γ-分泌酶抑制剂对正常乳鼠心肌细胞的影响  

The effects of DAPT treatment on cardiomyocytes from neonatal rats

作　　者：杨阳 段维勋 周京军 毕生辉 杨建宝 冯建宇 侯晓薇 金振晓 易定华 

机构地区：[1]第四军医大学第一附属医院心血管外科,西安710032 [2]第四军医大学第一附属医院生理学教研室,西安710032

出　　处：《中国体外循环杂志》2011年第2期89-92,共4页Chinese Journal of Extracorporeal Circulation

基　　金：陕西省自然科学基金(2007C248);西京医院学科助推计划(XJZT09M16)

摘　　要：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 effects of N-（N-（3,5-difluorophenacetyl）-1-alanyl）-S-phenylglycine t-butyl ester（DAPT） treatment on normal cardiomyocytes from neonatal rats.METHODS Cardiomyocytes from neonatal rats were separated and cultured in vitro.The cells were incubated with different concentrations（in μmol/L： 10,5,2.5,1.25,0.63,0.31,0.16） of DAPT for 24 h,or incubated with DAPT（5 μmol/L） for 1 h,2 h,4 h,8 h,16 h,24 h and 48 h.The viability of the cells was investigated with MTT method.The expression of Notch intracellular domain（NICD） and Bcl-2 protein were detected with Western-Blotting.RESULTS The viability of neonatal rat cardiomyocytes was significantly depressed with DAPT incubation.The effects were positively related to the concentrations of DAPT and the time of incubation.DAPT treatment also significantly decreased the expression of NICD and Bcl-2 protein.CONCLUSION DAPT can interrupt the transduction of Notch signaling pathway in neonatal rat cardiomyocytes and inhibit the viability of the cells and increase their apoptosis.

关 键 词：乳鼠 心肌细胞 DAPT NICD BCL-2 

分 类 号：R541[医药卫生—心血管疾病]