【摘要】 目的:探讨钙敏感性受体活化与心肌细胞缺血/再灌注损伤后凋亡及炎症反应的关系。方法:缺氧/复氧法处理新生大鼠心肌细胞,建立心肌缺血/再灌注(ischemia/reperfusion,I/R)模型。新生大鼠心肌细胞随机分为对照组、缺血/再灌注组、GdCl3(CaSR激动剂)组、GdCl3+SB203058(p38MAPK特异抑制剂)组。TUNEL染色法检测心肌细胞凋亡;逆转录聚合酶链式反应(RT-PCR)测定钙敏感性受体(calcium-sensing receptor,CaSR)、肿瘤坏死因子-α(TNF-α)及白细胞介素-6(IL-6)的mRNA表达;免疫蛋白印迹法(Western blot)测定Caspase-3、Bcl-2和丝裂原活化蛋白激酶(p38MAPK)的蛋白表达。结果:模拟I/R增强了CaSR、TNF-α、IL-6、p38MAPK的表达及心肌细胞凋亡;GdCl3进一步增强了上述作用,而对Caspase-3和Bcl-2则分别起着正向及负向调节作用;与GdCl3组相比,GdCl3+SB203058组p38MAPK、TNF-α、IL-6的表达明显降低,而心肌细胞凋亡及CaSR的表达无明显改变。结论:CaSR激活不仅促进新生鼠心肌细胞I/R损伤后心肌细胞凋亡,且通过p38MAPK信号转导促进I/R区域的炎症反应。更多还原

【Abstract】 Objective:To examine whether activation of calcium-sensing receptor(CaSR) is associated with apoptosis and inflammatory reaction of cardiomyocytes stimulated by ischemia/reperfusion(I/R).Methods:Primary neonatal rat ventricular cardiomyocytes were incubated in anoxia/reoxygenation method to establish a model of simulated I/R.And neonatal rat ventricular cardiomyocytes were randomly divided into control group,I/R group,GdCl3(CaSR agonist) group and GdCl3+SB203058(a specific inhibitor of p38MAPK) group.Cardiomyocyte apoptosis was detected by TUNEL assay.The expressions of CaSR,tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) were detected by real reverse transcriptase polymerase chain reaction(RT-PCR).The expressions of Caspase-3,Bcl-2 and p38MAPK were detected by Western blot.Results:The simulated I/R enhances cardiomyocyte apoptosis and the expression of CaSR,TNF-α,IL-6 and p38MAPK.GdCl3 can further increase cardiomyocyte apoptosis and expression of above proteins,along with up-regulation of Caspase-3 and down-regulation of Bcl-2.Compared with GdCl3 group,the mRNA expression of TNF-α and IL-6 were markedly decreased with no significant changes of cardiomyocytes apoptosis and CaSR mRNA in GdCl3+SB203058 group.Conclusion:Activation of CaSR is not only associated with increased cardiomyocytes apoptosis after I/R injury via suppressing Bcl-2 and promoting Caspase-3 expression,but also induces inflammatory reaction via p38MAPK signaling pathway at I/R area.更多还原