發(fā)布時(shí)間：2018-06-13 18:14

  本文選題：B細(xì)胞活化轉(zhuǎn)錄因子 + ROR�。� 參考：《瀘州醫(yī)學(xué)院》2013年碩士論文

【摘要】：目的：觀察B細(xì)胞活化轉(zhuǎn)錄因子(B-cell activiting transcription factor, BATF)、維甲酸相關(guān)孤核受體(retinoic acid-related orphan recepptor γt, ROR γ t白細(xì)胞介素-17(interleukin-17, IL-17)在哮喘小鼠脾臟組織中的表達(dá),探討B(tài)ATF在哮喘發(fā)病機(jī)制中的作用,為哮喘治療尋找新途徑。方法：取健康雌性BALB/c小鼠15只,隨機(jī)分為正常對(duì)照(NS)組、哮喘(AS)組、地塞米松(DXM)組,每組小鼠均為5只。AS組、DXM組小鼠分別于第0、7、14天用卵清蛋白(OVA)100ug與氫氧化鋁(Al (OH) 3) 1mg混合的生理鹽水混懸液0.2ml腹腔內(nèi)注射致敏,NS組則用等量生理鹽水代替行腹腔注射；第21天開(kāi)始NS組予以生理鹽水,AS組及DXM組予1%的OVA生理鹽水溶液行超聲霧化吸入,每天1次,每次40分鐘,連續(xù)5天進(jìn)行激發(fā)；DXM組于每次激發(fā)前30分鐘,予地塞米松磷酸鈉注射液1.0mg/kg腹腔注射,NS組及AS組則予以等量的生理鹽水腹腔注射,時(shí)間同上。所有小鼠都于末次激發(fā)24小時(shí)后處死。收集支氣管肺泡灌洗液(BALF)行白細(xì)胞計(jì)數(shù)及分類(lèi)計(jì)數(shù)；肺組織病理切片HE染色來(lái)評(píng)估小鼠氣道及肺組織炎性細(xì)胞浸潤(rùn)情況；酶聯(lián)免疫吸附法(ELISA)檢測(cè)脾臟組織勻漿IL-17的濃度；real-time PCR法檢測(cè)脾臟組織中BATF mRNA、ROR γ t mRNA的表達(dá)水平。結(jié)果：(1)肺組織病理學(xué)HE染色：AS組可見(jiàn)小氣道及小血管周?chē)罅康难仔约?xì)胞浸潤(rùn),粘膜及粘膜下層水腫,肺泡腔內(nèi)可見(jiàn)嗜酸性粒細(xì)胞、中性粒細(xì)胞、單核細(xì)胞及溢出的紅細(xì)胞；DXM組炎細(xì)胞、滲出物較AS組少,支氣管上皮基本完整,可見(jiàn)明顯的炎癥吸收；NS組小氣道粘膜無(wú)明顯水腫,管腔光滑、無(wú)閉塞,周?chē)⊙芑疚匆?jiàn)炎性細(xì)胞的浸潤(rùn)。(2)與NS組比較,AS組、DXM組小鼠BALF中白細(xì)胞總數(shù)、中性粒細(xì)胞數(shù)和嗜酸性粒細(xì)胞數(shù)明顯增多(P0.05),與AS組比較,DXM組BALF中白細(xì)胞總數(shù)、中性粒細(xì)胞數(shù)和嗜酸性粒細(xì)胞數(shù)有明顯降低(P0.05)。(3)脾臟組織勻漿IL-17的濃度：AS組IL-17濃度最高,DXM組其次,NS組最低,三組之間的差異有統(tǒng)計(jì)學(xué)意義(P0.05),且與NS組比較,AS組及DXM組IL-17濃度均明顯增高,差異有統(tǒng)計(jì)學(xué)意義(P0.05),DXM組較AS組IL-17濃度降低,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。(4)脾臟組織中BATF mRNA.ROR γ t mRNA表達(dá)水平：AS組水平最高,DXM組其次,NS組最低,三組之間的差異有統(tǒng)計(jì)學(xué)意義(P0.05),且與NS組比較,AS組及DXM組BATF mRNA. ROR γ t mR NA表達(dá)水平均明顯增高,差異有統(tǒng)計(jì)學(xué)意義(P0.05),DXM組較AS組BATF mRNA、ROR y t mRNA表達(dá)水平降低,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。(5)相關(guān)分析表明：脾臟組織中IL-17的濃度與BALF中白細(xì)胞總數(shù)、中性粒細(xì)胞數(shù)、嗜酸性粒細(xì)胞數(shù)成正相關(guān)(r=0.838,0.737,0.882,P均0.005)；脾臟組織BATF mRNA的表達(dá)量與ROR γ t mRNA表達(dá)量、脾組織勻漿IL-17濃度呈正相關(guān)(r=0.807,0.833,P均0.005)：脾組織勻漿IL-17濃度與脾臟組織ROR γ t mRNA表達(dá)量呈正相關(guān),(r=0.890,P0.005)。結(jié)論：(1)BATF與ROR γ t共同調(diào)節(jié)IL-17的分泌,并通過(guò)IL-17的致炎作用參與哮喘氣道炎癥的發(fā)生發(fā)展,(2)地塞米松可能通過(guò)BATF、ROR γ t途徑部分阻止脾源性Th17細(xì)胞的分化,減少I(mǎi)L-17的分泌,從而改善哮喘氣道炎癥細(xì)胞的浸潤(rùn),緩解哮喘癥狀。
[Abstract]:Objective: To observe the expression of B cell activation transcription factor (B-cell activiting transcription factor, BATF), retinoic acid related soliton receptor (retinoic acid-related orphan recepptor gamma T, ROR gamma) in the spleen tissues of asthmatic mice and to explore the role of asthma in the pathogenesis of asthma. Methods: 15 healthy female BALB/c mice were randomly divided into normal control (NS) group, asthma (AS) group and dexamethasone (DXM) group, each group was 5.AS groups, and DXM mice were injected intraperitoneally with the mixture of ovalbumin 100ug (OVA) 100ug and aluminum hydroxide (Al (OH) 3) on day 0,7,14. Group NS was injected with equal physiological saline instead of intraperitoneal injection; group NS was given physiological saline at twenty-first days, group AS and group DXM were given 1% OVA physiological saline solution by ultrasonic atomization inhalation, 1 times a day, 40 minutes each time for 5 days, and 30 minutes before each stimulation, group DXM was given to Dexamethasone Sodium Phosphate Injection 1.0mg/kg abdominal cavity. Injection, group NS and group AS were injected with equal amount of normal saline. All mice were killed after 24 hours of last excitation. Bronchoalveolar lavage fluid (BALF) was collected to count the white blood cell count and classified count; lung tissue pathological section HE staining was used to evaluate the infiltration of inflammatory cells in the airway and lung tissues; enzyme linked immunosorbent assay. The concentration of IL-17 in the spleen tissue homogenate was detected by adsorption method (ELISA); the expression level of BATF mRNA and ROR t mRNA in the spleen tissues was detected by real-time PCR. Results: (1) HE staining of lung histopathology: AS group could see large number of inflammatory cell infiltration in small airway and small vessels, mucous membrane and submucosa edema, and eosinophil in the alveoli. Cells, neutrophils, mononuclear cells and spillover red cells, group DXM inflammatory cells, exudates less than group AS, bronchial epithelium was basically complete, visible inflammatory absorption, NS group small airway mucosa without obvious edema, smooth lumen, no occlusion, surrounding small vascular basically no infiltration of inflammatory cells. (2) compared with group NS, AS group, group DXM mice B The total number of leucocytes and neutrophils and eosinophils in ALF increased significantly (P0.05). Compared with the AS group, the total number of white blood cells, neutrophils and eosinophils in group DXM were significantly decreased (P0.05). (3) the concentration of IL-17 in the spleen tissue homogenate was the highest in the AS group, the second in the DXM group, the lowest in the NS group and the difference between the three groups. The difference was statistically significant (P0.05), and compared with the NS group, the IL-17 concentration in AS and DXM groups increased significantly (P0.05), and the concentration of IL-17 in group DXM was lower than that in AS group, and the difference was statistically significant (P0.05). (4) the level of BATF mRNA.ROR gamma expression in the spleen tissue was the highest, followed by the group, the lowest, three groups. The difference was statistically significant (P0.05), and compared with the NS group, the BATF mRNA. ROR gamma t mR NA expression level in the group AS and the DXM group was significantly higher, and the difference was statistically significant (P0.05). (5) the correlation analysis showed that the concentration of the spleen in the spleen tissue and the concentration of the spleen T The total number of leucocytes, neutrophils and eosinophils were positively correlated (r=0.838,0.737,0.882, P 0.005); the expression of BATF mRNA in the spleen tissue was positively correlated with the ROR gamma t mRNA expression, and the concentration of IL-17 in the spleen tissue homogenate was positively correlated (r=0.807,0.833, P 0.005): the IL-17 concentration of spleen tissue homogenization and the expression of splenic tissue ROR gamma expression Cheng Zhengxiang Guan, (r=0.890, P0.005). Conclusion: (1) BATF and ROR gamma t regulate the secretion of IL-17, and participate in the development of airway inflammation through the inflammatory action of IL-17. (2) dexamethasone may pass BATF, ROR gamma t pathway partially prevent the differentiation of spleen derived Th17 cells, reduce the secretion of IL-17, and thus improve the infiltration of airway inflammatory cells in asthma. Relieve the symptoms of asthma.
【學(xué)位授予單位】：瀘州醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類(lèi)號(hào)】：R562.25

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相關(guān)期刊論文 前1條

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本文編號(hào)：2014948