Effect of pilose antler polypeptides on apoptosis and membrane stability of mitochondria in cardiac stem cells
10.3969/j.issn.2095-4344.2017.09.021
- VernacularTitle:鹿茸多肽对心肌干细胞凋亡和线粒体膜稳定性的影响
- Author:
Xiaowei HUANG
;
Yan XU
;
Yuexin LIU
;
Zhimeng LI
;
Xiaobo QU
- From:
Chinese Journal of Tissue Engineering Research
2017;21(9):1426-1431
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Cardiac stem cells can differentiate into cardiomyocytes in vitro under induction of pilose antlerpolypeptides, 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 ofmitochondria in cardiac stem cells.METHODS: We chose healthy male Wistar rats born 2 days to extract cardiac stem cells. The culture dish was used asthe experimental unit, and extracted cells were divided into the following four groups (n=12). Blank control group: Thesame amount of buffer was added for induction; 5-azacytidine group: induced with 5-azacytidine (3 μmol/L); pilose antlerpolypeptides group: induced with pilose antler polypeptides (800 mg/L); combined group: induced with pilose antlerpolypeptides (800 mg/L) and 5-azacytidine (3 μmol/L). After 48 hours induction, apoptosis rate of cardiac stem cells ineach group was detected with flow cytometry. The membrane potential of mitochondria in cardiac stem cells wasdetected with immunofluorescence. The expression level of Nkx 2.5, GATA4, ATF-2, and MEF-2C in cardiac stem cellswas detected using western blot assay.RESULTS AND CONCLUSION: There were small, round and bright cells after 2 weeks culture, and cell colonies ofcardiac 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). Theapoptosis rate of cardiac stem cells in the pilose antler polypeptides group and combined group decreased significantlycompared 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 thepilose antler polypeptides group and combined group compared with the 5-azacytidine group (P < 0.05), while theexpression 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 significantdifferences among groups (P > 0.05). To conclude, our experimental findings indicate that pilose antler polypeptidescould decrease the apoptosis rate and improve membrane stability of mitochondria in cardiac stem cells. Themechanism 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.
