内毒素诱导急性肺损伤小鼠基因表达的综合分析

Comprehensive eene expression profile of acute lung injury induced by lipopolysacchande in mice

目的利用改良的基因表达综合分析(SAGE)技术监测内毒素诱发小鼠急性肺损伤的基因表达状况,探讨急性肺损伤的分子机制。方法气管内注射内毒素(25mg/kg)复制急性肺损伤模型。正常对照小鼠气管内注射等体积等渗盐水。内毒素注射24h后以氯胺酮深度麻醉后处死小鼠,整块取出肺脏用于SAGE研究。建立正常和急性肺损伤两个SAGE标记物文库。结果正常文库包括24670个SAGE标记物,代表12168个基因。急性肺损伤文库包括26378个SAGE标记物,代表13397个基因。急性肺损伤组有11种基因升高10倍以上,107种基因升高5-10倍,2121种基因升高2-5倍。7种基因表达降低大于10倍,87种基因降低5-10倍,1571种基因降低2-5倍。结论证实了内毒素诱导的急性肺损伤发生机制中各种基因表达的变化,并发现了以往没有报道的基因。对这些基因功能的进一步研究有助于阐明急性肺损伤的发生机制,并为临床诊断提供有参考价值的血清标记物。

Objective To monitor the systemic gene expression profile in a murine model of li-popolysaccharide (LPS)-induced acute lung injury by the recently modified long serial analysis of gene expression (SAGE) so as to discuss the molecular mechanism of acute lung injury. Methods Acute lung injury was induced by intra-tracheal injection of LPS (25 mg/kg). Control mice were given normal saline in same volume. Animals were killed at 24 hours after the administration of LPS and lungs were harvested en bloc for SAGE study. Results A total of 24 670 tags representing 12 168 transcripts in the control mice and 26 378 tags representing 13 397 transcripts in the mice with lung injury were identified respectively. There were 11 transcripts increased more than 10 folds, 107 transcripts 5-10 folds and 2 121 transcripts 2-5 folds in the LPS-treated mice. But seven transcripts decreased to 1/10, 87 transcripts to 1/10-1/5 and 1 571 transcripts to 1/5-1/2. The most overexpressed genes in the lung injury mice mainly included serum amyloid A 3, metallothionein 2, lipocalin 2, cyclin-dependent kinase inhibitor 1A, lactate dehydrogenase 1 , melatonin receptor, SI00 calcium-binding protein A9 and natriuretic pep-tide precursor. Mitogen activated protein kinase 3, serum albumin, complement component 1 inhibitor, and ATP synthase were underexpressed in the lung injury mice. Conclusion The changes of various genes as well as some unreported genes have been confirmed in the LPS-induced acute lung injury. Further studies of these unreported genes are beneficial to better understanding the mechanism of acute lung injury and may provide useful markers for clinical diagnosis.