本文選題：心理應(yīng)激哮喘大鼠模型 + 肝主疏泄�。� 參考：《北京中醫(yī)藥大學(xué)》2017年博士論文

【摘要】：研究目的哮喘是典型的心身疾病,心理應(yīng)激在疾病過(guò)程中起著重要作用。既往研究顯示,情志可引起神經(jīng)-內(nèi)分泌-免疫失衡,HPA軸是其主要調(diào)節(jié)途徑之一。心理應(yīng)激條件下,哮喘氣道炎癥明顯增強(qiáng),對(duì)激素反應(yīng)性下降;其作用機(jī)制可能與糖皮質(zhì)激素受體表達(dá)及功能下調(diào),激素敏感性下降有關(guān)。本研究模擬臨床情志致病特點(diǎn),建立心理應(yīng)激哮喘大鼠模型,通過(guò)相關(guān)指標(biāo)檢測(cè),揭示心理應(yīng)激哮喘大鼠HPA軸功能變化情況及其與氣道炎癥反應(yīng)的相關(guān)性;并通過(guò)哮喘寧顆粒對(duì)心理應(yīng)激哮喘大鼠進(jìn)行干預(yù),觀察模型大鼠宏觀表征、行為學(xué)以及血清、灌洗液、肺組織相關(guān)指標(biāo)變化,探索調(diào)肝理肺法對(duì)心理應(yīng)激哮喘大鼠HPA軸及氣道炎癥的調(diào)控作用,闡明調(diào)肝理肺法的作用機(jī)理和作用環(huán)節(jié)。研究方法本研究在前期工作基礎(chǔ)上,將雄性SD大鼠隨機(jī)分為正常組、應(yīng)激組、哮喘組、應(yīng)激哮喘組、西藥組和中藥組。并模擬臨床情志致病特點(diǎn),采用卵蛋白混合致敏聯(lián)合束縛制動(dòng)的方法,建立心理應(yīng)激哮喘大鼠模型,從宏觀表征、行為學(xué)、肺通氣功能等客觀指標(biāo)進(jìn)行模型評(píng)價(jià);并通過(guò)檢測(cè)血清、灌洗液內(nèi)CRH、ACTH、CORT水平,肺組織GR表達(dá)及氣道炎癥因子含量,深入研究模型大鼠HPA軸功能的變化及其與氣道炎癥反應(yīng)的相關(guān)性。并用哮喘寧顆粒對(duì)心理應(yīng)激哮喘大鼠進(jìn)行干預(yù),與其余各組相比較,觀察血清及BALF、肺組織中相關(guān)指標(biāo)濃度、陽(yáng)性細(xì)胞表達(dá)及蛋白表達(dá)的變化情況,研究調(diào)肝理肺法對(duì)心理應(yīng)激哮喘大鼠HPA軸功能及免疫炎癥失衡的調(diào)節(jié)作用。研究結(jié)果1)采用OVA混合致敏聯(lián)合束縛制動(dòng)成功建立心理應(yīng)激哮喘大鼠模型。結(jié)果顯示應(yīng)激哮喘組大鼠表現(xiàn)為興趣減退、毛發(fā)晦暗無(wú)光澤、便溏、飲食量減少、體重增加緩慢;激發(fā)過(guò)程中有抓咬口鼻、體腹部皮膚,嗆咳、噴嚏及喉中哮鳴音等表現(xiàn)。與正常組相比較,應(yīng)激哮喘組水平運(yùn)動(dòng)、垂直運(yùn)動(dòng)得分明顯下降(P0.05,P0.01);蔗糖水偏好程度、懸吊實(shí)驗(yàn)掙扎次數(shù)下降(P0.01);懸吊實(shí)驗(yàn)中不動(dòng)時(shí)間明顯增加(P0.01);肺通氣功能 PEF、FEF25-75%、FEV0.3/FVC 下降(P0.05,P0.01,P0.01),氣道阻力Ri、Re明顯升高(P0.05,P0.01);靜脈血嗜酸粒細(xì)胞升高(P0.01);肺組織炎癥細(xì)胞浸潤(rùn)明顯。與應(yīng)激組或哮喘組相比,應(yīng)激性哮喘組的癥狀、體征和相關(guān)指標(biāo)均明顯惡化。2)與正常組比較,應(yīng)激哮喘組大鼠血清、BALF內(nèi)CRH、ACTH、CORT及IgE、IL-4、IL-17含量明顯增高(P0.01),IFN-γ含量明顯下降(P0.01)。肺組織中GR陽(yáng)性表達(dá)及含量明顯下降(P0.01);而IL-4、IL-17陽(yáng)性細(xì)胞表達(dá)及含量增高明顯(P0.01);IFN-γ陽(yáng)性細(xì)胞表達(dá)明顯減低(P0.01),而肺組織IFN-γ含量輕度減低,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。與應(yīng)激組或哮喘組相比,應(yīng)激性哮喘組的HPA軸及氣道炎癥相關(guān)指標(biāo)均有不同程度加重。相關(guān)性研究顯示,CORT與IL-4、IL-17、IgE呈正相關(guān),與IFN-y呈負(fù)相關(guān);而肺組織中GR水平與之相反,即與IFN-y含量呈正相關(guān),而與IL-4、IL-17等呈負(fù)相關(guān)。而CORT含量與心理應(yīng)激哮喘大鼠行為學(xué)指標(biāo)相關(guān)性研究發(fā)現(xiàn),CORT含量與糖水消耗量、糖水比例等呈負(fù)相關(guān),而肺組織GR含量與糖水消耗量、糖水比例、翻動(dòng)次數(shù)及水平垂直運(yùn)動(dòng)呈正相關(guān);但與靜止時(shí)間呈負(fù)相關(guān),相關(guān)性較弱。3)哮喘寧顆粒治療后,中藥組大鼠前期活動(dòng)減少,反應(yīng)遲緩,毛發(fā)晦暗,后期情緒改善,較為活躍,飲食量增加,對(duì)周?chē)挛锱d趣增高;激發(fā)過(guò)程中呼吸頻率增快,腹肌收縮加深、加快,時(shí)有搔抓頭面、體腹部毛發(fā)。與模型組相比較,中藥組大鼠水平運(yùn)動(dòng)、垂直運(yùn)動(dòng)得分明顯增加(P0.01);蔗糖水偏好程度、懸吊實(shí)驗(yàn)掙扎次數(shù)明顯增高(P0.01,P0.01);懸吊實(shí)驗(yàn)不動(dòng)時(shí)間明顯降低(P0.01);肺通氣功能中藥組大鼠PEF、FEF25-75%、FEV0.3/FVC輕度增高;而Ri、Re減低,差異有顯著統(tǒng)計(jì)意義(P0.01);靜脈血嗜酸粒細(xì)胞明顯降低(P0.01);肺組織中炎癥細(xì)胞浸潤(rùn)明顯減少。中藥組大鼠血清、BALF內(nèi)CRH、ACTH、CORT含量較應(yīng)激哮喘組明顯下降(P0.01),而肺組織GR陽(yáng)性表達(dá)明顯增高(P0.01),其含量輕度升高,差異無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)。而血清、BALF及支氣管肺組織IL-4、IL-17含量及陽(yáng)性細(xì)胞表達(dá)明顯下降(P0.01);肺組織IFN-γ陽(yáng)性細(xì)胞表達(dá)明顯降低(P0.01),而含量略有降低,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。相關(guān)性研究結(jié)果顯示CORT含量與IFN-γ呈負(fù)相關(guān),而與IgE、IL-4、IL-17等炎癥指標(biāo)呈正相關(guān);而肺組織GR含量與IFN-γ水平呈正相關(guān),而與IL-4、IL-17等炎癥指標(biāo)呈負(fù)相關(guān)。而CORT含量與大鼠行為學(xué)相關(guān)性研究發(fā)現(xiàn),CORT含量與糖水消耗量、糖水比例、翻動(dòng)次數(shù)及垂直運(yùn)動(dòng)次數(shù)呈負(fù)相關(guān);而與靜止時(shí)間呈正相關(guān),但相關(guān)性均較弱,無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)。研究結(jié)論1)采用OVA混合致敏聯(lián)合束縛制動(dòng)成功建立心理應(yīng)激哮喘大鼠模型,符合支氣管哮喘肝失疏泄、肝肺失和證候?qū)W特征。2)心理應(yīng)激通過(guò)誘導(dǎo)哮喘HPA軸功能失調(diào),GR表達(dá)下調(diào),降低CORT抗炎作用;誘導(dǎo)Th2、Th17免疫偏移,抑制Th1炎癥反應(yīng),引起Th1/Th2免疫失衡、氣道免疫炎癥增強(qiáng),從而促進(jìn)哮喘發(fā)生、發(fā)展。3)調(diào)肝理肺法可顯著改善哮喘心理應(yīng)激哮喘大鼠焦慮、抑郁程度;同時(shí)降低心理應(yīng)激哮喘大鼠血清、支氣管肺泡灌洗液HPA軸相關(guān)指標(biāo),增加肺組織GR表達(dá),調(diào)節(jié)Th細(xì)胞平衡,降低氣道炎癥;提示調(diào)肝理肺法可能通過(guò)調(diào)控HPA軸功能,減輕氣道炎癥反應(yīng),從而實(shí)現(xiàn)對(duì)心理應(yīng)激哮喘的治療作用。
[Abstract]:Research objective asthma is a typical psychosomatic disease. Psychological stress plays an important role in the process of disease. Previous studies have shown that emotion can cause neuroendocrine immune imbalance, and the HPA axis is one of its main regulating pathways. The expression of corticosteroid receptor and the downregulation of function and the decrease of hormone sensitivity. This study simulated the pathogenic characteristics of clinical emotion, established the model of psychological stress asthmatic rats, and detected the changes of the HPA axis function and the correlation with the airway inflammation reaction in the psychological stress asthmatic rats. The rats of stress asthma were intervened to observe the changes of macro characterization, behavior and serum, serum, lavage fluid and lung tissue related indexes in the model rats, and explore the regulating effect of regulating liver and lung method on the HPA axis and airway inflammation in the rats with psychological stress asthma. On the base of this, male SD rats were randomly divided into normal group, stress group, asthma group, stress asthma group, western medicine group and traditional Chinese medicine group. The characteristics of clinical emotion were simulated and the method of combination of ovalbumin sensitization combined with restraint braking was used to establish the model of psychological stress asthma rats. The model was modeled by the objective indexes of macroscopic characterization, behavior and lung ventilation. By detecting serum, CRH, ACTH, CORT levels, GR expression in lung tissue and the content of airway inflammatory factors in the lung tissue, the changes in the function of HPA axis and the correlation with the airway inflammation were studied in the model rats, and the effects of asthma Ning Granule on the psychological stress asthma rats were intervened, and the serum and BALF were observed in comparison with the other groups. The changes in the correlation index concentration, expression of positive cells and protein expression in the lung tissue, study the regulating effect of liver regulating method on the HPA axis function and the imbalance of immune inflammation in the rats with psychological stress asthma. Results 1) the model of psychological stress asthma was established by OVA mixed sensitization combined with binding brake. The results showed that the stress asthma was stressed. Group rats were characterized by diminished interest, dark hair, loose stool, loose stool, reduced diet, and slow weight gain. In the process of stimulation, there were scratching and snout, abdominal skin, choking, sneezing and laryngeal wheezing. Compared with the normal group, the level of stress in the stress asthmatic group was significantly decreased (P0.05, P0.01); the preference of sucrose water, The time of the suspension experimental struggle decreased (P0.01); the duration of the suspension experiment was significantly increased (P0.01); pulmonary ventilation function PEF, FEF25-75%, FEV0.3/FVC (P0.05, P0.01, P0.01), airway resistance Ri, Re significantly increased (P0.05, P0.01); venous blood eosinophils increased; lung tissue inflammatory cells infiltrated obviously. Compared with the stress group or asthma group, The symptoms, signs and related indexes of the stress asthma group were significantly worse in.2). Compared with the normal group, the serum levels of CRH, ACTH, CORT and IgE in the stress asthmatic group were significantly higher (P0.01), and the IFN- gamma content was significantly decreased (P0.01). The expression and content of GR positive in lung tissue decreased. The expression of IFN- gamma positive cells was significantly decreased (P0.01), and the expression of IFN- gamma in the lung tissue decreased slightly, and the difference was not statistically significant (P0.05). Compared with the stress group or asthma group, the HPA axis and airway inflammation related indexes in the stress asthma group were different. The correlation study showed that CORT and IL-4, IL-17, IgE were positive. The correlation was negatively correlated with IFN-y, while the level of GR in lung tissue was opposite to that of IFN-y, but negatively correlated with IL-4 and IL-17. The correlation between CORT content and behavioral indexes of psychological stress asthma rats showed that the content of CORT was negatively correlated with sugar water consumption and sugar water, while the content of GR and the consumption of sugar and water in lung tissue were related. There was a positive correlation between the proportion of sugar and water, the number of turning times and horizontal vertical movement; but it had a negative correlation with the rest time, and the correlation was weak.3). After the treatment of asthma Ning Granule, the early activity of the rats in the Chinese medicine group decreased, the reaction was slow, the hair was dark, the later mood improved, the diet increased, the interest in the surroundings increased, and the frequency of respiration increased during the stimulation process. Quickly, the contraction of the abdominal muscles deepened and accelerated. Compared with the model group, the horizontal movement of the rats in the traditional Chinese medicine group increased significantly (P0.01); the preference degree of the sucrose water, the struggle times of the suspension experiment increased obviously (P0.01, P0.01); the time of the suspension experiment was significantly reduced (P0.01); the lung ventilation function group of Chinese medicine group rats PEF, FEF25-75% and FEV0.3/FVC were slightly higher, while Ri, Re decreased, the difference was statistically significant (P0.01), the eosinophil of venous blood was significantly decreased (P0.01), and the infiltration of inflammatory cells in the lung tissue decreased significantly. The serum of the rats in the Chinese medicine group, CRH, ACTH, CORT in BALF were significantly lower than those in the stress asthma group (P0.01), and the positive expression of lung tissue increased significantly ( P0.01), with a slight increase in content, there was no statistical difference (P0.05), but serum, BALF and bronchopulmonary IL-4, IL-17 content and positive cell expression were significantly decreased (P0.01), and the expression of IFN- gamma positive cells in lung tissue decreased significantly (P0.01), but the content was slightly lower, the difference was not statistically significant (P0.05). The correlation study showed the CORT content. There was a negative correlation with IFN- gamma, but positive correlation with IgE, IL-4, IL-17 and other inflammatory markers, while the GR content in lung tissue was positively correlated with the level of IFN- gamma, but negatively correlated with the inflammatory markers such as IL-4 and IL-17, while the correlation between CORT content and rat behavior found that the CORT content was negatively correlated with sugar water consumption, sugar water ratio, the number of flipping times and the number of vertical movements. There is a positive correlation with the rest time, but the correlation is weak, but the correlation is weak, no statistical difference (P0.05). Conclusion 1) the use of OVA hybrid sensitization combined with binding brake to establish the model of psychological stress asthma rats, in line with bronchial asthma liver loss, liver and lung syndrome.2) psychological stress through the induction of asthma HPA axis dysfunction, GR table Down regulation, reducing the anti-inflammatory effect of CORT, inducing Th2, Th17 immunization, inhibiting Th1 inflammatory reaction, causing Th1/Th2 immune imbalance, enhancing airway immunity and inflammation, promoting the occurrence of asthma and developing.3), regulating liver and lung therapy can significantly improve the anxiety and depression degree of asthmatic asthmatic rats; meanwhile, reduce the serum and bronchi of psychological stress asthmatic rats. The HPA axis related index of alveolar lavage fluid increases the expression of GR in lung tissue, regulates the balance of Th cells and reduces airway inflammation, suggesting that the liver regulating method may reduce airway inflammation by regulating the function of HPA axis, thus achieving the therapeutic effect on psychological stress asthma.
【學(xué)位授予單位】：北京中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R256.12;R-332

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