Salubrinal保护心肌细胞内质网应激凋亡的实验研究

Salubrinal protects endoplasmic reticulum of cardiac muscle cells against stress-associated apoptosis

目的探讨Salubrinal对衣霉素诱导的乳鼠原代心肌细胞凋亡的保护作用及机制。方法采用消化贴壁法获得乳鼠原代心肌细胞,分别给与不同浓度的Salubrinal(10μmol、20μmol、40μmol)预处理,30min后加入TM(5μg/ml),继续培养24h后通过流式细胞和TUNEL检测方法对乳鼠心肌细胞凋亡进行检测;Western印迹法检测凋亡过程中Caspase-12蛋白酶的表达变化。结果经Salubrinal预处理的细胞同单独使用衣霉素相比,细胞凋亡率和细胞凋亡指数均显著下降;Caspase-12表达明显减少。结论Salubrinal对衣霉素诱导的乳鼠原代心肌细胞凋亡具有保护作用,其机制可能是通过抑制内质网应激凋亡通路。

Objective To study the protective effect of Salubrinal on tunicamyiocin（TM）-induced apoptosis of primary cardiac muscle cells in suckling mice and its mechanism.Methods Primary cardiac muscle cells were isolated from mice using the modified 2-step collagenase digestion method.After pretreated with Salubrinal at different concentrations（10μmol,20μmol and 40μmol） for 30min followed by addition of 5μg/ml tunicamyocin（TM）,cardiac muscle cells were incubated for 24h.Apoptosis of the cells was detected by flow cytommetry and TUNEL,respectively.Changes in expression of Caspase-12 during apoptosis were detected by Western blot.Results The apoptosis rate and apoptosis index of cardiac muscle cells and the expression of Caspase-12 in cardiac muscle cells were significantly lower after pretreatment with Salubrinal than after treatment with TM alone.Conclusion Salubrinal protects cardiac muscle cells against TM-induced apoptosis by inhibiting the stress-associated apoptosis pathway in the endoplasmic reticulum.