摘要
目的:探讨SP600125-c-Jun氨基末端激酶(JNK)特异性抑制剂对大鼠肺缺血/再灌注损伤的保护作用及机制。方法:复制在体大鼠原位单肺缺血/再灌注模型,随机分3组(n=10):假手术对照组(Control组)、缺血再灌注组(I/R组)与缺血再灌注+SP600125干预组(SP600125组)。实验结束时取肺组织测湿/干重比(W/D)、肺泡损伤率(IAR);采用蛋白印迹法检测肺组织磷酸化JNK(p-JNK)、JNK蛋白的表达;免疫组化法检测肺组织Bcl-2、Bax、Caspase-3蛋白的表达;原位末端标记法检测肺组织细胞凋亡指数(AI);电镜观察肺组织超微结构的改变。结果:SP600125组肺组织p-JNK、Bax、caspase-3的蛋白表达显著低于I/R组(均P<0.01),Bcl-2的蛋白表达及Bcl-2/Bax的比值显著高于I/R组(均P<0.01),AI、W/D及IAR显著低于I/R组(均P<0.01),肺组织超微结构损伤不同程度减轻。结论:SP600125可能通过抑制JNK信号通路,上调Bcl-2/Bax的比值减少caspase-3依赖性的肺细胞凋亡,从而减轻肺缺血/再灌注损伤。
Objective： To investigate the protective effects and mechanism of SP600125-specificity inhibitor of c-Jun N-terminal kinase （JNK）on lung ischemia/reperfusion injury in rats. Methods： The unilateral lung ischemia/reperfusion model was replicated in vivo. Rats were randomly divided into three groups（ n = 10）： control group, ischemia/reperfusion group（I/R group） and ischemia/reperfusion ＋ SP600125 group （ SP600125 group）. The lung tissues sampled at the end of each experiment were assayed for wet/dry weight ratio （ W/D ）, the injured alveoli rate（IAR）, the expression of phosphorylation JNK （p-JNK） and JNK protein were detected by Western blot, the expression of Bcl-2, Bax, Caspase3 protein were detected by immunocytochemistry techniques, the pnemnocyte apoptosis index （AI） was detected by termi- nal deoxynucleotidyl transferase mediated dUTP nick end abeling（TUNEL）, the ultrastructure changes were observed under electron micro- scope. Results： Compared to I/R group, the expression of p-JNK, Bcl-2, Bax and caspase-3 protein were markedly decreased（all P 〈 0.01）, the expression of Bcl-2 protein and the ratio of Bcl-2/Bax were markedly increased in SP600125 group（all P 〈 0.01 ）. The value of AI, W/D, IAR showed significantly lower than those in I/R group（all P 〈 0.01）. Meanwhile, light morphological and ultrastructure injury were found in SP600125 group. Conclusion： SP600125 can suppress JNK signal pathway, up-regulate the ratio of Bcl-2/Bax to inhibit Cas- pase-3 dependent apoptosis, so that it protects lung tissue from ischemia/reperfusion injury.
出处
《中国应用生理学杂志》
CAS
CSCD
2012年第3期255-258,I0004,共5页
Chinese Journal of Applied Physiology
基金
温州市科技计划项目(Y20070065)
