本文選題：吸入性糖皮質(zhì)激素 + 哮喘�。� 參考：《南華大學(xué)》2014年碩士論文

【摘要】：目的 通過對亞急性的哮喘大鼠模型進(jìn)行的實驗研究，了解吸入性糖皮質(zhì)激素藥物使用后，大鼠的氣道炎癥及氣道阻力發(fā)生的變化，以及大鼠肺組織的甲殼質(zhì)酶3樣蛋白質(zhì)1(Chitinase-3-like-1protein，YKL-40)、白細(xì)胞介素-8（Interleukin-8，IL-8）表達(dá)變化，了解吸入性糖皮質(zhì)激素對哮喘大鼠肺組織中YKL-40mRNA及蛋白、IL-8mRNA表達(dá)水平的影響。 方法 1.40只清潔健康，約1月齡，體重150-170克的雄性SD大鼠，隨機(jī)分配為以下四個組別：正常對照組（Control組）、哮喘模型組（Model組）、地塞米松組（DXM組），布地奈德組（BUD組），每一組分別劃分10只SD大鼠。 2. Model組大鼠、DXM組大鼠、BUD組大鼠在實驗進(jìn)展的第1天、第8天給予腹腔注射10㳠的卵清蛋白(OVA)溶液1毫升，稀釋有免疫佐劑氫氧化鋁0.1克。在實驗第15天，用2㳠的卵清蛋白溶液4毫升對大鼠進(jìn)行霧化吸入以激發(fā)哮喘，每1次激發(fā)的時間為10分鐘，1次/天，連續(xù)共兩周。其中DXM組大鼠在先給予腹腔注射地塞米松注射液（劑量按0.5毫克/千克計算）后半小時再予以霧化吸入激發(fā)OVA，,而布地奈德組先給予布地奈德來泵霧化吸入(按1mg/2ml使用)。相對應(yīng)正常對照組則采用生理鹽水代替卵清蛋白溶液進(jìn)行腹腔注射與霧化吸入以致敏與激發(fā)哮喘產(chǎn)生。 3.在實驗第28天，四組大鼠先予以檢測氣道阻力值，在檢測結(jié)束后,采用剪斷大鼠腹主動脈的方法來處死動物，在獲取左右兩側(cè)肺組織后，取左肺組織用實時熒光定量PCR方法（qPCR）來檢測肺組織中YKL-40與IL-8mRNA的轉(zhuǎn)錄水平，并通過WesternBlot檢測方法來檢測大鼠肺組織中YKL-40的蛋白表達(dá)水平；取大鼠右側(cè)肺組織通過HE染色病理檢查來分析四組大鼠中氣道炎癥的變化情況。 結(jié)果 1.成功建立亞急性哮喘大鼠模型：實驗第14天開始予以2㳠卵清蛋白溶液霧化吸入后，隨著次數(shù)增加逐漸出現(xiàn)以下哮喘發(fā)作癥狀：面部搔抓、呼吸頻率加快、點頭呼吸、面部紫紺，大小便失禁等，于實驗第28天予檢測其氣道阻力變化，氣道阻力結(jié)果提示哮喘模型組的氣道阻力均有顯著增加，與正常對照組比較有顯著的差異（P＜0.05）；通過大鼠肺組織的HE染色病理切片進(jìn)行觀察：Model組肺組織中支氣管與肺泡周圍嗜酸性粒細(xì)胞增多，氣道內(nèi)有較多粘性分泌物，支氣管管壁稍顯增厚。 2.BUD組大鼠肺組織氣道炎癥的變化:與哮喘大鼠Model組比較，BUD組炎癥細(xì)胞及粘性分泌物均明顯減少。DXM組同BUD組對比，形態(tài)學(xué)差異不大。control組中未發(fā)現(xiàn)明顯炎癥反應(yīng)。 3.BUD組大鼠氣道阻力變化:經(jīng)過鹽酸組胺霧化吸入后，哮喘大鼠Model組的氣道阻力與Control組比較顯著增高，差異有統(tǒng)計學(xué)意義(P＜0.05)。布地奈德組大鼠氣道阻力和哮喘模型大鼠相比較有顯著降低，有顯著的差異(P＜0.05)，，與正常對照組相比較氣道阻力有明顯增高，有顯著的差異(P＜0.05)，同地塞米松組相比較，發(fā)現(xiàn)沒有顯著差異。 4. BUD大鼠肺組織中YKL-40與IL-8的mRNA變化：哮喘大鼠Model組的肺組織YKL-40mRNA與IL-8的mRNA轉(zhuǎn)錄水平與Control組比較顯著增高(P＜0.05)。BUD組肺組織的YKL-40mRNA與IL-8的mRNA轉(zhuǎn)錄水平與Model組比較顯著降低(P＜0.05)，仍顯著高于正常對照組(P＜0.05)，同地塞米松組對比沒有顯著差異。 5. BUD組大鼠肺組織YKL-40蛋白表達(dá)量變化情況：哮喘模型組大鼠的肺組織YKL-40的蛋白表達(dá)量同正常對照組相比有顯著的升高(P＜0.05)，布地奈德組大鼠的肺組織YKL-40的蛋白表達(dá)量同哮喘模型組相比較有顯著的降低(P＜0.05)，但仍高于正常對照組，同地塞米松組比較沒有顯著的差異。 結(jié)論 1.以卵清蛋白做為抗原可成功建立哮喘大鼠亞急性模型。哮喘模型組大鼠的肺組織中YKL-40的蛋白表達(dá)量同正常對照組相比有顯著的升高，布地奈德組大鼠的肺組織YKL-40的蛋白表達(dá)量同哮喘模型組相比有顯著的降低，但仍高于正常對照組，同地塞米松組比較沒有顯著的差異。 2.哮喘大鼠肺組織YKL-40mRNA及蛋白與IL-8mRNA表達(dá)水平升高，提示細(xì)胞因子YKL-40與IL-8參與了支氣管哮喘發(fā)病過程。 3.霧化吸入布地奈德能夠減輕氣道炎癥與氣道阻力，有效降低大鼠局部肺組織中YKL-40mRNA與IL-8mRNA的轉(zhuǎn)錄，減少YKL-40蛋白表達(dá)。
[Abstract]:objective
The changes in airway inflammation and airway resistance in rats after inhalation of inhaled glucocorticoids, as well as the changes in the expression of the chitinase 3 like protein 1 (Chitinase-3-like-1protein, YKL-40), and the expression of interleukin -8 (Interleukin-8, IL-8) in rat lung tissue were investigated by an experimental study of the subacute asthmatic rat model. Effects of inhaled corticosteroids on expression of YKL-40mRNA and protein and IL-8mRNA in lung tissue of asthmatic rats.
Method
1.40 healthy, 1 month old, and 150-170 g male SD rats were randomly assigned to the following four groups: normal control group (group Control), asthma model group (group Model), dexamethasone group (group DXM), and budesonide group (group BUD), each group was divided into 10 SD rats.
2. Model rats, group DXM rats, group BUD rats on the first day of the experiment, 1 ml of 10? Ovalbumin solution (1 ml) and 0.1 grams of aluminum hydroxide with an immune adjuvant. On the fifteenth day, 2? Ovalbumin solution was used to stimulate the rats to stimulate asthma, and the time of every 1 excitation was 10 minutes. 1 times per day, for a total of two weeks, group DXM rats were first given intraperitoneal injection of dexamethasone injection (dosage according to 0.5 mg / kg) and then atomized and inhaled to stimulate OVA, while budesonide group was given budesonide first to pump inhalation (according to 1mg/2ml). The protein solution was injected intraperitoneally and nebulized so as to sensitize and stimulate asthma.
3. on the twenty-eighth day of the experiment, the four groups of rats first detected the airway resistance. After the end of the test, the rat abdominal aorta was cut to kill the animals. After obtaining the left and right bilateral lung tissues, the left lung tissue was taken to detect the transcriptional level of YKL-40 and IL-8mRNA in the lung tissue by real time fluorescence quantitative PCR method (qPCR), and through WesternBlot examination. The protein expression level of YKL-40 in the lung tissue of rats was detected and the changes of airway inflammation in the four groups of rats were analyzed by the pathological examination of the right lung tissue by HE staining.
Result
1. a rat model of subacute asthma was successfully established: after fourteenth days of experiment, 2 of the ovalbumin solution was inhaled. With the increase of the number of times, the symptoms of asthma attack gradually appeared following the increasing number of times: the facial scratch, the speed of breathing, the head breathing, the cyanosis of the face, the incontinence, and so on. The airway resistance changes and the airway obstruction were detected on the twenty-eighth day of the experiment. The force results showed that the airway resistance of the asthma model group increased significantly, and there was a significant difference compared with the normal control group (P < 0.05). The HE staining pathological sections of the lung tissue of rats were observed: the increased eosinophils around the bronchi and alveoli, more viscous secretions in the airway and the bronchial tube wall in the lung tissue of the Model group were observed. It's thicker.
The changes of airway inflammation in the lung tissue of group 2.BUD rats: compared with the Model group of the asthmatic rats, the inflammatory cells and the sticky secretions in the group BUD significantly decreased the comparison between the group.DXM and the BUD group, and there was no obvious inflammatory reaction in the group of.Control.
The airway resistance changes in the 3.BUD group: after inhalation of histamine hydrochloride, the airway resistance of the Model group was significantly higher in the group of asthma rats than that in the Control group (P < 0.05). The airway resistance of the budesonide group was significantly lower than that of the asthma model rats. There were significant differences (P < 0.05), with the normal control group. Compared with the dexamethasone group, there was no significant difference in airway resistance between the two groups (P < 0.05).
The mRNA changes of YKL-40 and IL-8 in the lung tissue of 4. BUD rats: the mRNA transcriptional level of YKL-40mRNA and IL-8 in the lung tissue of the group Model of the asthmatic rats was significantly higher than that in the Control group (P < 0.05) the transcriptional level of the YKL-40mRNA in the lung tissue of the.BUD group was significantly lower than that in the control group (0.05), still significantly higher than that in the normal control group (< 0.05). There was no significant difference in the comparison with the dexamethasone group.
The changes in the expression of YKL-40 protein in the lung tissue of 5. BUD rats: the protein expression of YKL-40 in the lung tissue of the asthmatic model group was significantly higher than that in the normal control group (P < 0.05). The protein expression of YKL-40 in the lung tissue of the budesonide group was significantly lower than that in the asthma model group (P < 0.05), but still higher than that in the model group (P < 0.05). There was no significant difference between the control group and the dexamethasone group.
conclusion
1. the subacute model of asthmatic rats was successfully established with ovalbumin as antigen. The protein expression of YKL-40 in the lung tissue of the asthmatic model group was significantly higher than that in the normal control group. The protein expression of YKL-40 in the lung tissue of the budesonide group was significantly lower than that of the asthma model group, but it was still higher than the normal control group. There was no significant difference in the group, compared with the dexamethasone group.
2. the expression level of YKL-40mRNA and protein and IL-8mRNA in lung tissue of asthmatic rats increased, suggesting that cytokines YKL-40 and IL-8 are involved in the pathogenesis of bronchial asthma.
3. inhalation of budesonide can reduce airway inflammation and airway resistance, effectively reduce the transcription of YKL-40mRNA and IL-8mRNA in the local lung tissue of rats and reduce the expression of YKL-40 protein.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R725.6

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 賁素琴;袁佩紅;仇亞莉;倪松石;;YKL-40在哮喘小鼠外周血、肺泡灌洗液、氣道上皮中的表達(dá)及其與氣道嗜酸性粒細(xì)胞炎癥的關(guān)系[J];交通醫(yī)學(xué);2012年06期

2 汪浩;武曉蘭;譚紅霞;范曉云;;IL-8和Eotaxin在哮喘大鼠中的表達(dá)及地塞米松的作用[J];安徽醫(yī)科大學(xué)學(xué)報;2012年03期

3 劉中成;張艷芬;;一種大鼠慢性哮喘模型的建立與評價[J];藥學(xué)學(xué)報;2010年06期

本文編號：1968441