本文選題：哮喘 + 中性粒細(xì)胞　； 參考：《廣西醫(yī)科大學(xué)》2014年博士論文

【摘要】：支氣管哮喘（哮喘）是由多種炎癥細(xì)胞（嗜酸性粒細(xì)胞、肥大細(xì)胞、T淋巴細(xì)胞、中性粒細(xì)胞等）、氣道結(jié)構(gòu)細(xì)胞（氣道上皮細(xì)胞、平滑肌細(xì)胞、成纖維細(xì)胞）以及多種細(xì)胞因子共同參與的氣道慢性炎癥性疾病。哮喘發(fā)病機(jī)制復(fù)雜，至今尚未完全闡明。氣道慢性炎癥是哮喘的本質(zhì)，研究哮喘氣道炎癥發(fā)生、發(fā)展機(jī)制對(duì)于哮喘防治具有重要臨床指導(dǎo)意義。隨著對(duì)哮喘氣道炎癥的深入研究，人們對(duì)哮喘的認(rèn)識(shí)已不再局限于嗜酸性粒細(xì)胞氣道炎癥，而更強(qiáng)調(diào)哮喘是多種細(xì)胞共同參與的氣道慢性炎癥。目前研究顯示哮喘的氣道炎癥存在不同的亞型，可分為嗜酸細(xì)胞性哮喘（eosinophilicasthma，EA）和非嗜酸細(xì)胞性哮喘（non-eosinophilic asthma，NEA），其中半數(shù)以上的非嗜酸細(xì)胞性哮喘為中性粒細(xì)胞性哮喘（neutrophilic asthma，NA）。但哮喘氣道中性粒細(xì)胞增多的機(jī)制目前尚未完全闡明。輔助性T淋巴細(xì)胞(T helper cell，Th細(xì)胞)在哮喘氣道慢性炎癥中發(fā)揮著重要的免疫調(diào)節(jié)作用。Th2細(xì)胞占優(yōu)勢(shì)的Th1/Th2細(xì)胞失衡被認(rèn)為是變應(yīng)性哮喘氣道嗜酸細(xì)胞性炎癥形成及發(fā)展的免疫學(xué)基礎(chǔ)。但有研究發(fā)現(xiàn)非變應(yīng)性哮喘患者存在不依賴(lài)于Th2細(xì)胞免疫學(xué)機(jī)制介導(dǎo)的氣道中性粒細(xì)胞炎癥。近年來(lái)發(fā)現(xiàn)Th17細(xì)胞及其效應(yīng)因子IL-17在支氣管哮喘等變態(tài)反應(yīng)性疾病中發(fā)揮重要作用， IL-17具有強(qiáng)大的募集、活化中性粒細(xì)胞的作用，可能是導(dǎo)致氣道中性粒細(xì)胞增多的機(jī)制之一。但Th17細(xì)胞在不同氣道炎癥類(lèi)型哮喘發(fā)病中的作用是否相同，在中性粒細(xì)胞性哮喘發(fā)病機(jī)制中是否存在Th17細(xì)胞分化優(yōu)勢(shì)，鮮有報(bào)道。我們前期動(dòng)物實(shí)驗(yàn)通過(guò)建立中性粒細(xì)胞哮喘小鼠模型進(jìn)行研究，，結(jié)果顯示中性粒細(xì)胞哮喘小鼠存在強(qiáng)烈的Th17細(xì)胞免疫、中度Th2細(xì)胞免疫反應(yīng)，Th17細(xì)胞通過(guò)IL-17介導(dǎo)哮喘小鼠中性粒細(xì)胞性氣道炎癥的發(fā)生，并通過(guò)上調(diào)轉(zhuǎn)錄因子RORγt促進(jìn)Th17細(xì)胞分化，中性粒細(xì)胞哮喘小鼠體內(nèi)已分化的Th17細(xì)胞在機(jī)體高IL-7環(huán)境下，依賴(lài)JAK/STAT5信號(hào)途徑激活維持其存活狀態(tài)。但在人類(lèi)中性粒細(xì)胞性哮喘發(fā)病機(jī)制中是否存在同樣的Th17細(xì)胞分化優(yōu)勢(shì)是本課題所要研究的第一個(gè)問(wèn)題。 糖皮質(zhì)激素作為治療哮喘最有效的抗炎藥物主要通過(guò)減少T淋巴細(xì)胞活化及抑制細(xì)胞因子的產(chǎn)生減輕氣道炎癥反應(yīng)。對(duì)于多數(shù)哮喘患者以吸入糖皮質(zhì)激素（ICS）為基礎(chǔ)的規(guī)范化哮喘治療可達(dá)到哮喘控制，但仍有部分患者并沒(méi)有取得預(yù)期的效果，少數(shù)患者癥狀持續(xù)存在、甚至惡化，隨著對(duì)哮喘氣道炎癥的深入研究，發(fā)現(xiàn)激素治療效果欠佳的哮喘患者多為中性粒細(xì)胞哮喘。近年有研究發(fā)現(xiàn)，糖皮質(zhì)激素不能抑制Th17細(xì)胞及其效應(yīng)因子IL-17介導(dǎo)的氣道中性粒細(xì)胞炎癥，但亦有研究顯示糖皮質(zhì)激素治療可下調(diào)IL-17表達(dá)，我們前期動(dòng)物實(shí)驗(yàn)研究亦發(fā)現(xiàn)地塞米松可通過(guò)下調(diào)中性粒細(xì)胞哮喘小鼠Th17細(xì)胞特異性轉(zhuǎn)錄因子RORγt表達(dá)水平，降低Th17細(xì)胞數(shù)量及其相關(guān)細(xì)胞因子IL-17水平，部分減輕哮喘小鼠氣道中性粒細(xì)胞炎癥。但地塞米松對(duì)Th17細(xì)胞存活延長(zhǎng)的狀態(tài)及氣道中性粒細(xì)胞凋亡率無(wú)明顯影響。目前，對(duì)于糖皮質(zhì)激素在Th17細(xì)胞介導(dǎo)的哮喘氣道炎癥中的作用存在不同的研究結(jié)果。我們將在探討Th17細(xì)胞與兒童中性粒細(xì)胞性哮喘氣道炎癥相關(guān)性的基礎(chǔ)之上，進(jìn)一步探討糖皮質(zhì)激素對(duì)Th17細(xì)胞介導(dǎo)的兒童中性粒細(xì)胞性哮喘氣道炎癥的作用。本研究將初步闡明Th17細(xì)胞在兒童中性粒細(xì)胞性哮喘發(fā)病中的作用，為兒童中性粒細(xì)胞性哮喘的治療尋求新線索及提供理論依據(jù)，有助于哮喘的個(gè)體化治療。研究分為兩部分進(jìn)行： 第一部分Th17細(xì)胞及相關(guān)細(xì)胞因子與兒童中性粒細(xì)胞性哮喘氣道炎癥相關(guān)性分析 目的探討Th17細(xì)胞及相關(guān)細(xì)胞因子與兒童中性粒細(xì)胞性哮喘氣道炎癥的相關(guān)性。 方法隨機(jī)選取28例未應(yīng)用糖皮質(zhì)激素治療急性發(fā)作期哮喘患兒，進(jìn)行肺功能測(cè)定、誘導(dǎo)痰細(xì)胞分類(lèi)計(jì)數(shù)，并根據(jù)誘導(dǎo)痰液細(xì)胞學(xué)分類(lèi)分成三組：嗜酸細(xì)胞性哮喘（EA組）12例；中性粒細(xì)胞哮喘（NA組）10例；非嗜酸細(xì)胞非中性粒細(xì)胞性哮喘（NEA+NNA組）共6例，并選取健康對(duì)照組(HC組)10例，采用流式細(xì)胞術(shù)檢測(cè)外周血Th17、Th2細(xì)胞及Th17細(xì)胞上Ki-67、STAT5、BCL-2的表達(dá)；實(shí)時(shí)熒光定量聚合酶鏈反應(yīng)（Real-time PCR）檢測(cè)外周血單個(gè)核細(xì)胞RORγt-mRNA的表達(dá)；ELISA方法檢測(cè)痰上清液、血漿、經(jīng)PMA刺激的PBMC培養(yǎng)上清液IL-17濃度、痰上清液IL-8、IL-5濃度，并進(jìn)行相關(guān)性分析。 結(jié)果1、NA組外周血Th細(xì)胞中Th17細(xì)胞百分比較EA組、NEA+NNA組、HC組均明顯增高(p0.01)，EA組、NEA+NNA組Th17細(xì)胞百分比較HC組增高(p0.01)，EA組與NEA+NNA組間Th17細(xì)胞百分比比較差異無(wú)統(tǒng)計(jì)學(xué)意義(p0.05)；EA組外周血Th細(xì)胞中Th2細(xì)胞百分比較NA組、NEA+NNA組、HC組均增高(p0.01)，NA組、NEA+NNA組Th2細(xì)胞百分比較HC組增高(p 0.01)，NA組與NEA+NNA組間Th2細(xì)胞百分比比較差異無(wú)統(tǒng)計(jì)學(xué)意義(p0.05)；2、NA組外周血Th17細(xì)胞Ki-67表達(dá)陽(yáng)性率較EA組、NEA+NNA組、HC組均明顯增高(p0.01)，EA組、NEA+NNA組Th17細(xì)胞Ki-67表達(dá)陽(yáng)性率較HC組增高(p0.01)，EA組與NEA+NNA組間Th17細(xì)胞Ki-67表達(dá)陽(yáng)性率比較差異無(wú)統(tǒng)計(jì)學(xué)意義(p0.05)；NA組外周血Th17細(xì)胞STAT5表達(dá)陽(yáng)性率較EA組、NEA+NNA組、HC組均明顯增高(p0.01)，EA組、NEA+NNA組Th17細(xì)胞STAT5表達(dá)陽(yáng)性率較HC組增高(p0.01)，EA組與NEA+NNA組間Th17細(xì)胞STAT5表達(dá)陽(yáng)性率比較差異無(wú)統(tǒng)計(jì)學(xué)意義(p0.05)；NA組外周血Th17細(xì)胞BCL-2表達(dá)陽(yáng)性率較EA組、NEA+NNA組、HC組均明顯增高(p0.01)，EA組、NEA+NNA組與HC組間Th17細(xì)胞BCL-2表達(dá)陽(yáng)性率比較差異無(wú)統(tǒng)計(jì)學(xué)意義(p0.05)；3、NA組外周血單個(gè)核細(xì)胞RORγt-mRNA表達(dá)較EA組、NEA+NNA組、HC組均明顯增高(p0.01)，EA組、NEA+NNA組RORγt-mRNA表達(dá)較HC組增高(p0.01)，EA組與NEA+NNA組間RORγt-mRNA表達(dá)比較差異無(wú)統(tǒng)計(jì)學(xué)意義(p0.05)；4、NA組痰上清液IL-17濃度較EA組、NEA+NNA組、HC組均明顯增高(p0.01)，EA組、NEA+NNA組痰上清液IL-17濃度較HC組增高(p 0.01)，EA組與NEA+NNA組間痰上清液IL-17濃度比較差異無(wú)統(tǒng)計(jì)學(xué)意義(p0.05)；EA組痰上清液IL-5濃度較NA組、NEA+NNA組、HC組均增高(p 0.01)，NA組、NEA+NNA組痰上清液IL-5濃度較HC組增高(p 0.01)，NA組與NEA+NNA組間痰上清液IL-5濃度比較差異無(wú)統(tǒng)計(jì)學(xué)意義(p0.05)；NA組痰上清液IL-8濃度較EA組、NEA+NNA組、HC組均明顯增高(p 0.01)，EA組、NEA+NNA組痰上清液IL-8濃度較HC組增高(p 0.01)，EA組與NEA+NNA組間痰上清液IL-8濃度比較差異無(wú)統(tǒng)計(jì)學(xué)意義(p0.05)；5、NA組、EA組、NEA+NNA組與HC組間血漿IL-17濃度比較差異無(wú)統(tǒng)計(jì)學(xué)意義(p0.05)，但經(jīng)PMA刺激培養(yǎng)后，NA組PBMC培養(yǎng)上清液IL-17濃度較EA組、NEA+NNA組、HC組均明顯增高(p0.01)，EA組、NEA+NNA組PBMC培養(yǎng)上清液IL-17濃度較HC組增高(p0.01)，EA組與NEA+NNA組間PBMC培養(yǎng)上清液IL-17濃度比較差異無(wú)統(tǒng)計(jì)學(xué)意義(p0.05)；同時(shí)，經(jīng)PMA刺激培養(yǎng)后，NA組、EA組、NEA+NNA組PBMC培養(yǎng)上清液IL-17濃度較血漿IL-17濃度均明顯增高(p均0.01)，HC組PBMC培養(yǎng)上清液IL-17濃度與血漿IL-17濃度比較差異無(wú)統(tǒng)計(jì)學(xué)意義(p0.05)。6、哮喘患兒誘導(dǎo)痰NEU百分比與外周血Th細(xì)胞中Th17細(xì)胞比例及痰上清IL-17、IL-8、PBMC培養(yǎng)上清液IL-17濃度呈明顯正相關(guān)；哮喘患兒誘導(dǎo)痰NEU百分比與FEV1%pred、PEF%pred呈明顯負(fù)相關(guān)；哮喘患兒誘導(dǎo)痰EOS百分比與外周血Th細(xì)胞中Th2細(xì)胞比例及痰上清IL-5濃度呈明顯正相關(guān)。 結(jié)論1、Th17細(xì)胞和Th2細(xì)胞均不同程度共同參與兒童哮喘的發(fā)病。兒童中性粒細(xì)胞性哮喘外周血Th17細(xì)胞、特異性轉(zhuǎn)錄因子RORγt表達(dá)及痰液IL-17水平均明顯高于嗜酸細(xì)胞性哮喘及非嗜酸細(xì)胞非中性粒細(xì)胞性哮喘，并且與誘導(dǎo)痰中性粒細(xì)胞比例呈正相關(guān)。提示Th17細(xì)胞可能通過(guò)IL-17介導(dǎo)中性粒細(xì)胞性哮喘氣道炎癥的發(fā)生。2、兒童中性粒細(xì)胞性哮喘體內(nèi)已分化的Th17細(xì)胞存活延長(zhǎng)，可能與JAK/STAT5信號(hào)途徑激活有關(guān)。 第二部分吸入糖皮質(zhì)激素對(duì)Th17細(xì)胞介導(dǎo)的兒童中性粒細(xì)胞性哮喘氣道炎癥的影響 目的探討吸入糖皮質(zhì)激素對(duì)Th17細(xì)胞介導(dǎo)的兒童中性粒細(xì)胞性哮喘氣道炎癥的影響 方法第一部分研究對(duì)象中的中性粒細(xì)胞性哮喘患兒(NA組)10例，在醫(yī)師指導(dǎo)下規(guī)則吸入糖皮質(zhì)激素治療3個(gè)月后，進(jìn)行肺功能測(cè)定、誘導(dǎo)痰細(xì)胞分類(lèi)計(jì)數(shù)；采用流式細(xì)胞術(shù)檢測(cè)外周血Th17細(xì)胞及Th17細(xì)胞上Ki-67、的表達(dá)；實(shí)時(shí)熒光定量聚合酶鏈反應(yīng)（Real-time PCR）檢測(cè)外周血單個(gè)核細(xì)胞RORγt-mRNA的表達(dá)；ELISA方法檢測(cè)痰上清液、PBMC培養(yǎng)上清液IL-17濃度。 結(jié)果1、NA組規(guī)則吸入糖皮質(zhì)激素治療后FEV1%pred、PEF%pred均較治療前明顯增高(均p 0.01)，但仍低于健康對(duì)照組（P均0.05）；2、NA組規(guī)則吸入糖皮質(zhì)激素治療后誘導(dǎo)痰NEU百分比較治療前明顯降低(p0.01)，但仍高于健康對(duì)照組（P0.05）；3、NA組規(guī)則吸入糖皮質(zhì)激素治療后外周血Th17細(xì)胞及Th17細(xì)胞Ki-67表達(dá)均較治療前明顯降低(均p0.01)，但仍高于健康對(duì)照組（均P0.01）；4、NA組規(guī)則吸入糖皮質(zhì)激素治療后外周血單個(gè)核細(xì)胞RORγt-mRNA表達(dá)較治療前明顯降低(p0.01)，與健康對(duì)照組比較差異無(wú)統(tǒng)計(jì)學(xué)意義(p0.05)；5、NA組規(guī)則吸入糖皮質(zhì)激素治療后痰上清液及PBMC培養(yǎng)上清液IL-17濃度均較治療前降低(分別為：p0.05，p 0.01)，但仍高于健康對(duì)照組（均P 0.01）。 結(jié)論吸入糖皮質(zhì)激素可通過(guò)下調(diào)特異性轉(zhuǎn)錄因子RORγt的表達(dá)水平，在一定程度上降低外周血Th17細(xì)胞及痰液IL-17水平，減輕Th17細(xì)胞介導(dǎo)的兒童中性粒細(xì)胞性哮喘氣道炎癥。
[Abstract]:Bronchial asthma (asthma) is a chronic inflammatory disease in the airway, which is involved in a variety of inflammatory cells (eosinophils, mast cells, T lymphocytes, neutrophils, etc.), airway structural cells (airway epithelial cells, smooth muscle cells, fibroblasts) and a variety of cytokines. The pathogenesis of asthma is complex and has not yet been fully explained. Chronic airway inflammation is the essence of asthma. It is important to study the pathogenesis of airway inflammation in asthma. The development mechanism has important clinical guiding significance for the prevention and treatment of asthma. With the in-depth study of airway inflammation in asthma, people's understanding of asthma is no longer limited to eosinophil airway inflammation, but more emphasis is placed on the common participation of multiple cells in asthma. Chronic airway inflammation. The present study shows that there are different subtypes of airway inflammation in asthma, which can be divided into eosinophilicasthma (EA) and non-eosinophilic asthma (NEA), and more than half of the non eosinophilic asthma is neutrophils (NA), but asthma (neutrophilic asthma, NA). The mechanism of polymorphonuclear neutrophils has not yet been fully elucidated. Auxiliary T lymphocytes (T helper cell, Th cells) play an important immunomodulatory role in the chronic airway inflammation of asthma. The dominant Th1/Th2 cell imbalance in.Th2 cells is considered to be the immunological basis for the formation and development of airway eosinophilic inflammation in allergic asthma. However, some studies have found that patients with non allergic asthma are not dependent on Th2 cell immunological mechanism mediated airway neutrophil inflammation. In recent years, Th17 cells and their effector IL-17 have been found to play an important role in allergic diseases such as bronchial asthma. IL-17 has a strong recruitment, the role of activating neutrophils, and the possibility of activating neutrophils. It is one of the mechanisms leading to the increase of neutrophils in the airway. However, the role of Th17 cells in the pathogenesis of different airway inflammation is the same. It is rarely reported whether there is a differentiation advantage of Th17 cells in the pathogenesis of neutrophils. The results showed that there were strong Th17 cell immunity, moderate Th2 cell immune response, Th17 cells mediated the pathogenesis of neutrophil airway inflammation in asthmatic mice by IL-17, and the differentiation of Th17 cells by up regulation of the transcription factor ROR gamma T, and the differentiated Th17 cells in neutrophil asthmatic mice were in the machine. It is the first issue to study whether there is the same Th17 cell differentiation advantage in the pathogenesis of human neutrophil asthma under the high IL-7 environment, which depends on the activation of the JAK/STAT5 signal pathway.
Glucocorticoid as the most effective anti inflammatory drug for asthma is mainly to reduce airway inflammation by reducing the activation of T lymphocytes and inhibiting the production of cytokines. Standardized asthma therapy based on inhaled corticosteroid (ICS) for most asthmatic patients can achieve asthma control, but some patients still have no expectations. In recent years, some studies have found that glucocorticoids do not inhibit Th17 cells and their effects on airway neutrophil inflammation mediated by IL-17, but there are also a few studies. The study showed that glucocorticoid therapy could reduce the expression of IL-17. We also found that dexamethasone could reduce the expression of ROR gamma t in the Th17 cell specific transcription factor of neutrophils, reduce the number of Th17 cells and the related cytokines IL-17 level, and partly reduce the airway neutrality of asthmatic mice. But dexamethasone has no significant effect on the prolonged survival of Th17 cells and the rate of apoptosis in airway neutrophils. At present, there are different results on the role of glucocorticoid in the airway inflammation mediated by Th17 cells. We will explore the correlation between Th17 cells and airway inflammation in children with neutrophils. On the basis of this, the effect of glucocorticoid on airway inflammation in children with neutrophils induced by Th17 cells is further explored. This study will preliminarily clarify the role of Th17 cells in the pathogenesis of neutrophil asthma in children and provide a new clue and theoretical basis for the treatment of neutrophilic asthma in children and help to asthma. The individualized treatment of asthma is divided into two parts:
Part 1 Correlation Analysis of Th17 cells and related cytokines with airway inflammation in children with neutrophilic asthma
Objective to explore the correlation between Th17 cells and related cytokines and airway inflammation in children with neutrophilic asthma.
Methods 28 cases of acute exacerbation of asthma were randomly selected, and the pulmonary function was measured and the sputum cell classification was induced. According to the induced sputum cytology, 12 cases were divided into three groups: 12 cases of eosinophilic asthma (group EA), 10 cases of neutrophil asthma (group NA) and non eosinophilic non neutrophil asthma (NE In group A+NNA) 6 cases were selected and 10 cases of healthy control group (group HC) were selected. Flow cytometry was used to detect the expression of Ki-67, STAT5, BCL-2 on the peripheral blood Th17, Th2 and Th17 cells, and the real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the expression of ROR gamma t-mRNA in peripheral blood mononuclear cells. PBMC stimulated IL-17 concentration in supernatant, IL-8 and IL-5 concentration in sputum supernatant and correlation analysis.
Results 1, the percentage of Th17 cells in peripheral blood Th cells in group NA was significantly higher than that in group EA, NEA+NNA group and HC group (P0.01), EA group and NEA+NNA group Th17 cell percentage was higher than HC group (P0.01). Group HC was increased (P0.01), group NA and group NEA+NNA were higher than group HC (P 0.01), and there was no significant difference in the percentage of Th2 cells between NA and NEA+NNA groups (P0.05). 2. Compared with group HC (P0.01), there was no significant difference in the positive rate of Ki-67 expression in Th17 cells between group EA and NEA+NNA group (P0.05), and the positive rate of STAT5 expression of Th17 cell STAT5 in peripheral blood of NA group was higher than that in EA group. The positive rate of STAT5 expression was not statistically significant (P0.05), and the positive rate of BCL-2 expression in Th17 cells in group NA was significantly higher than that in group EA, NEA+NNA group and HC group (P0.01), and there was no significant difference between the EA group, NEA+NNA group and HC group. The expression of ROR y in group NEA+NNA and group HC increased significantly (P0.01), and the expression of ROR gamma t-mRNA in group EA and NEA+NNA was higher than that in group HC (P0.01), and there was no statistical difference between EA group and NEA+NNA group. 4. There was no significant difference in the concentration of IL-17 in the sputum supernatant between the EA group and the NEA+NNA group (P0.05). The concentration of IL-5 in the sputum supernatant in the EA group was higher than that in the NA group, the NEA+NNA group and the HC group increased (P 0.01). The concentration of the sputum supernatant was higher than that of the group HC (0.01). The concentration of the sputum supernatant was higher than that of the group (0.01). There was no statistical significance (P0.05), and the concentration of IL-8 in the sputum supernatant in group NA was higher than that in group EA, NEA+NNA and HC (P 0.01), EA and NEA+NNA group was higher than that of HC group in group NEA+NNA (P 0.01). 5 There was no statistical significance (P0.05), but after PMA stimulation, the concentration of IL-17 in PBMC culture supernatant of NA group was higher than that of EA group, NEA+NNA group and HC group increased significantly (P0.01), EA group and NEA+NNA group were higher than those in the group of PBMC culture supernatant. P0.05); at the same time, after PMA stimulation, the concentration of IL-17 in the PBMC culture supernatant of group NA, EA and NEA+NNA was significantly higher than that of plasma IL-17 (P 0.01). There was no significant difference between the IL-17 concentration of the PBMC culture supernatant and the plasma concentration in HC group. The proportion and the IL-17 concentration of the supernatant IL-17, IL-8, PBMC culture supernatant was positively correlated, the percentage of NEU in the induced sputum in children with asthma was negatively correlated with FEV1%pred and PEF%pred, and the percentage of EOS in the induced sputum in children with asthma was positively correlated with the proportion of Th2 cells in the peripheral Th cells and the IL-5 concentration of the sputum.
Conclusion 1, Th17 cells and Th2 cells are all involved in the pathogenesis of childhood asthma. The expression of Th17 cells in peripheral blood, the specific transcription factor ROR gamma T and the level of IL-17 in sputum in children with neutrophils are significantly higher than those of eosinophilic and non eosinophilic non neutrophilic granulocytic asthma, and it is also associated with the induction of sputum neutrophils. The proportion is positive correlation. It suggests that Th17 cells may mediate the occurrence of airway inflammation in neutrophil asthma by IL-17, and the survival of the differentiated Th17 cells in neutrophilic asthma in children may be prolonged, which may be related to the activation of the JAK/STAT5 signal pathway.
The second part is the effect of inhaled corticosteroids on airway inflammation in Th17 cell mediated neutrophilic asthma in children.
Objective to investigate the effects of inhaled corticosteroids on airway inflammation induced by Th17 cells in children with neutrophilic asthma.
Methods in the first part of the study, 10 children with neutrophils (NA) were studied. After 3 months of regular inhalation of glucocorticoid under the guidance of the physician, the lung function was measured and the phlegm cell classification was induced. Flow cytometry was used to detect the expression of Ki-67 on the peripheral blood and Th17 cells; real-time fluorescence quantitative polymerization was used. Real-time PCR was used to detect the expression of ROR t-mRNA in peripheral blood mononuclear cells. ELISA method was used to detect sputum supernatant and PBMC culture supernatant IL-17 concentration.
Results 1, FEV1%pred and PEF%pred were significantly higher in group NA after inhaling corticosteroid than before treatment (all P 0.01), but still lower than that in healthy control group (P 0.05); 2, group NA was significantly lower before treatment (P0.01) after inhaled corticosteroid therapy, but still higher than that in healthy control group (P0.05); 3, NA group rule inhalation. The expression of Ki-67 in peripheral blood Th17 cells and Th17 cells in peripheral blood was significantly lower than before treatment (all P0.01), but still higher than that in the healthy control group (all P0.01). 4, the expression of ROR y t-mRNA in peripheral blood mononuclear cells of group NA was significantly lower than that before treatment (P0.01), and there was no difference compared with that of the healthy control group. Study significance (P0.05); 5, the concentration of IL-17 in the sputum supernatant and PBMC culture supernatant after inhaled corticosteroid treatment in group NA was lower than that before treatment (P0.05, P 0.01), but still higher than that in the healthy control group (P 0.01).
Conclusion inhaled corticosteroids can reduce the level of expression of specific transcription factor ROR gamma T, to a certain extent, reduce the level of IL-17 in peripheral blood Th17 cells and sputum, and reduce airway inflammation in children with neutrophilic granulocytic asthma mediated by Th17 cells.

【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R725.6

【參考文獻(xiàn)】

相關(guān)期刊論文 前5條

1 張毓嫻;農(nóng)光民;廖競(jìng);蔣敏;梁秀安;劉靜;;誘導(dǎo)痰液細(xì)胞學(xué)分析在兒童支氣管哮喘臨床分型中的應(yīng)用[J];實(shí)用兒科臨床雜志;2010年02期

2 全國(guó)兒科哮喘協(xié)作組;2000年與1990年兒童支氣管哮喘患病率的調(diào)查比較[J];中華結(jié)核和呼吸雜志;2004年02期

3 趙瑩,孔靈菲;糖皮質(zhì)激素對(duì)支氣管哮喘患者痰嗜酸粒細(xì)胞和中性粒細(xì)胞凋亡的影響[J];中華結(jié)核和呼吸雜志;2004年04期

4 周慶濤,孫永昌,姚婉貞;重度支氣管哮喘患者氣道炎癥及其與白細(xì)胞介素17的關(guān)系[J];中華結(jié)核和呼吸雜志;2005年09期

5 周辰;陳軍浩;殷凱生;;糖皮質(zhì)激素對(duì)支氣管哮喘小鼠Th 17細(xì)胞的調(diào)節(jié)作用[J];中國(guó)現(xiàn)代醫(yī)學(xué)雜志;2010年22期

本文編號(hào)：1860696