Analysis of the effect of stress on the gut-associated immune systems by the genetic method

据报道，压力与肠道疾病的发展密切相关，如大肠癌、炎症性肠病和肠易激综合征。因此，我们研究了隔离应激对肠道相关淋巴系统的影响。应激急性期肠系膜淋巴结细胞的ConA反应增强，慢性期则被抑制。T/B比值在急性期升高，在慢性期降低。IL-4的产生在慢性期明显增强，显示Th2型显性的可能性。由此可见，应激对肠系膜淋巴结细胞的影响显著，而对脾脏细胞的影响呈反比关系。血清可的松水平在急性期上调，P物质水平在慢性期上调。利用DNA阵列技术进行基因发现分析，分析应激对大肠的影响。与对照组相比，基因的上调信号在慢性期最为丰富。IL- 12分泌减少，IL- 10及其受体表达上调，提示应激改变了Th1/Th2平衡。在分离2天和30天的样本中，许多与激素、神经递质及其受体相关的基因上调。这些结果表明，分离应激对大肠产生了深刻的影响，并导致了神经元的超敏反应。实验采用三硝基氯苯（TNBS）诱导的慢性溃疡性结肠炎小鼠，分别在隔离应激负荷下和非隔离应激负荷下进行。从大肠中提取RNA，进行基因表达分析。结果表明，TNBS治疗小鼠的转录因子和dna结合蛋白相关基因上调。离子通道和转运蛋白相关基因在TNBS处理和负荷应激小鼠中表达上调。因此，应激增加并调节大肠中的基因表达。这些结果由RT-PCR和real-time PCR确定。少

It is reported that stress is deeply concerned with the development of intestinal diseases, such as large intestine cancer, an inflammational bawel disease, and irritable colon syndrome. Therefore, we investigated the effect of the isolation stress on the gut-associated lymphoid system. ConA responses of mesenteric lymph node cells were reinforced at the acute phase of the stress, and was suppressed at the chronic phase. T/B ratio was enhanced at the acute phase, and suppressed at the chronic phase. The production of IL-4 was markedly enhanced at the chronic phase, showing the possibility of Th2 type-dominant. Thus, the mesenteric lymph node cells were notably influenced by the stress, though the spleen cells exhibited the inverse relationship. Serum cortisone levels were upregulated in acute phase, while substance P levels were upregulated in chronic phase.The influence of stress on the large intestine was analyzed in the gene discovery analysis using DNA array. Gene's upregulated sig … More nificantly compared to control were most abundant in chronic phase. Reduction of IL- 12 production and upregulation of IL- 10 and its receptor suggested that the stress changes the Th1/Th2 balance to Th2. In the sample from 2 and 30 day isolation, many genes related to hormone, neurotransmitter, and their receptors were upregulated. These results indicated that isolation stress profoundly affects the large intestine, and made it to the neuronal hypersensitivity.Next experiments were performed using chronic ulcerative colitis-induced mice by trinitrochlorobenzene (TNBS) under isolation stress load for 30 days or not. RNA was extracted from the large intestines and gene expression analysis was performed. As a result, the transcription factor & DNA-binding proteins related genes were upregulated in TNBS treated mice. The ion channels & transport proteins related genes were upregulated in mice treated with TNBS and loaded with stress. Thus, stress increases and modulates gene expression in large intestine. These results were ascertained by RT-PCR using the real-time PCR. Less