不同应激源对机体免疫反应影响的差异性及相关机制

发布时间：2018-06-04 19:58

本文选题：应激 + 免疫应答　；参考：《大连医科大学》2005年硕士论文

【摘要】：应激(stress)反应是人类日常生活中不可避免的,创伤、高热、工作压力、应试等均是其常见的应激源。一般将应激定义为:机体对不协调状态或内环境稳定受到威胁时所做出的反应。亚急性应激或慢性应激,一般指超过 24 小时的持续或反复应激,主要伴有神经、内分泌、免疫和行为等变化。处于应激状态的躯体可产生一系列的生理、心理、内分泌、神经生化及免疫功能等方面的改变,这是机体受到应激而做出的适应性全身应答,借以维持机体内环境的相对稳定。同时,随着社会的发展,人们意识到亚急性应激比急性应激更能严重影响人类的身心健康。所以,亚急性应激在疾病的发生、发展中所起的作用也越来越受到关注。但国内有关这方面的研究尚处于刚刚起步阶段,尤其在不同应激源对机体免疫系统是否有影响、有何影响的方面还没有系统详尽的研究报道。已有研究表明,位于小鼠的 17 号染色体上的 H-2 复合体(Histocompatibility 2 complex, H-2 complex),即主要组织相容性复合体(Major Histocompatibility Complex, MHC),是多态性最丰富的基因系统。H-2 复合体 I 区中分立的基因编码小鼠 II 类分子的α和β链。其中最主要的为 A 基因和 E 基因。A 亚区的基因多态性低于 E 亚区。有研究表明,绝大多数种系小鼠均表达 MHC II 类 I-Ab 基因,此为种系生存的必需基因。因此,本实验即选用多态性相对低的 A 亚区为目标区,通过对 A b 亚区内恒定基因片段的检测,分析研究不同应激源对 ·2· 机体免疫反应影响的差异性,并试图找出其相关机制。本文建立了不同于急性应激的小鼠亚急性束缚应激模型(倾向于心理应激模型)与亚急性热应激模型(倾向于躯体应激模型),并于造模第 3、10、16 天对小鼠进行卵清蛋白抗原免疫,于两组小鼠各死亡 1/3 时进行尾静脉采血,至死亡 2/3 时止,每天采血一次,共计 7 天。在可比的条件下用酶联免疫吸附实验(ELISA)对不同性质应激模型组小鼠抗体激发水平进行动态检测,分析亚急性心理应激与亚急性躯体应激对免疫应答的影响;同时建立了小鼠 9 天亚急性应激动物模型, 对不同应激组及对照组小鼠血液中皮质醇进行定量检测;借助异硫氰荧光素(FITC)标记的 H-2IAb 单克隆抗体,运用流式细胞术,对不同应激小鼠淋巴细胞表面 H-2IAb 的表达进行定量检测;同时采用一步法 RT-PCR 的方法检测 H-2IAb 基因表达水平;于第 1、5 天对 9 天亚急性应激动物模型小鼠进行卵清蛋白抗原免疫,通过测定小鼠的体重变化, 分析体重变化率与免疫指标的关系。结果显示两种亚急性应激对体液免疫应答的影响存在显著性差异 (p0.05),7 天的检测中,束缚应激组抗卵清蛋白抗体水平(U)低于热应激组;皮质醇检测结果显示两应激组小鼠血液中皮质醇含量 (nmol/L)均高于正常对照组,但无统计学差异(束缚应激组 5.57±1.75, 热应激组 4.20±1.36,对照组 3.91±0.99,p0.05);流式细胞仪检测结果显示不同应激组小鼠淋巴细胞表面 IAb 的表达(Mean.)有显著性差异,束缚应激组显著低于热应激组(2.86±1.76,6.83±2.32,p0.05); RT-PCR 电泳结果显示所有小鼠均检测出 H-2 IAb 基因,有肉眼可见的强度区别,同时,与流式细胞仪检测结果相关(相关系数 r=0.649), 证明了流式细胞仪中单克隆抗体的准确性;在小鼠免疫指标(U)发生变化的同时(束缚应激组 31.89±28.92,热应激组 76.06±72.71,对照组 87.44±57.35),三组小鼠间体重变化率存在显著性差异(束缚应激组 -0.09±0.06,热应激组-0.01±0.05,对照组 0.11±0.06),同时排除了免疫指标与体重变化率之间的相关性。通过分析得出以下结论:1、本实验成功建立了不同性质亚急性应激动物模型,即倾向于心理应激的亚急性束缚应激模型与倾向于躯体
[Abstract]:Stress (stress) reactions are unavoidable in human daily life. Trauma, high fever, work stress, and examination are all common stressors. Stress is generally defined as the response of the body to incongruity or the stability of the internal environment. Subacute stress or chronic stress generally refers to more than 24 hours of continuous or recurrent stress. Stress, mainly accompanied by changes in nerve, endocrinology, immunity and behavior. The body in a stressful state can produce a series of changes in physiology, psychology, endocrinology, neurobiochemistry and immune function. This is the adaptive systemic response that the body is subjected to stress to maintain the relative stability of the environment in the body. It is realized that subacute stress can seriously affect human physical and mental health more seriously than acute stress. Therefore, subacute stress has attracted more and more attention in the occurrence and development of disease. However, the domestic research on this aspect is still in the beginning stage, especially on the immune system of different stressors. There are no systematic and detailed reports on the influential and influential aspects.
It has been shown that the H-2 complex (Histocompatibility 2 complex, H-2 complex) located on chromosome 17 of mice, namely, the major histocompatibility complex (Major Histocompatibility Complex, MHC), is the alpha and beta chain of the gene encoding mouse II classes in the most polymorphic gene system.H-2 complex I region. The most important one is that the gene polymorphism of the A gene and the.A subregion of the E gene is lower than that of the E subregion. Some studies have shown that most of the mice are expressed in the MHC II I-Ab gene, and this is the essential gene for the survival of the species. Therefore, this experiment is to choose the relatively low polymorphic A subregion as the target area and through the constant gene fragment in the A B subregion. Detection of segment, analysis and study of different stress sources
2.
The difference of immune response between the body and its related mechanism is attempted.
A subacute restraint stress model was established in mice, which is different from acute stress.
Mental stress model) and subacute heat stress model (prone to physical stress model).
On day 3,10,16, mice were immunized with ovalbumin antigen and died in two groups of mice.
At the time of 1/3, blood was collected from the caudal vein, and blood was collected once a day for 7 days at the time of death 2/3.
Under different conditions, enzyme linked immunosorbent assay (ELISA) was used to treat different stress models.
Dynamic detection of antibody level in mice was used to analyze subacute psychological stress and subacute body.
The influence of stress on immune response was established, and a 9 day subacute stress animal model was established.
The cortisol levels in the blood of mice in different stress groups and control groups were quantitatively detected.
Fluorescein (FITC) labeled H-2IAb monoclonal antibodies were analyzed by flow cytometry.
The expression of H-2IAb on lymphocyte surface of stress mice was quantitatively detected, and one-step method was adopted.
The expression level of H-2IAb gene was detected by RT-PCR, and subacute in 9 days on day 1,5.
The stress animal model mice were immunized with ovalbumin antigen, and the body weight of mice was measured.
The relationship between the weight change rate and the immune index was analyzed.
The results showed that there were significant differences in the effects of two subacute stress on humoral immune response.
(P0.05) in the 7 day test, the level of anti ovalbumin antibody (U) in the restraint stress group was low.
In the heat stress group, cortisol test results showed that the content of cortisol in blood of two stress group mice
(nmol/L) were higher than those in the normal control group, but there was no statistical difference (restraint stress group 5.57 + 1.75).
The heat stress group was 4.20 + 1.36, the control group was 3.91 + 0.99, P0.05).
The results showed that the expression of IAb on lymphocyte surface of different stress groups was significantly different (Mean.).
The restraint stress group was significantly lower than that of the heat stress group (2.86 + 1.76,6.83 + 2.32, P0.05).
RT-PCR electrophoresis showed that H-2 IAb gene was detected in all mice, which was visible to the naked eye.
The intensity difference was also correlated with the result of flow cytometry (correlation coefficient r=0.649).
The accuracy of monoclonal antibodies in flow cytometry was demonstrated, and the immunization index (U) occurred in mice.
At the same time (31.89 + 28.92) in the restraint stress group, 76.06 + 72.71 in the heat stress group, the control group.
87.44 + 57.35). There was a significant difference in the weight change rate between the three groups (restraint stress group).
-0.09 + 0.06, heat stress group -0.01 + 0.05, control group 0.11 + 0.06), and ruled out immunity.
The correlation between the index and the rate of body weight change.
The following conclusions are reached through analysis: 1.
The animal model is a subacute restraint stress model which tends to psychological stress and tends to the body.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392

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