本文選題：應(yīng)激 + 免疫應(yīng)答�。� 參考：《大連醫(yī)科大學(xué)》2005年碩士論文

【摘要】：應(yīng)激(stress)反應(yīng)是人類日常生活中不可避免的,創(chuàng)傷、高熱、工作壓力、應(yīng)試等均是其常見的應(yīng)激源。一般將應(yīng)激定義為:機(jī)體對不協(xié)調(diào)狀態(tài)或內(nèi)環(huán)境穩(wěn)定受到威脅時所做出的反應(yīng)。亞急性應(yīng)激或慢性應(yīng)激,一般指超過 24 小時的持續(xù)或反復(fù)應(yīng)激,主要伴有神經(jīng)、內(nèi)分泌、免疫和行為等變化。處于應(yīng)激狀態(tài)的軀體可產(chǎn)生一系列的生理、心理、內(nèi)分泌、神經(jīng)生化及免疫功能等方面的改變,這是機(jī)體受到應(yīng)激而做出的適應(yīng)性全身應(yīng)答,借以維持機(jī)體內(nèi)環(huán)境的相對穩(wěn)定。同時,隨著社會的發(fā)展,人們意識到亞急性應(yīng)激比急性應(yīng)激更能嚴(yán)重影響人類的身心健康。所以,亞急性應(yīng)激在疾病的發(fā)生、發(fā)展中所起的作用也越來越受到關(guān)注。但國內(nèi)有關(guān)這方面的研究尚處于剛剛起步階段,尤其在不同應(yīng)激源對機(jī)體免疫系統(tǒng)是否有影響、有何影響的方面還沒有系統(tǒng)詳盡的研究報道。 已有研究表明,位于小鼠的 17 號染色體上的 H-2 復(fù)合體(Histocompatibility 2 complex, H-2 complex),即主要組織相容性復(fù)合體(Major Histocompatibility Complex, MHC),是多態(tài)性最豐富的基因系統(tǒng)。H-2 復(fù)合體 I 區(qū)中分立的基因編碼小鼠 II 類分子的α和β鏈。其中最主要的為 A 基因和 E 基因。A 亞區(qū)的基因多態(tài)性低于 E 亞區(qū)。有研究表明,絕大多數(shù)種系小鼠均表達(dá) MHC II 類 I-Ab 基因,此為種系生存的必需基因。因此,本實驗即選用多態(tài)性相對低的 A 亞區(qū)為目標(biāo)區(qū),通過對 A b 亞區(qū)內(nèi)恒定基因片段的檢測,分析研究不同應(yīng)激源對 ·2· 機(jī)體免疫反應(yīng)影響的差異性,并試圖找出其相關(guān)機(jī)制。 本文建立了不同于急性應(yīng)激的小鼠亞急性束縛應(yīng)激模型(傾向于 心理應(yīng)激模型)與亞急性熱應(yīng)激模型(傾向于軀體應(yīng)激模型),并于造 模第 3、10、16 天對小鼠進(jìn)行卵清蛋白抗原免疫,于兩組小鼠各死亡 1/3 時進(jìn)行尾靜脈采血,至死亡 2/3 時止,每天采血一次,共計 7 天。 在可比的條件下用酶聯(lián)免疫吸附實驗(ELISA)對不同性質(zhì)應(yīng)激模型組 小鼠抗體激發(fā)水平進(jìn)行動態(tài)檢測,分析亞急性心理應(yīng)激與亞急性軀體 應(yīng)激對免疫應(yīng)答的影響;同時建立了小鼠 9 天亞急性應(yīng)激動物模型, 對不同應(yīng)激組及對照組小鼠血液中皮質(zhì)醇進(jìn)行定量檢測;借助異硫氰 熒光素(FITC)標(biāo)記的 H-2IAb 單克隆抗體,運(yùn)用流式細(xì)胞術(shù),對不同 應(yīng)激小鼠淋巴細(xì)胞表面 H-2IAb 的表達(dá)進(jìn)行定量檢測;同時采用一步法 RT-PCR 的方法檢測 H-2IAb 基因表達(dá)水平;于第 1、5 天對 9 天亞急性 應(yīng)激動物模型小鼠進(jìn)行卵清蛋白抗原免疫,通過測定小鼠的體重變化, 分析體重變化率與免疫指標(biāo)的關(guān)系。 結(jié)果顯示兩種亞急性應(yīng)激對體液免疫應(yīng)答的影響存在顯著性差異 (p0.05),7 天的檢測中,束縛應(yīng)激組抗卵清蛋白抗體水平(U)低 于熱應(yīng)激組;皮質(zhì)醇檢測結(jié)果顯示兩應(yīng)激組小鼠血液中皮質(zhì)醇含量 (nmol/L)均高于正常對照組,但無統(tǒng)計學(xué)差異(束縛應(yīng)激組 5.57±1.75, 熱應(yīng)激組 4.20±1.36,對照組 3.91±0.99,p0.05);流式細(xì)胞儀檢測結(jié) 果顯示不同應(yīng)激組小鼠淋巴細(xì)胞表面 IAb 的表達(dá)(Mean.)有顯著性差 異,束縛應(yīng)激組顯著低于熱應(yīng)激組(2.86±1.76,6.83±2.32,p0.05); RT-PCR 電泳結(jié)果顯示所有小鼠均檢測出 H-2 IAb 基因,有肉眼可見的 強(qiáng)度區(qū)別,同時,與流式細(xì)胞儀檢測結(jié)果相關(guān)(相關(guān)系數(shù) r=0.649), 證明了流式細(xì)胞儀中單克隆抗體的準(zhǔn)確性;在小鼠免疫指標(biāo)(U)發(fā)生 變化的同時(束縛應(yīng)激組 31.89±28.92,熱應(yīng)激組 76.06±72.71,對照組 87.44±57.35),三組小鼠間體重變化率存在顯著性差異(束縛應(yīng)激組 -0.09±0.06,熱應(yīng)激組-0.01±0.05,對照組 0.11±0.06),同時排除了免疫 指標(biāo)與體重變化率之間的相關(guān)性。 通過分析得出以下結(jié)論:1、本實驗成功建立了不同性質(zhì)亞急性應(yīng) 激動物模型,即傾向于心理應(yīng)激的亞急性束縛應(yīng)激模型與傾向于軀體
[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.
【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2005
【分類號】：R392

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