摘要
目的筛选与束缚应激大鼠应激反应恢复速度相关的基因,探讨应激反应个体恢复的分子机制。方法观察大鼠2小时束缚应激后血浆促肾上腺皮质激素(adrenocorticotropic-hormone,ACTH)及皮质酮的变化规律,根据应激结束后1小时的血浆ACTH及皮质酮下降程度将大鼠分为快速恢复组与慢速恢复组。采用微阵列技术检测快速恢复组与慢速恢复组下丘脑组织基因表达的差异,用实时逆转录-聚合酶链反应验证部分基因的表达差异。结果基因芯片分析结果显示:大部分基因在两组间并无差异表达,在11个差异表达的基因中,快速恢复组中参与整合素信号通路的踝蛋白(talin)、整合素α6和丝/苏氨酸蛋白磷酸酶PP1β催化亚基(serine/threonine pro-tein phosphatase PP1-beta catalytic subunit,PP1B)基因有1.5倍上调,而结合粘附分子1(junctional adhesion mol-ecule 1, F11r)基因有 1.5 倍下调。实时逆转录 - 聚合酶链反应结果与芯片分析结果一致。结论本研究构建了束缚应激大鼠恢复过程的下丘脑基因表达谱,结果提示整合素信号通路可能参与应激的恢复过程。
Objective To search novel genes or pathways involved in the recovery process after restraint stress in rats. Methods We compared the hypothalamus transcriptional profiles of two different recovery patterns (fast recovery vs slow recovery) from restraint stress in rats using oligonucleotide microarray, the recovery pattern was determined by the decrement of plasma adrenocorticotropic-hormone (ACTH) and corticosterone levels during one hour recovery period after stress. A real-time quantitative RT-PCR was applied to validate the differential expressed genes. Results Analysis of the microarray data showed that most of genes were not differentially expressed between fast recovery group and slow recovery group. Among the differentially expressed genes we found that talin, together with serine/threonine protein phosphatase PPl-beta catalytic subunit (PP-1B) and integrin α-6 precursor (VLA-6) genes, were at least 1.5 fold upregulated in the fast recovery group, while junctional adhesion molecule 1 (F11r) was 1.5 fold down-regulated in the fast recovery group. Conclusion The results implied that integrin signaling pathway may be involved in the recovery from restraint stress in rats. The present study provided a global overview of hypothalamus transcriptional profiles during the process of recovery from the restraint stress in rats. The integrin signaling pathway seems to be involved in the recovery process, which deserves further study to clarify the integrin-mediated recovery mechanism after restraint stress.
关键词
束缚应激
促肾上腺皮质激素
皮质酮
应激后恢复
基因芯片
physical restraint
adrenocorticotropic hormone
corticosterone
post-stress recovery
oligonucleotide microarray