本文選題：支氣管哮喘 + 白細胞介素17��； 參考：《瀘州醫(yī)學院》2013年碩士論文

【摘要】：目的：探討B(tài)細胞轉(zhuǎn)錄激活因子(B cell activating transcription factor, BATF)對卵清蛋白(OVA)致急性哮喘小鼠氣道炎癥的調(diào)控機制及其與維甲酸孤兒核受體γt (RORyt)的關(guān)系。方法：(1)將24只健康雌性BALB/c小鼠(SPF級,體重15-18g,周齡68周),隨機分為3組,0.9%生理鹽水對照組(NS組)、哮喘模型組(AS組)、地塞米松治療組(DEX組),每組8只。AS組、DEX組小鼠分別于第0、7、14天用卵清蛋白抗原液(100 μg OVA和lmg氫氧化鋁混合的生理鹽水混懸液)200 ul腹腔內(nèi)注射致敏,NS組用等體積生理鹽水以同種方式代替抗原液。第21天起,NS組以生理鹽水,AS組、DEX組以2%的OVA超聲霧化吸入進行激發(fā),每天1次,每次40分鐘,連續(xù)5天。DEX組于每次激發(fā)前30分鐘,予以地塞米松1. Omg/kg腹腔注射進行干預(yù),NS組、AS組則予以等體積的生理鹽水腹腔注射,時間同上。各組小鼠末次激發(fā)結(jié)束24h后腹腔注射麻醉,行支氣管肺泡灌洗及留取肺組織標本。(2)分別對支氣管肺泡灌洗液(Bronchoalveolar lavage fluid, BALF)中白細胞(WBC)及嗜酸性粒細胞(EOS)、中性粒細胞(PMN)進行計數(shù)；雙抗體夾心酶聯(lián)免疫吸附法(ELISA)檢測上清液中白細胞介素17 (Interleukin 17, IL-17)的表達；肺組織病理切片,HE染色觀察支氣管肺組織炎癥浸潤程度；實時熒光定量聚合酶鏈式反應(yīng)(real time Polymerase Chain Reaction, RT-PCR)分別檢測肺組織中IL-17、BATF、 RORγt在mRNA水平的表達情況。利用SPSS17.0軟件對各組實驗數(shù)據(jù)進行統(tǒng)計學分析,變量以均數(shù)±標準差(x±s)表示,組間變量用單因素方差分析,兩變量的相關(guān)關(guān)系用直線相關(guān)分析,統(tǒng)計學差異均以p0.05界定。結(jié)果： (1)病理組織切片HE染色結(jié)果顯示：NS組小鼠支氣管管壁平滑肌較薄,管腔光滑、規(guī)則,腔內(nèi)未見分泌物積存,氣道黏膜皺襞低平規(guī)則,肺泡間隔寬且清晰,支氣管血管管周及肺泡間隔內(nèi)無明顯炎癥細胞浸潤；AS組小鼠支氣管管壁平滑肌明顯增厚,管腔內(nèi)大量粘液栓形成及氣道黏膜皺襞肥厚增生不規(guī)則,纖毛排列紊亂,致支氣管管腔極度狹窄,多處氣道上皮斷裂、脫落不完整,支氣管血管管周及肺泡間隔內(nèi)大量炎癥細胞浸潤,肺泡間隔增厚明顯,分界不清,炎癥細胞浸潤多以EOS、淋巴細胞及P惻為主；DEX組小鼠支氣管管壁增厚及管腔狹窄的程度均較AS組減輕,管腔內(nèi)分泌物顯著減少,肺泡間隔清晰可見,支氣管血管管周及肺泡間隔內(nèi)炎癥細胞浸潤情況有所改善。(2)AS組BALF中的白細胞總數(shù)、嗜酸性粒細胞和中性粒細胞數(shù)較NS組明顯增多(PO.01)；與AS組相比較,DEX組白細胞總數(shù)、嗜酸性粒細胞及中性粒細胞數(shù)明顯減少(P0.01),但仍比NS組多(P0.05)。 (3)ELISA結(jié)果顯示：AS組BALF中IL一17蛋白濃度明顯高于NS組、DEX組(P0.05)；與AS組相比,DEX組BALF中IL-17蛋白濃度有所降低,差異具有統(tǒng)計學意義(P0.05)。 (4)RT-PCR結(jié)果顯示：AS組肺組織IL-17、BATF、RORγt mRNA的表達水平明顯高于NS組、DEX組,差異均具有統(tǒng)計學意義(PO.05)；與AS組相比,DEX組肺組織IL-17、BATF、RORγt mRNA的表達水平降低,差異具有統(tǒng)計學意義(P0.05)。(5)相關(guān)關(guān)系分析：小鼠肺組織IL-17 mRNA的表達與BATF、 RORγt的表達及BALF中WBC、EOS、PMN計數(shù)呈正相關(guān)(r=0.934,2=0.776, r=0.833,2=0.852,2=0.916,p均0.05)；小鼠肺組織BATF與ROR γ t mRNA表達及BALF中WBC、EOS、PMN計數(shù)呈明顯正相關(guān)(r=0.860,r=0.884, r=0.890,r=0.951, p均0.05)。結(jié)論： (1)在急性哮喘小鼠支氣管肺組織中存在BATF、IL-17、RORγt表達的上調(diào),且三者的表達呈正相關(guān)性。(2) BATF的表達增高可能是引起哮喘小鼠氣道炎癥的關(guān)鍵因素,為臨床通過轉(zhuǎn)錄因子途徑治療哮喘提供新的理論依據(jù)。(3)地塞米松可能是通過下調(diào)BATF、RORγt的表達而降低Th17細胞的分化發(fā)育及分泌功能,進而減輕氣道的炎癥性損害及哮喘癥狀,最終達到抑制氣道炎癥的效果。
[Abstract]:Objective: To investigate the regulation mechanism of B cell activating transcription factor (BATF) on airway inflammation in acute asthmatic mice induced by ovalbumin (OVA) and its relationship with the orphan retinoic receptor gamma t (RORyt). Methods: (1) 24 healthy female BALB/c mice (SPF grade, body weight, 68 weeks) were randomly divided into 3. Group 0.9%, 0.9% saline control group (group NS), asthma model group (group AS), dexamethasone group (group DEX), 8.AS groups in each group. Group DEX mice were sensitized by intraperitoneal injection of ovalbumin antigen (100 mu g OVA and LMG hydroxide mixed suspension of sodium hydroxide) 200 UL, and NS group with isovolumetric saline in the same way Instead of antigenic fluid. Twenty-first days from twenty-first days, group NS was stimulated with physiological saline, AS group, and group DEX with 2% OVA ultrasonic atomization inhalation, 1 times a day, 40 minutes each time, 30 minutes before each stimulation for 5 days in.DEX group, and dexamethasone 1. Omg/kg intraperitoneal injection, NS group, AS group were given equal volume of saline intraperitoneal injection, time the same. The group of mice at the end of 24h was injected with intraperitoneal injection, and bronchoalveolar lavage and lung tissue were collected. (2) the count of leukocyte (WBC) and eosinophil (EOS), neutrophils (PMN) in bronchoalveolar lavage fluid (Bronchoalveolar lavage fluid, BALF), and double antibody sandwich enzyme-linked immunosorbent assay (ELISA) examination, respectively. The expression of interleukin 17 (Interleukin 17, IL-17) in the supernatant, pathological sections of lung tissue and HE staining were used to observe the degree of inflammatory infiltration in bronchopulmonary tissues, and the real-time fluorescence quantitative polymerase chain reaction (real time Polymerase Chain Reaction, RT-PCR) was used to detect the expression of IL-17, BATF, ROR gamma at the level of lung tissue. SPSS17.0 software was used to make statistical analysis on the experimental data of each group. The variables were expressed by mean mean deviation (x + s). The inter group variables were analyzed by single factor variance. The correlation of the two variables was analyzed with linear correlation. The statistical differences were all defined by P0.05. Results: (1) the HE staining results of pathological sections showed that the bronchial tube wall of group NS mice The smooth muscle was thin, smooth and smooth and regular. There was no secretions in the cavity. The airway mucosa folds were low and regular, the alveolar septum was wide and clear. There was no obvious inflammatory cell infiltration in the peribronchovascular and alveolar septum. The smooth muscle of the bronchial tube wall in the AS group was obviously thickened, a large number of mucous suppositories in the endotracheal cavity and the hypertrophy of the airway mucosa fold increased. Irregular birth, disorder of cilia, severe stenosis of bronchial tube, fracture of airway epithelium, incomplete exfoliation, infiltration of inflammatory cells in peribronchovascular tube and alveolar septum, thickening of alveolar septum, unclear boundary, EOS and P, and thickening of bronchial tube wall in group DEX and the thickening of bronchial tube wall in group of mice. The degree of narrowing of the lumen was less than that in the AS group, the secretion in the lumen decreased significantly, the alveolar septum was clearly visible, and the infiltration of inflammatory cells in the bronchovascular tube and the alveolar septum was improved. (2) the number of white blood cells in the AS group and the number of eosinophils and neutrophils in the BALF group were significantly increased (PO.01); compared with the AS group, the DEX group was compared with the group AS. The number of white blood cells, eosinophils and neutrophils decreased significantly (P0.01), but still more than group NS (P0.05). (3) ELISA results showed that the concentration of IL 1 17 protein in AS group BALF was significantly higher than that of NS group and DEX group (P0.05). The expression level of IL-17, BATF, ROR gamma t mRNA in the lung tissue of AS group was significantly higher than that of the NS group, and the difference in DEX group was statistically significant (PO.05). Compared with the AS group, the expression level of lung tissue in DEX group was reduced. (5) correlation Analysis: expression of lung tissue in mice The expression of ROR gamma t is positively correlated with WBC, EOS, PMN count in BALF (r=0.934,2=0.776, r=0.833,2=0.852,2=0.916, P are 0.05), and there is a significant positive correlation between BATF and ROR t in mice lung tissue. Conclusion: (1) in bronchial and lung tissues of acute asthmatic mice. The expression of BATF, IL-17, ROR gamma t was up-regulated and the expression of the three was positively correlated. (2) the increase of BATF expression may be the key factor in the airway inflammation in asthmatic mice, which provides a new theoretical basis for the treatment of asthma through the transcription factor pathway. (3) dexamethasone may reduce the fraction of Th17 cells by reducing the expression of BATF and ROR gamma t. The development and secretion function can reduce airway inflammation and asthma symptoms, and finally achieve the effect of inhibiting airway inflammation.
【學位授予單位】：瀘州醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R562.25

【參考文獻】

中國期刊全文數(shù)據(jù)庫 前1條

1 魏東;劉學東;;支氣管肺泡灌洗術(shù)治療難治性重度哮喘的臨床分析[J];中國醫(yī)藥科學;2011年15期

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本文編號：2107517