochondria in cardiac stem cells was detected with immunofluorescence. The expression level of Nkx 2.5, GATA4, ATF-2, and MEF-2C in cardiac stem cells was detected using western blot assay.

RESULTS AND CONCLUSION: There were small, round and bright cells after 2 weeks culture, and cell colonies of cardiac stem cells formed. The apoptosis rate of cardiac stem cells in the 5-azacytidine group increased significantly (P < 0.05) and its membrane potential decreased significantly compared with the blank control group (P < 0.05). The apoptosis rate of cardiac stem cells in the pilose antler polypeptides group and combined group decreased significantly compared with the 5-azacytidine group (P < 0.05, P < 0.01), and their membrane potential increased significantly (P < 0.05, P < 0.01). The results of western blot showed that the expression level of Nkx2.5 increased significantly in the pilose antler polypeptides group and combined group compared with the 5-azacytidine group (P < 0.05), while the expression levels of GATA4 and ATF-2 in each group were low and there were no significant differences among groups (P > 0.05). The expression level of MEF-2C in each group was at a middle level, and there were no significant differences among groups (P > 0.05). To conclude, our experimental findings indicate that pilose antler polypeptides could decrease the apoptosis rate and improve membrane stability of mitochondria in cardiac stem cells. The mechanism may be related to the increased expression of Nkx 2.5, but whether the mechanism is related to GATA4, ATF-2 and MEF-2C needs to be further studied.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Deer, Antlers, Myocytes, Cardiac, Mitochondria, Tissue Engineering

CLC Number:

R394.2

Huang Xiao-wei, Xu Yan, Liu Yue-xin, Li Zhi-meng, Qu Xiao-bo. Effect of pilose antler polypeptides on apoptosis and membrane stability of mitochondria in cardiac stem cells[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(9): 1426-1431.

Figures/Tables 7

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