摘要
研究食欲素-A(Orexin-A)在肝缺血/再灌注(Hepatic ischemia/reperfusion,H-I/R)中表达水平的变化,及其与H-I/R介导的肺损伤的联系,探讨Orexin-A在创伤应激反应中的作用。建立大鼠70%H-I/R模型,实验设计假手术组和不同再灌注时间的损伤组,采用酶比色法检测损伤后血清髓过氧化物酶水平,采用H.E染色和免疫组织化学法分别检测损伤后肺的病理学改变及Orexin-A蛋白表达,同时采用RT-PCR法观察损伤后肺Orexin-A mRNA表达的变化。结果表明:损伤后血清髓过氧化物酶水平有逐步下降的趋势;病理学观察提示H-I/R后期肺损伤显著减轻;与损伤后假手术组相比,四个再灌注组肺Orexin-A蛋白表达均显著降低,缺血60min/再灌注60min(I60’R60’)组、I60’R240’组肺Orexin-A mRNA表达显著降低,而I60’R150’组、I60’R360’组肺Orexin-A mRNA表达显著升高。H-I/R引起Orexin-A表达水平明显的变化,这些变化提示Orexin-A在H-I/R介导的肺损伤中可能发挥一定的负性作用。
To explore the changes of Orexin-A expression levels in hepatic ischemia/reperfusion （H-I/R） and their associations with H-I/R-induced lung injury, and find out the roles of Orexin-A in traumatic stress responses. A 70% H-I/R model of rats was established, setting groups of sham-operation and injury ones with different reperfusion times. Enzyme-colorimetry was used to detect serum myeloperoxidase levels after injury, H. E staining and immunohistochemistry were applied to investigate pathological variations and protein expressions of Orexin-A in lung after injury, respectively, while RT-PCR was used to detect the changes of Orexin-A mRNA expressions in lung after injury. The results showed that serum myeloperoxidase levels expressed a trend to decrease gradually after injury. The pathological investigation suggested that lung injury were significantly alleviated at the later phase of H-I/R. Compared with sham-operation group after injury, protein expressions of Orexin-A in lung of the four reperfusion groups all decreased significantly. The Orexin-A mRNA expressions in lung of 60 min ischemia/60 rain reperfusion （160＇ R60＇） and 160＇ R240＇ groups were significantly lower, while Orexin-A mRNA expressions in lung of 160＇ R150＇ and 160＇ R360＇ groups were significantly higher. H-I/R induces distinct changes of Orexin-A expression levels, and these changes indicate that Orexin-A may exert a certain extent of negative role in H-I/R-induced lung injury.
出处
《标记免疫分析与临床》
CAS
2009年第1期29-33,共5页
Labeled Immunoassays and Clinical Medicine
基金
国家自然科学基金面上项目(30670821)
解放军总医院苗圃基金(06MP83)
