ARP-SP基因表达谱芯片对脑损伤的初步研究

目的研究脑挫伤后ARP-SRP基因表达谱差异及其法医学意义。方法从脑组织提取mRNA,应用ARP-SRP-1.0S基因芯片,研究脑挫伤组织与对照脑组织细胞凋亡蛋白和应激反应蛋白相关基因特异性表达差异。结果基因芯片杂交结果中,发现人类溶酶体的胃蛋白酶抑制剂不敏感蛋白酶基因(CLN2)表达水平显著下降。结论CLN2基因与一种婴儿致死性神经变性疾病LINCL有关,但CLN2基因在脑损伤中的作用尚有待进一步研究;基因芯片在研究脑损伤后相关基因的改变具有快速、高通量、高敏度等特点。

A primary study on the ARP-SRP gene expression profiling of brain injury by cDNA microarray

Objective To screen the differential expression of apoptosis-related protein and stress response protein(APR-SRP)genes after human brain injury by cDNA microrarray.Methods The total RNAs were isolated from normal and injured brain tissues of a car-accident victim,and were purified to obtain mRNAs by Oligotex.Both mRNAs from the tissues of the injured and the normal tissue were reversely transcribed to cDNAs with the incorporation of fluorescent dUTP to prepare the hybridization probes.The probes from normal tissue was labeled with Cy3-dUTP,that from the injured tissue with Cy5-dUTP.The mixed probes were hybridized to the BioDoor Chip ARP-SRP-1.0S,a cDNA microarray which contains77apoptosis related protein genes and23stress protein related genes.After high-stringent washing,the cDNA microarray was scanned for the fluorescent signals and showed differences between two tissues.Results Among the100target genes,Only CLN2gene(Homo sapiens lysosomal pepstatin insensitive protease gene)showed distinct deference in expression level between the brain injury and normal tissues.Conclusion cDNA microarray analysis indicated that CLN2gene,which is correlated to a fatal childhood neurodegenerative disease,might be related to the brain injury.The expression level of CLN2gene was significantly decreased in brain injured tissue in comparison to normal tissue.Further analysis of this gene will be helpful to understand the molecular mechanism of brain injury and utilization in forensic medicine.