摘要
目的本研究旨在探讨CCAAT/增强子结合蛋白同源蛋白(CHOP)在小鼠缺血再灌注损伤时的作用,并分析其可能机制。方法 50只C57BL/6J小鼠随机分为5组,每组10只:假手术组(Sham组)、缺血再灌注组(I/R组)、PBS+LipofectamineTM2000转染试剂组(I/R+PBS+Lipo组)、阴性对照组(I/R+siRNASCR组)、I/R+siRNACHOP组。各组检测总肺水含量(TLW)、肺湿干重比(W/D)和肺泡细胞损伤定量指标(IQA),分别用半定量RT-PCR和Western blot检测CHOPmRNA及CHOP蛋白的表达。结果与Sham组相比,I/R组、I/R+PBS+Lipo组和I/R+siRNASCR组W/D、TLW及IQA值升高(P<0.05);经CHOP-siRNA干预后,I/R+siRNACHOP组W/D、TLW及IQA值下降(P<0.01);与Sham组相比,I/R组、I/R+PBS+Lipo组和I/R+siRNASCR组CHOPmRNA及CHOP蛋白水平均表达增加(P<0.01),经CHOP-siRNA干预后,I/R+siRNACHOP组CHOPmRNA及CHOP蛋白水平均下降(P<0.01)。结论 I/R引起小鼠肺组织细胞发生过度的未折叠蛋白反应,并通过CHOP信号转导通路最终导致细胞凋亡,损伤肺组织;抑制CHOP凋亡途径可减轻肺缺血再灌注损伤。
Objective To evaluate the role of CCAAT/enhancer-binding protein homologous protein（ CHOP） in ischemia-reperfusion induced lung injury,and clarify the potential molecular mechanism. Methods Fifty mice of C57 BL /6J were randomly divided into five groups,10 mice in each group,including Sham operation group（ sham group）,ischemia / reperfusion group（ I / R group）,I / R ＋ PBS ＋ Lipofectamine group（ I / R ＋ PBS ＋ Lipo group）,I / R ＋ scramble siRNA group（ I / R ＋ siRNASCR group）,I / R ＋ CHOPsiRNA group（ I / R ＋ siRNACHOP group）. The content of total lung water（ TLW）,wet-to-dry weight ratio（ W/D） of lung and index of quantitative assessment of histologic lung injury（ IQA） were detected,CHOP mRNA and protein expression were detected by RT-PCR and Western blot. Results Compared with Sham group,W / D,TLW and IQA were significantly elevated in I / R group,I / R ＋ PBS ＋ Lipo group and I / R ＋ siRNASCR group（ P〈0. 05）. Moreover,the W / D,TLW and IQA reduced with CHOP-siRNA treatment,respectively（ P〈0. 01）. Compared with Sham group,CHOP mRNA and protein expressions were significantly elevated in I / R group,I / R ＋ PBS ＋ Lipo group and I / R ＋ siRNASCR group,Moreover,the CHOP mRNA and protein expressions reduced with CHOP-siRNA treatment,respectively（ P〈0. 01）.Conclusion I / R could cause excessive unfolded protein response in lung tissue,and induce apoptosis by CHOP signal pathway,damage lung tissue.The inhibition of CHOP pathway could alleviate lung ischemia-reperfusion injury.
出处
《中国生化药物杂志》
CAS
2016年第4期45-48,共4页
Chinese Journal of Biochemical Pharmaceutics
基金
温州市科技局课题(Y20140060)
