论文部分内容阅读
目的探讨Notch信号特异性阻断剂γ-分泌酶抑制剂(DAPT)对正常乳鼠心肌细胞的影响。方法 SD乳鼠心肌细胞体外分离培养后,与不同浓度DAPT(10μmol/L、5μmol/L、2.5μmol/L、1.25μmol/L、0.625μmol/L、0.3125μmol/L、0.15625μmol/L)孵育24 h,或者与DAPT(5μmol/L)孵育不同时间(1 h、2 h、4 h、8 h、16 h、24 h、48 h),MTT法测定心肌细胞活力;Western Blotting方法测定心肌细胞Notch1受体胞内区(NICD)和Bcl-2蛋白表达量。结果 DAPT显著抑制正常乳鼠心肌细胞的存活,同时浓度越高、时间越长,其抑制效果越明显;DAPT处理后心肌细胞NICD和Bcl-2表达均降低。结论 DAPT可阻断Notch信号通路,抑制正常乳鼠心肌细胞的增殖,促进细胞凋亡。
Objective To investigate the effect of Notch signal inhibitor γ-secretase inhibitor (DAPT) on normal neonatal rat cardiomyocytes. Methods SD rat cardiomyocytes were isolated and cultured in vitro and incubated with different concentrations of DAPT (10μmol / L, 5μmol / L, 2.5μmol / L, 1.25μmol / L, 0.625μmol / L, 0.3125μmol / L, 0.15625μmol / L) 24 h, or incubated with DAPT (5 μmol / L) for different time (1 h, 2 h, 4 h, 8 h, 16 h, 24 h, 48 h). The viability of cardiomyocytes was measured by MTT assay. Notch1 receptor intracellular domain (NICD) and Bcl-2 protein expression. Results DAPT significantly inhibited the survival of normal neonatal rat cardiomyocytes. At the same time, the higher the concentration, the longer the inhibitory effect was. The expressions of NICD and Bcl-2 were decreased after DAPT treatment. Conclusion DAPT blocks Notch signaling pathway, inhibits the proliferation of normal neonatal rat cardiomyocytes and promotes apoptosis.