摘要
目的建立体内和体外缺血再灌注(ischemia/reperfusion,I/R)模型,观察缺血再灌注心肌细胞凋亡情况及STAT3蛋白表达变化;检测sRAGE对缺血再灌注心肌细胞凋亡及STAT3的蛋白表达的影响。方法复制C57BL/6J小鼠心脏和原代心肌细胞缺血再灌注模型,在sRAGE和(或)STAT3抑制剂AG490的干预下,通过检测TUNEL及caspase-3活性评价心肌细胞凋亡的程度;通过Western blotting检测磷酸化的STAT3(p-STAT3)及总的STAT3(t-STAT3)蛋白的表达。结果体内实验,与Sham组相比,I/R组TUNEL阳性细胞数目和caspase-3活性分别增加了115%和120%,I/R组p-STAT3/STAT3比值降低了50%,sRAGE降低了I/R诱导的心肌细胞凋亡,包括TUNEL阳性细胞数目降低了51%,caspase-3活性降低了36%,此外,sRAGE预处理I/R组的p-STAT3/STAT3比值增加了381%;体外实验,与Control组相比,I/R组TUNEL阳性细胞数目和caspase-3活性分别增加了380%和77%,I/R组p-STAT3/STAT3比值降低了69%,sRAGE(900 ng/m L)同样降低了I/R诱导的心肌细胞凋亡,表现为TUNEL阳性细胞数目降低了63%,caspase-3活性降低了33%,此外,sRAGE预处理I/R组的p-STAT3/STAT3比值增加了243%,与I/R+sRAGE组相比较,I/R+sRAGE+AG490组的TUNEL阳性细胞数目升高了126%,caspase-3活性增加了42%,p-STAT3/STAT3比值降低了68%。结论 sRAGE可通过激活STAT3抑制缺血再灌注诱导的心肌细胞凋亡。
Objective To test the effect of sRAGE on myocardial apoptosis and STAT3 protein expression with or without STAT3 inhibitor AG490 following ischemia/reperfusion in vivo and in vitro. Methods C57BL/6J mice undergone left anterior descending coronary artery ligation were used as in vivo model and neonatal rat cardiomyocyte subjected to ischemic buffer as an in vitro model. Apoptosis was detected by TUNEL staining and caspase-3 activity. Expression of STAT3/p-STAT3 protein were detected by Western blotting analysis in the presence and absence of the JAK2 inhibitor AG 490. Results In vivo, compared with sham group, the number of TUNEL positive cells and caspase-3 activity were increased by 115% and 120%, and the ratio of p-STAT3/STAT3 was reduced by 50% ; sRAGE （100 txg/day） reduced the TUNEL-positive myocytes by 51%, and activity of caspase-3 by 36%, increased the ratio of p- STAT3/STAT3 by 381% followed by I/R. In vitro, compared with control group, the number of TUNEL positive cells and caspase-3 activity increased by 380% and 77%, and the ratio of p-STAT3/STAT3 was reduced by 69%, sRAGE （900 ng/mL） reduced the TUNEL-positive myocytes by 63%, and caspase-3 activity by 33%, increased the ratio of p-STAT3/STAT3 by 243% followed by I/R. The effect of sRAGE reduction on TUNEL-positive myocytes and caspase-3 activity, raise of the ratio of p-STAT3/STAT3 were attenuated by STAT3 inhibitor AG490. Conclusion These results suggest that sRAGE protects cardiomyocytes from apoptosis induced by I/R in vitro and in vivo by activating STAT3.
出处
《首都医科大学学报》
CAS
北大核心
2016年第1期41-47,共7页
Journal of Capital Medical University
基金
国家自然科学基金(81570321
81370313)
北京市科技新星计划(2010B050)
北京市卫生系统高层次卫生技术人才培养计划(2013-3-046)资助项目~~
