摘要
目的:利用基因表达谱芯片,研究免疫相关基因在衰老过程中表达水平的改变。方法:分别提取20只20月龄SD大鼠和20只4月龄SD大鼠的脾脏mRNA,并通过逆转录制备成杂交探针;老年大鼠用Cy5d-UTP标记,青年大鼠用Cy3 d-UTP标记。将两种探针等量混合后与包含416个免疫相关基因的cDNA芯片进行杂交。通过扫描仪扫描芯片荧光信号和计算机分析,比较老年组和青年组大鼠脾脏的基因表达水平的差异。同步检测老年组大鼠和青年组大鼠的心、肝、肾和血液中的与自由基相关的生化指标。结果:生化指标检测证实20月龄老年大鼠与4月龄青年大鼠清除自由基的能力有显著的差异,说明其生理机能有显著差异,适用于芯片的研究。而芯片结果显示在所研究与免疫相关的基因中,13个基因在老年大鼠脾脏中表达下调,其中包括参与应激和免疫应答的基因,部分参与细胞增殖、分化的调节基因和参与DNA/RNA修复基因;只有1个基因,即编码良性肿瘤淀粉酶的基因在老年大鼠脾脏中表达特异增高。结论:运用基因表达谱芯片可以帮助我们了解免疫系统在衰老过程中的变化,深入理解衰老的发生和发展机制。
Objective: To screen the changes of immune-associate genes expression which is related with the ageing using cDNA microar-ray.Methods:The mRNA from the spleens of young and aged rats were extracted respectively and reversely transcribed to cDNAs with the incorporation of different fluorescent-labeled dUTP as the hybridization probes. The mixed probes were hybridized to the cDNA microarray that contains the cDNA products of 416 immune-associated genes. After high-stringent washing, the cDNA microarray was scanned for the fluorescent signals and showed the gene expression differences between the young and the aged. Some biochemical assays were used to confirm the physiological differences between the young rats and the aged rats. Results: Among the examined genes, 13 down-regulated genes were identified. These genes correlated with immuned response, cell proliferation, cell differentiation and DNA/RNA repair. Only one gene which encoded α-amylase was much higher in the aged than that in the young. Conclusion: Further analysis of the differenially expressed immune-associated genes based on cDNA microarray will be helpful for understanding the molecular mechanism of the ageing.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2002年第8期532-535,共4页
Chinese Journal of Immunology
