异常胆液质载体UC病证大鼠结肠组织炎症相关因子的变化及其机制

发布时间：2019-02-26 13:09

【摘要】：目的:本研究在维吾尔医学(维医)体液论的指导下成功建立异常胆液质载体溃疡性结肠炎(Ulcerative colitis,UC)病证大鼠模型,检测异常胆液质载体UC病证模型组和正常组大鼠结肠组织中IL-1α、IL-1β、iNOS、eNOS等四种炎症相关因子的hnRNA和mRNA表达水平的变化及NF-κB对上述四种炎症相关因子基因转录活性的调控机制,从而阐述NF-κB在异常胆液质载体UC病证发生、发展中的作用机制。方法:根据维医体液论建立异常胆液质载体证候模型的基础上,采用TNBS/乙醇法构建异常胆液质载体UC病证大鼠模型,将动物分为正常组和异常胆液质载体UC病证模型组,应用实时荧光定量逆转录聚合酶链反应(qRT-PCR)方法检测两组大鼠结肠组织中IL-1α、IL-1β、iNOS、e NOS的hnRNA与mRNA表达水平,并分析其表达差异,应用染色质免疫共沉淀(Chromatin Immunoprecipitation,ChIP)-qPCR方法,检测NF-κB与候选基因IL-1α、IL-1β、iNOS、eNOS调控序列的亲和力,阐述存在表达差异的分子机制。结果:1)异常胆液质载体UC病证模型组大鼠体征、症状、结肠粘膜损伤等均符合异常胆液质载体UC病证模型的判定标准;2)qRT-PCR结果显示,与正常组比较,异常胆液质载体UC病证模型组大鼠结肠组织中IL-1α、IL-1β、iNOS的hnRNA表达水平均上调,差异有统计学意义(P0.05),而eNOS的hnRNA表达水平无统计学意义(P0.05);与正常组比较,异常胆液质载体UC病证模型组大鼠结肠组织中IL-1α、IL-1β、iNOS、eNOS的mRNA表达水平均上调,差异有统计学意义(P0.05);3)ChIP-qPCR结果显示,与正常组比较,异常胆液质载体UC病证模型组大鼠结肠组织中NF-κB增强IL-1α、IL-1β、iNOS基因的转录活性,而对eNOS基因转录活性的调控作用不明显。结论:1)异常胆液质载体UC病证模型组大鼠结肠组织中出现免疫功能紊乱;2)异常胆液质载体UC病证模型组大鼠结肠组织中NF-κB通过与IL-1α、IL-1β、iNOS等候选基因调控序列的结合,增强候选基因的转录活性,从而促进候选基因的表达;3)异常胆液质载体UC病证大鼠结肠组织中炎症相关因子表达水平的调控除了NF-κB的转录水平调控作用外,还可能存在RNA的稳定性相关的转录后调控机制。
[Abstract]:Objective: to establish the rat model of ulcerative colitis (Ulcerative colitis,UC) with abnormal bile fluid carrier under the guidance of Uighur medicine (Uighur medicine) theory of body fluid. Detection of IL-1 伪, IL-1 尾, iNOS, in colonic tissue of rats with abnormal choledochal carrier UC disease model group and normal group The changes of hnRNA and mRNA expression levels of eNOS and other four inflammatory related factors and the regulation mechanism of NF- 魏 B on the transcriptional activity of the above four inflammatory related factors were discussed in order to elucidate the occurrence of NF- 魏 B in abnormal choledochal vector UC disease, and the mechanism of the regulation of NF- 魏 B on the transcription activity of these four inflammatory related factors. Mechanisms of action in development Methods: on the basis of establishing the syndrome model of abnormal choledochal carrier according to the theory of Uygur liquid, the rat model of abnormal choledochal carrier UC syndrome was established by TNBS/ ethanol method. The rats were divided into normal group and abnormal choledochal carrier UC syndrome model group, and the rats were divided into two groups: normal group and abnormal choledochal carrier model group. Real-time fluorescence quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect the expression levels of IL-1 伪, IL-1 尾, iNOS,e NOS hnRNA and mRNA in colon tissues of two groups of rats, and the difference was analyzed. Chromatin immunoprecipitation (Chromatin Immunoprecipitation,ChIP)-qPCR was used to detect the affinity of NF- kappa B to the regulatory sequences of candidate genes IL-1 伪, IL-1 尾 and iNOS,eNOS, and to elucidate the molecular mechanism of the differential expression. Results: 1) the signs, symptoms and colonic mucosal injury of rats in the model group of abnormal choledochal carrier UC's disease met the criteria of abnormal choledochal carrier UC's disease model; 2) the results of qRT-PCR showed that the expression of IL-1 伪, IL-1 尾 and iNOS hnRNA in colon tissue of rats in abnormal choledochal carrier UC syndrome group were up-regulated compared with the normal group (P0.05), and there was a significant difference between the two groups (P0.05). The expression level of hnRNA in eNOS was not statistically significant (P0.05). Compared with the normal group, the expression levels of IL-1 伪, IL-1 尾 and iNOS,eNOS mRNA in colon tissue of the model group with abnormal choledochal carrier UC disease were up-regulated (P0.05). 3) the results of ChIP-qPCR showed that NF- 魏 B enhanced the transcriptional activity of IL-1 伪, IL-1 尾 and iNOS genes in colon tissue of rats in abnormal bile carrier UC syndrome group, but did not regulate the transcriptional activity of eNOS gene. Conclusion: 1) in the model group of abnormal choledochal carrier UC's disease, there is an immune disorder in the colon tissue of rats. 2) in the model group of abnormal choledochal vector UC syndrome, NF- kappa B combined with IL-1 伪, IL-1 尾, iNOS and other candidate gene regulatory sequences to enhance the transcriptional activity of candidate genes and thus promote the expression of candidate genes. 3) the regulation of inflammation-associated factor expression in colon of rats with abnormal choledochal carrier UC disease may also have the mechanism of RNA stability-related post-transcriptional regulation, in addition to the regulation of NF- 魏 B transcription level.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R29

【相似文献】