正常及饥饿大鼠胃壁组织基因表达谱的研究

目的通过荧光芯片技术从整体水平研究正常及饥饿大鼠胃壁组织基因表达谱,探讨饥饿大鼠胃壁组织基因表达的变化。方法用SD大鼠建立正常和饥饿两种模型,动物断头取胃壁组织抽提RNA,荧光芯片杂交,并对实验结果采用real-timePCR、Northern blot和Western blotting方法验证。结果在由基因或表达序列标签(ESTs)组成的9233个杂交点的大鼠基因芯片上,共有504个在饥饿大鼠胃壁组织呈差异表达。其中263个为已知基因,87个为已知的ESTs。在差异表达的基因或ESTs中,上调262个,下调242个,这其中包括与细胞信号转导、细胞结构运动等相关的基因或ESTs。经初步验证,芯片数据的可靠性达80%。结论利用荧光芯片技术研究正常及饥饿大鼠胃壁组织的基因表达谱,可以大规模、高通量发现胃在饥饿状态发生改变的基因,为进一步阐明饥饿胃肠生理、寻找新的与能量代谢及食欲调节相关的基因以及发现基因的新功能奠定基础。

Differential analysis on gene profiles of gastric tissues in normal and fasting rats

Objective To study changes of gene profiles of gastric tissue in normal and fasting rats by using cDNA chip.Methods SD rats were used to establish fasting rat models and sacrificed to take RNA in gastric tissues.Northern blot,Wester blotting and real-time PCR were applied to detect expressions of genes of gastric tissue in fasting or normal rats.Results After screening of 9 233 genes or expressed sequence tags(ESTs),504 genes or ESTs were identified showing differential expression in fasting rat gastric tissue,with 263 known genes and 87 known ESTs.Among the differential expression genes,262 genes were up-regulated and 242 genes were down-regulated,which including the genes associated with cell structure and movement and cell signal transduction.The reliability of the chip was 80% through the validation of real-time PCR. Conclusion Gene chip is a good method to detect the different expression genes in fasting rats gastric tissue,which provides us more information for gastric physiology,new energy metabolism and appetite adjustment genes,and new function of genes.