用微阵列方法分离+GZ反复暴露后大鼠心脏组织应激性差异表达基因

目的筛选 Gz反复暴露后大鼠心脏组织特异性应激基因。方法提取 Gz反复暴露后1h及对照组大鼠心脏组织总RNA，分离纯化Poly A RNA，与Atlas Rat Stress Array膜杂交，经半定量RT-PCR进行差异基因表达的特异性鉴定。结果初步得到10个差异表达基因，选取HSP70、HSP60、HSPB2、HO-1、HMG2、LCAD作为候选基因，经半定量RT-PCR做特异性验证，发现 Gz反复暴露后1h，HSPB2、HO-1和LCAD mRNA水平升高，HSP70和HMG2 mRNA水平下调，HSP60表达水平未发生改变。结论HSPB2、HO-1是 Gz反复暴露后大鼠心脏组织中早期重要的保护性物质；应用Atlas Rat Stress Array为研究 Gz反复暴露对心脏组织的作用机制提供新的思路和方法。

Isolation of the differentially expressed stress genes of heart tissue from rat after repeated + Gz exposures with microarray analysis

Objective To isolate the specific stress genes from heart of the rat underwent repeated G z exposures. Methods Total RNA was isolated from heart of rats 1 h after repeated 10 G z exposures and that of controls. Poly A RNA was extracted and hybridized with Atlas Rat Stress Array, and semi quantitative RT PCR was used to corroborate the results of Atlas experiment. Results Ten differentially expressed genes were preliminarily isolated from heart of rat underwent repeated G z exposures, and HSP70, HSP60, HSPB2, HO 1, HMG2 and LCAD genes were selected as candidates for relative specific genes. The specificity of these expressions were further confirmed by semi quantitative RT PCR HSPB2, HO 1, LCAD mRNA levels were found to be increased in rat heart after repeated G z exposures, while HSP70 and HMG2 mRNA levels decreased, and HSP60 expression did not changed. Conclusion HSPB2, HO 1 are important early protective agents in heart of rat underwent repeated G z exposures. Atlas Rat Stress Array can provide a novel method to reveal the heart stress induced by repeated G z exposures.