神经免疫调节网络中传入信号和信号传导模式的研究

发布时间：2018-01-29 14:17

本文关键词： 神经免疫调节网络传入通路传入信号白细胞介素-1　出处：《中国协和医科大学》2008年硕士论文　论文类型：学位论文

【摘要】： 神经免疫学是二十世纪末期生命科学领域发展最为迅速的学科之一,神经免疫学科的基础理论“神经—内分泌—免疫调节网络”学说已经得到学术界的认同。在“神经—内分泌—免疫调节网络”研究的基础上,相关的实验证明了神经系统与免疫系统间存在功能调节作用。在神经免疫调节网络信号传导的解剖学构筑的研究中,调节网络的功能相关的传入信号和信号传入通路构成是领域研究争论的焦点之一。本论文在课题组前期不同免疫状态大鼠神经免疫调节功能相关脑区的定位研究工作的基础上,应用蛋白质组学研究中的表面加强激光解析电离—飞行时间—质谱技术(SELDI-TOF-MS)和基质辅助激光解析电离—飞行时间—质谱技术(MALDI-TOF-MS)及双向电泳(2-DE)分离技术等方法对不同免疫状态大鼠的外周血清和脑脊液组份进行比对分析,探索参与神经免疫调节的上传信号及传入通路构成模式,为进一步阐明神经免疫调节网络的解剖学构筑提供实验依据。 SELDI分析结果显示,免疫组动物外周血中观察到221个有显著性差异的信号峰,其中173个为表达量高于对照组的正向表达峰。实验各组大鼠脑脊液的分析结果未见有统计学意义的特异性差异峰表达;在实验组血清中检测到的特异性差异峰也未在脑脊液中观察到相对应的同步变化。在76个最高峰值为免疫2天的免疫功能相关性差异表达的免疫大鼠血清的信号峰中,根据差异信号峰的分子量、pI值和来源细胞在已知的神经介质、神经肽、激素和细胞因子等信号物质中进行分析筛选,推测可能成为神经免疫调节相关的传入信号物质。经2-DE分离和MELDI-TOF-MS进一步分析和鉴定免疫大鼠血清中18KD特异组份,分析报告了18种来源的19个相关肽段序列,共247氨基酸,但相关结果不能满足任何已知物质的氨基酸序列的排序。我们的实验结果证明: 1)本论文采用SELDI技术对不同免疫状态组大鼠和正常对照组大鼠的血清进行对比分析的结果显示实验组表达了221个与免疫功能相关的差异信号峰;差异信号表达峰值的分布在免疫2天的有76个,免疫4天有1个,免疫6天有96个;差异峰值统计学分析具有极显著性差异,某些差异峰的特异性甚至表现为“全或无”的关系。这些实验结果支持了神经免疫调节网络传入信号模式研究中两个重要的代表性假说——“血行传导通路”和“神经传导通路”假说在神经免疫调节网络传入信号通路领域研究中达成的共识,即免疫活性细胞释放功能活动相关信号物质到体液(血液)中,进而启动神经免疫调节功能。 2)对不同实验组大鼠的脑脊液组份对比分析的结果显示,脑脊液中并不存在与免疫功能变化相关的神经免疫调节功能的传入信号物质,血液中免疫功能相关的信号物质的变化没有在脑脊液中平行反映。神经免疫调节功能相关信号传入模式不是“血液—脑脊液—接触脑脊液的神经元系统—中枢脑区”的血行传导方式。本论文神经免疫调节网络信号传导模式研究结果不支持“血行传导通路”假说。 3)根据最高峰值在免疫2天的免疫功能相关的差异信号峰的分子量、PI值和来源细胞,在已知的神经介质、神经肽、激素和细胞因子等信号物质中进行比对筛选,推测细胞因子IL-8、IL-15和IL-18等三个组份可能成为免疫系统释放的跨系统传递的神经免疫调节功能相关的传入信号物质。 4)本论文应用蛋白质组学研究技术的标准方法2-DE-MALDI-TOF-MS技术分析免疫大鼠血清中IL-1分子在神经免疫调节网络传入通路中的地位和作用,发现免疫活性细胞释放到血液中的IL-1不可能成为免疫系统释放的跨系统传递的信号物质,参与神经免疫调节网络功能,否定了“血行传导通路”和“神经传导通路”假说在神经免疫调节网络传入信号通路领域研究中关于IL-1的共识。 5)对免疫大鼠血清双向电泳分离的分子量约18KD,PI约5.0位置差异点进行质谱鉴定的结果显示,该差异点的氨基酸序列不符合数据库收集的任何已知物质的氨基酸序列,提示神经免疫调节网络传入通路的上传信号中,可能包括我们还不了解的新物质。
[Abstract]:Neuroimmunology is one of the most rapidly developing disciplines in the life sciences at the end of the twentieth century . The basic theory of neuroimmunology has been recognized by the academic community . In the study of nerve - endocrine - immune regulation network , the related experiments prove that there is a functional regulation between the nervous system and the immune system . The results of SELDI analysis showed that 221 signal peaks with significant difference were observed in the peripheral blood of the vaccinated group , 173 of which were positive expression peaks in the serum of the experimental group . Our experimental results demonstrate that : 1 ) The results of the comparative analysis of serum from different immune status group rats and normal control rats by SELDI technology show that the experimental group expresses 221 difference signal peaks related to immune function . The distribution of differential signal expression peak is 76 in the two days of immunity , there are 96 in 4 days of immunization and 96 in 6 days of immunization . These experimental results support two important representative hypothesis _ " blood line conduction path " and " nerve conduction path " in the study of the afferent signal pathway of the neural immunomodulatory network . These experimental results support the consensus that the immune active cells release the related signal substance into body fluid ( blood ) and then initiate the neuroimmunoregulation function . 2 ) The results of comparative analysis of cerebrospinal fluid in different experimental groups showed that there were no afferent signal substances related to immune function changes in cerebrospinal fluid . The changes of immune function - related signal substances in the blood were not reflected in the cerebrospinal fluid . 3 ) according to the molecular weight , PI value and source cell of the difference signal peak related to the immune function of the two - day immune function according to the highest peak value , three components such as IL - 8 , IL - 15 and IL - 18 in known nerve media , neuropeptides , hormones , cytokines and the like are compared and screened , and three components such as cytokines IL - 8 , IL - 15 and IL - 18 are presumed to be an incoming signal substance related to the neuroimmunoregulation function of the cross - system transmission released by the immune system . 4 ) In this paper , the status and function of IL - 1 molecule in the serum of immune rats were analyzed by using the standard method 2 - DE - MALDI - TOF - MS . It was found that IL - 1 released into the blood could not be a signal substance to be released by the immune system . 5 ) The results show that the amino acid sequence of the difference point does not conform to the amino acid sequence of any known substance collected in the database , suggesting that the amino acid sequence of the difference point does not conform to the amino acid sequence of any known substance collected by the database , suggesting that the uploading signal of the afferent pathway of the neural immunomodulatory network may include new substances that we don ' t know yet .

【学位授予单位】：中国协和医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392

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