本文關(guān)鍵詞：個(gè)體特質(zhì)與生物標(biāo)志物應(yīng)激反應(yīng)模式相關(guān)性研究　出處：《東南大學(xué)》2016年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 應(yīng)激 人格 生物標(biāo)志物 唾液α-洳淀粉酶 化學(xué)發(fā)光 免疫 運(yùn)動(dòng)干預(yù)

【摘要】：應(yīng)激是機(jī)體對(duì)環(huán)境刺激具有非特異性的生物學(xué)反應(yīng)現(xiàn)象。應(yīng)激對(duì)健康、行為及工作效率的影響已成為國際上重要的職業(yè)衛(wèi)生問題之一,所以評(píng)價(jià)應(yīng)激程度具有非常重要的意義。各種內(nèi)外環(huán)境因素及社會(huì)、心理因素刺激均可以使人體或動(dòng)物產(chǎn)生應(yīng)激反應(yīng),這種反應(yīng)會(huì)在不同程度上引起認(rèn)知能力的改變。有文獻(xiàn)研究表明,情緒穩(wěn)定性作為一種人格特征,存在明顯的個(gè)體差異,同時(shí)對(duì)個(gè)體具有相對(duì)穩(wěn)定性。這種穩(wěn)定性既有認(rèn)知評(píng)價(jià)、人格特征、氣質(zhì)類型等個(gè)性因素,也有生理學(xué)基礎(chǔ)。隨著近代生命科學(xué)的發(fā)展,研究工作者對(duì)應(yīng)激認(rèn)識(shí)從早期的現(xiàn)象觀察和描述,逐漸發(fā)展為從“心理-神經(jīng)內(nèi)分泌-行為”多學(xué)科多角度,更加系統(tǒng)全面地揭示應(yīng)激的生理與心理響應(yīng)特征,及應(yīng)激對(duì)大腦、心理與行為活動(dòng)影響的深層機(jī)制。應(yīng)激生物標(biāo)志物檢測研究一直是生理學(xué)、醫(yī)學(xué)、運(yùn)動(dòng)學(xué)、心理學(xué)研究等所關(guān)注的重點(diǎn)。深入研究基于應(yīng)激標(biāo)志物的評(píng)估方法及應(yīng)激響應(yīng)規(guī)律,對(duì)預(yù)防各類應(yīng)激損傷有重要意義。目前,應(yīng)激評(píng)估方法主要有問卷調(diào)查法、行為觀察法、電生理、生化及腦成像檢測法等多種方法。其中,基于生化檢測的評(píng)估方法具有客觀、準(zhǔn)確的優(yōu)點(diǎn)。但是,對(duì)生物樣本中的神經(jīng)活性物質(zhì)進(jìn)行定量分析需要非常靈敏、精密的分析方法,這已成為相關(guān)研究順利開展的關(guān)鍵環(huán)節(jié)。本文選取無創(chuàng)樣本唾液,并以唾液α一淀粉酶和皮質(zhì)醇等為目標(biāo)物,深入探究其作為應(yīng)激標(biāo)志物在認(rèn)知神經(jīng)科學(xué)等相關(guān)研究中的可靠性和有效性。本文主要開展了以下六個(gè)主要內(nèi)容的研究工作：(1)應(yīng)激研究技術(shù)平臺(tái)的建立及優(yōu)化；(2)多參數(shù)評(píng)價(jià)應(yīng)激對(duì)小鼠腦與行為的影響；(3)運(yùn)動(dòng)對(duì)應(yīng)激標(biāo)志物的影響初步研究；(4)多參數(shù)評(píng)價(jià)人體心理應(yīng)激研究；(5)個(gè)體特質(zhì)在心理應(yīng)激中的調(diào)節(jié)作用；(6)應(yīng)激對(duì)注意調(diào)控能力的影響。具體工作如下：一、應(yīng)激研究技術(shù)平臺(tái)的建立及優(yōu)化(1)生物標(biāo)志物快速檢測技術(shù)的構(gòu)建及優(yōu)化①化學(xué)發(fā)光免疫法快速測定前白蛋白方法的建立：將聚乙烯亞胺(PEI)修飾的磁顆粒(MBs)通過戊二醛偶聯(lián)抗體(Nb1),所形成的免疫磁珠(MB/PEI/Nb1)能夠特異的結(jié)合前白蛋白(PA)形成免疫復(fù)合物。該復(fù)合物可與包被有辣根過氧化物酶(HRP)和檢測抗體(Nb2)的SiO2納米粒子(HRP-SiO2-Nb2)形成免疫夾心復(fù)合物(MB/PEI/Nb1-PA-Nb2/SiO2-HRP)。復(fù)合物中的HRP催化魯米諾、對(duì)碘苯酚(PIP)及H2O2反應(yīng),提高化學(xué)發(fā)光(CL)信號(hào)。該方法的檢測限為0.01μgL-1,線性范圍0.05到1000μg L-1,完成整個(gè)操作只需40分鐘。②六種單胺類神經(jīng)遞質(zhì)的高效液相色譜-熒光檢測法的優(yōu)化：6種目標(biāo)物分別為去甲腎上腺素(NE)、多巴胺(DA)、3,4二羧基苯乙酸(DOPAC)、3-甲基-4-羥基苯乙二醇(MHPG)、5-羥色胺、5-羥吲哚乙酸(5-HIAA),采用CosmosilC18 (4.6x250 mm,5μm)色譜柱,流動(dòng)相：0.1mol/L的醋酸鈉溶液：甲醇(體積比5：95),pH為6,其中含0.2mmol/L乙二胺四乙酸二鈉；流速2.0 mL/min;在2-300pmoL范圍內(nèi),目標(biāo)物具有較好的線性關(guān)系(0.999),每種神經(jīng)遞質(zhì)的回收率為95%-102%。優(yōu)化后的方法適用于腦組織不同區(qū)域的單胺類神經(jīng)遞質(zhì)檢測。③唾液α-淀粉酶(sAA)流動(dòng)注射分析方法優(yōu)化：在本組前期研究的基礎(chǔ)上進(jìn)行進(jìn)一步優(yōu)化條件,建立了更加快速、簡便的sAA流動(dòng)注射光度測定法。該方法測定一個(gè)樣本的周期僅為34秒,可以實(shí)現(xiàn)自動(dòng)化操作,精密度高,避免人工操作上時(shí)間控制不當(dāng)所導(dǎo)致的測定差異大的缺點(diǎn)。④其他檢測方法的建立：基因檢測方法、近紅外腦成像檢測技術(shù)。(2)應(yīng)激模型的構(gòu)建及優(yōu)化應(yīng)激模型是研究應(yīng)激與神經(jīng)內(nèi)分泌、行為關(guān)聯(lián)性的重要手段。因?yàn)榇蟛糠盅芯慷际墙?dòng)物模型,因此人類數(shù)據(jù)有限,但嚙齒類動(dòng)物和非人靈長類的一些研究數(shù)據(jù),可以應(yīng)用于人體模型。通過文獻(xiàn)總結(jié),我們分別建立了針對(duì)動(dòng)物和人體應(yīng)激相關(guān)的模型,包括小鼠應(yīng)激模型和人體應(yīng)激模型。(3)應(yīng)激評(píng)價(jià)方法的構(gòu)建為了考察應(yīng)激對(duì)機(jī)體的影響,本課題選擇了多種實(shí)驗(yàn)范式以測評(píng)動(dòng)物及人體行為。針對(duì)小鼠的有：高架十字迷宮、攝食抑制行為、強(qiáng)迫游泳實(shí)驗(yàn)、社交行為實(shí)驗(yàn)；針對(duì)人體的有：Stroop實(shí)驗(yàn)、心算評(píng)價(jià)、眼動(dòng)實(shí)驗(yàn)等。二、多參數(shù)評(píng)價(jià)應(yīng)激對(duì)小鼠腦及行為的影響束縛應(yīng)激是研究早期事件對(duì)行為改變廣泛應(yīng)用的模型之一。但是束縛應(yīng)激能否引起社交焦慮行為,運(yùn)動(dòng)是否能夠起到有效干預(yù)作用尚存有爭議。因此,我們對(duì)1月齡小鼠分別進(jìn)行了每天兩小時(shí),共兩周的束縛應(yīng)激試驗(yàn)研究。結(jié)果表明,相對(duì)于正�？刂平M,束縛應(yīng)激小鼠(出生后28天-37天)焦慮和抑郁行為明顯提高。此外,束縛應(yīng)激小鼠行為改變還伴隨其海馬HDACsmRNA表達(dá)和不同腦區(qū)神經(jīng)遞質(zhì)水平的改變。有氧鍛煉有效的緩解了束縛應(yīng)激導(dǎo)致的單胺類神經(jīng)遞質(zhì)、HDACs mRN A表達(dá)和多種行為的改變。研究結(jié)果說明束縛應(yīng)激會(huì)導(dǎo)致社交及情緒相關(guān)行為的改變,而運(yùn)動(dòng)干預(yù)會(huì)起到緩解作用。三、運(yùn)動(dòng)對(duì)應(yīng)激標(biāo)志物的影響初步研究本文主要探討兒童在運(yùn)動(dòng)應(yīng)激條件下,sAA活力指標(biāo)的變化規(guī)律,考察其作為一種客觀的評(píng)價(jià)運(yùn)動(dòng)應(yīng)激的生物學(xué)參數(shù)的可能性。被試為120名小學(xué)生。在運(yùn)動(dòng)應(yīng)激條件下,采集他們測試前30分鐘、測試結(jié)束后即刻及測試結(jié)束休息30分鐘后的唾液樣品。利用流動(dòng)注射法測定sAA的濃度。本研究的結(jié)果顯示,被試的sAA水平在運(yùn)動(dòng)應(yīng)激的不同階段之間無顯著性差異,但在不同年級(jí)分組之間明顯存在個(gè)體差異；結(jié)合問卷分析,我們推測sAA活力水平可能受到人格對(duì)應(yīng)激的評(píng)估和調(diào)節(jié)作用。流動(dòng)注射快速測定sAA的方法在客觀評(píng)價(jià)運(yùn)動(dòng)應(yīng)激方面有潛在應(yīng)用價(jià)值。四、多參數(shù)評(píng)價(jià)人體心理應(yīng)激研究探討在心理應(yīng)激壓力下,sAA指標(biāo)與心理應(yīng)激的相關(guān)性,考察其作為一種客觀的評(píng)價(jià)心理應(yīng)激的生物學(xué)參數(shù)的可能性。被試為44名研究生(男女生各22名),年齡為23-27歲。在Stroop測試、心算測試心理應(yīng)激條件下,采集他們測試前10分鐘、測試結(jié)束時(shí)以及測試結(jié)束休息10分鐘后的唾液樣品。利用流動(dòng)注射法測定sAA的濃度。結(jié)果顯示被試的sAA水平在心理應(yīng)激實(shí)驗(yàn)時(shí)明顯變化,也表明流動(dòng)注射快速測定sAA的方法可用于客觀評(píng)價(jià)心理應(yīng)激。五、個(gè)體特質(zhì)在心理應(yīng)激研究中的調(diào)節(jié)作用為了驗(yàn)證TPQ問卷中的3個(gè)人格維度是否與應(yīng)激的神經(jīng)內(nèi)分泌反應(yīng)相關(guān),我們招募了64名健康本科學(xué)生,通過填寫TPQ問卷,將全部被試分別分為獵奇(NS)、危害避免(HA)及獎(jiǎng)賞效應(yīng)(RD)值高低不同的兩組。所有被試均參加Stroop聯(lián)合心算的應(yīng)激測試并測定心率變異性和sAA指標(biāo),統(tǒng)計(jì)分析分析比較組間差異。結(jié)果表明：①高NS組Stroop完成數(shù)量顯著高于低NS組；高NS、HA組Stroop測試平均反應(yīng)時(shí)顯著高于低NS、HA組。②Stroop測試后sAA活力水平顯著高于測試前的安靜值；心算后高HA組的sAA水平顯著高于低HA組。③心率變異性中的Stroo P前均一化低頻成分Stroop前(LFnul),均一化化高頻成分(HFnul),低頻／高頻(LF/HF1), srtoop后相鄰RR間期差值的均方根(RMSSD2),均一化化低頻(HF2)、心算后即刻高頻成分(HF3),相鄰正常R-R間期差值均方根(RMSSD3)等指標(biāo)高NS、RD組顯著高于低NS、RD組。結(jié)果顯示不同人格維度與sAA活力和心率變異性參數(shù)間存在相關(guān)性,對(duì)人格的生物基礎(chǔ)提供了理論支持。結(jié)果說明TPQ人格中的獵奇和獎(jiǎng)賞維度可能是影響應(yīng)激響應(yīng)特征的重要調(diào)節(jié)因子。六、應(yīng)激對(duì)注意認(rèn)知調(diào)節(jié)能力的影響視覺注意的認(rèn)知調(diào)控常用于運(yùn)動(dòng)員技戰(zhàn)術(shù)的日常訓(xùn)練,但在比賽中采用這種策略卻很難奏效。本研究深入探索了高水平重劍運(yùn)動(dòng)員和普通大學(xué)生在面對(duì)模擬對(duì)手進(jìn)攻時(shí)視覺注意線索的變化特點(diǎn)。學(xué)生組隨機(jī)分為應(yīng)激組和控制組,應(yīng)激組接受冷水刺激。刺激后,檢測所有被試對(duì)模擬對(duì)手進(jìn)攻的視覺注意眼動(dòng)規(guī)律,并檢測sAA及唾液皮質(zhì)醇指標(biāo)。結(jié)果發(fā)現(xiàn),專業(yè)運(yùn)動(dòng)員有更高效的首注視點(diǎn)注意效率。應(yīng)激降低了普通大學(xué)生視覺注意的認(rèn)知調(diào)節(jié)能力。唾液應(yīng)激標(biāo)志物可以作為評(píng)價(jià)不同特質(zhì)個(gè)體視覺注意控制能力的潛在指標(biāo),評(píng)價(jià)注意認(rèn)知調(diào)節(jié)水平。
[Abstract]:Stress is the body of environmental stimulation biological reaction phenomenon is nonspecific. Effects of stress on health, behavior and work efficiency has become one of the important international occupation health problems, so the evaluation of the degree of stress has very important significance. All kinds of internal and external environment factors and social factors, psychological stimulation can make human or animal stress reaction this reaction, will cause the changes of cognitive ability in different extent. A literature research shows that emotional stability as a personality characteristic, the obvious individual differences, also has the relative stability of the individual. The stability of both cognitive evaluation, personality, temperament and personality factors, but also the basis of Physiology. With the development of in modern life science, researchers stress understanding from the phenomenon observed and described earlier, gradually developed from "Psycho neuroendocrine "Multi-disciplinary perspective, more systematically reveal the physiological characteristics and the psychological stress response, and stress on the brain, deep influence mechanism of psychological and behavioral activities. The stress of biomarker detection has been physiology, medicine, kinematics, psychology and other key concern. Further research on evaluation method of stress markers and based on the stress response, has important significance for the prevention of various types of stress damage. At present, stress evaluation methods mainly include questionnaire, behavioral observation, electrophysiological, biochemical methods and brain imaging detection method. The evaluation method of biochemical detection based on an objective, accurate. However, quantitative analysis need to be very sensitive to neural active substances in biological samples, sophisticated analytical methods, it has become a key link in related research carried out smoothly. This paper selects noninvasive saliva samples, And to the alpha amylase and salivary cortisol as a target to explore the reliability as stress markers in related research in cognitive neuroscience and effectiveness. The research work of this paper mainly includes the following six main contents: (1) establish and optimize the stress of technology platform; (2) effect the parameters of stress evaluation on brain and behavior; (3) a preliminary study on the influence of exercise stress; (4) to study the human psychological stress evaluation parameters; (5) individual role in mental stress; (6) pay attention to the impact of stress on the regulatory capacity. The specific work is as follows: 1. Establish and optimize the stress research platform (1) biomarkers for rapid detection of prealbumin determination techniques of construction and optimization of the chemiluminescence immunoassay: polyethyleneimine (PEI) modified magnetic particles (MBs) by Glutaraldehyde coupling antibody (Nb1), formed by immunomagnetic beads (MB/PEI/Nb1) can be combined with specific prealbumin (PA) formation of immune complexes. The complexes can be coated with horseradish peroxidase (HRP) and antibody (Nb2) SiO2 nanoparticles (HRP-SiO2-Nb2) formed a sandwich assay (MB/PEI/Nb1-PA-Nb2/SiO2-HRP). HRP catalyzed Lumino in the compound of iodine, phenol (PIP) and H2O2 reaction, enhance the chemiluminescence (CL) signal. The detection limit is 0.01 gL-1, the linear range of 0.05 to 1000 g L-1, the entire operation was completed in just 40 minutes. The optimization of high performance liquid phase six monoamine neurotransmitters HPLC - fluorescence detection method: 6 targets were norepinephrine (NE), dopamine (DA), 3,4 (DOPAC) two carboxyl phenyl acetic acid, 3- methyl -4- hydroxy phenylglycol (MHPG), 5- serotonin, 5- hydroxyindoleacetic acid (5-HIAA), using CosmosilC18 (4.6x250 mm, 5 M) column, mobile phase: 0.1mol/L sodium acetate solution: methanol (volume ratio 5:95), pH was 6, including two 0.2mmol/L EDTA sodium 2 mL/min; flow rate; in the range of 2-300pmoL, the object has a good linear relationship (0.999), the recovery rate for each neurotransmitter 95%-102%. optimization method after the application of monoamine neurotransmitters in different brain region detection. The salivary amylase (sAA) optimization method for flow injection: to optimize the conditions based on the previous research, the establishment of a more rapid, sAA flow injection spectrophotometric method for the determination of the simple cycle. The method for the determination of a sample only 34 seconds, can realize automatic operation, high precision, avoid the manual operation on time control caused by improper determination of differences. The establishing of defect detection method: gene detection method, near infrared brain As the detection technology. (2) stress model and optimization model of stress and stress is an important means of neuroendocrine, behavior related. Because most of the research is to establish the animal model of the human data is limited, but animal rodents and non-human primates some research data can be applied to the human body through the model. The literature review, we developed for animal and human stress related models, including the model and the stress model of stress in mice. (3) stress evaluation method constructed in order to investigate the effects of stress on the body, this paper selects the various experimental paradigms to assess animal and human behavior. In mice: the elevated plus maze, inhibition of feeding behavior, forced swimming test, social behavior experiment; in view of the human body are: Stroop experiment, mental evaluation, eye movement experiment. Two, multi parameter evaluation of mouse brain and stress The effect of restraint stress is one of the early events of the widely used model of behavior change. But the restraint stress can cause anxiety and social behavior, whether exercise can play a role in effective intervention remains controversial. Therefore, we are on January old mice two hours a day, experimental study on restraint stress for two weeks. The results showed that and compared with the normal control group, restraint stress mice (postnatal 28 days -37 days) anxiety and depression behavior improved significantly. In addition, restraint stress mice behavior change with the change of the expression of HDACsmRNA in hippocampus and brain neurotransmitter levels. Aerobic exercise can effectively relieve the monoamine neurotransmitters in restraint stress caused by HDACs mRN the expression of A and a variety of behavioral changes. Research results indicate that restraint stress can lead to social and emotional behavior change, and exercise intervention will play a role in mitigation. Three, the preliminary study on the influence of this paper mainly discusses the children in the exercise stress conditions exercise stress signs, changes of sAA activity index, investigate the possibility as biological parameters of an objective evaluation of exercise stress. The subjects were 120 students. In the exercise stress conditions, they collected 30 minutes before the test. After the end of the test and at the end of the test after 30 minutes of rest of the saliva samples. SAA concentrations were determined by flow injection method. The results of this study showed that there was no significant difference between subjects sAA levels at different stages of exercise stress, but there are obvious individual differences among different grade groups; combined with the analysis of the questionnaires, we speculate that the level of sAA activity may be evaluated on stress and personality regulation. Rapid determination method of sAA flow injection has the potential application value in the evaluation of exercise stress. More than four. The study of human psychological stress parameters to evaluate the in psychological stress, correlation between sAA index and psychological stress, to investigate the possibility as biological parameters of an objective evaluation of the psychological stress. The subjects were 44 students (male and female 22), aged 23-27 years old. In the Stroop test, the psychological stress condition under the mental test, they collected 10 minutes before the test, the end of the test and the test ended after 10 minutes of rest of the saliva samples. SAA concentrations were determined by flow injection method. The results showed that the level of sAA was changed in psychological stress experiment, also shows that the method of quick determination of sAA flow injection can be used to evaluate the psychological stress. Five, regulation of individual characteristics in the study of psychological stress in order to verify whether the 3 personality dimensions of TPQ questionnaire related to the neuroendocrine response to stress, we recruited 64 healthy the The students by filling out TPQ questionnaire, all subjects were divided into Adventures (NS) (HA), to avoid harm and reward effect (RD) values of different two groups. All subjects are in Stroop United stress test and determination of heart rate variability and sAA index, statistical analysis differences between groups. The results showed: high NS groups of Stroop number was significantly higher than that of low NS group; high NS, HA group Stroop test average reaction time was significantly higher than that of low NS, HA group. SAA activity level of Stroop was significantly higher than that before the test after the test value after sAA quiet; mental water high HA group was significantly higher than that of low HA group. The heart rate variability in Stroo P before homogenization of low-frequency components before Stroop (LFnul), the homogenization of high frequency components (HFnul), low frequency / high frequency (LF/HF1), srtoop after the adjacent RR interval difference of root mean square (RMSSD2), the homogenization of the low frequency (HF2), immediately after the high frequency components (mental arithmetic HF3), adjacent Normal R-R interval difference of root mean square (RMSSD3) and other indicators of RD group was significantly higher than that of high NS, low NS, RD group. The results showed that the correlation between different personality dimensions and sAA activity and heart rate variability among the parameters, provides a theoretical support for the biological basis of personality. The results show that the TPQ personality curiosity and reward dimension may be the important regulatory factors affecting stress response characteristics. Six, stress on the attention of visual attention cognition regulation used in the daily training of athletes'tactics influence cognitive regulation ability, but in the game using this strategy is hard work. This study explores the deep into the high level epee athletes and college students in the face of simulated opponents at visual attention characteristics of clues. The students were randomly divided into stress group and control group, stress group received cold stimulation. After stimulation, detection of all subjects attack on the opponent's attention to eye visual simulation The dynamic law, and detection of sAA and salivary cortisol index. The results showed that professional athletes are the first focus the attention of more efficient efficiency. Stress reduces the visual attention of ordinary college students cognitive regulation ability. Saliva stress markers can be used as a potential indicator control capability evaluation of different individual characteristics of visual attention, pay attention to adjust the level of cognitive evaluation.

【學(xué)位授予單位】：東南大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R33

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