發(fā)布時間：2018-04-05 08:57

  本文選題：哮喘　切入點：肺泡巨噬細胞　出處：《遵義醫(yī)學院》2017年碩士論文

【摘要】：目的:探討白細胞介素-31對哮喘小鼠肺泡巨噬細胞表達CC趨化因子配體2的影響。方法:將雌性BALB/c小鼠隨機分為哮喘組(AS)和生理鹽水對照組(NS),每組10只。兩組小鼠于第1、13天分別以O(shè)VA和生理鹽水腹腔聯(lián)合皮下注射致敏,第19d分別予10%OVA溶液、生理鹽水連續(xù)霧化激發(fā)5d,末次激發(fā)24h后處死小鼠。測定小鼠BALF細胞總數(shù)和細胞分類計數(shù),HE染色觀察肺組織病理學改變。體外對小鼠行支氣管肺泡灌洗獲得BALF,采用貼壁法獲得肺泡巨噬細胞。觀察細胞形態(tài),體外培養(yǎng)至12-24小時,采用臺盼藍染色鑒定細胞活性,瑞氏-吉姆薩染色鑒定細胞形態(tài),采用流式細胞術(shù)鑒定細胞表型特征。NS組和AS組小鼠處死后體外獲得相應(yīng)組的肺泡巨噬細胞,將不同濃度(10ng/ml、50ng/ml、100ng/ml)的白細胞介素-31加入到含有相同肺泡巨噬細胞數(shù)量的不同組別培養(yǎng)基中,分別于加樣后12h、24h、36h行實時熒光定量PCR(RT-PCR)檢測CC趨化因子配體2的表達水平。結(jié)果:AS組小鼠BALF細胞總數(shù)、細胞分類計數(shù)、氣道炎癥細胞浸潤評分、氣道壁厚度、均高于NS組。肺泡巨噬細胞呈典型貼壁生長,細胞呈圓形、橢圓形及長梭形。體外分離肺泡巨噬細胞存活率≥95%,純度≥95%。肺泡巨噬細胞雙表達CD11c、F4/80。在IL-31干預(yù)下AS組肺泡巨噬細胞CCL2的表達量大于NS組。CCL2的表達量與IL-31干預(yù)濃度呈正相關(guān)。相同濃度下,CCL2的表達量與IL-31干預(yù)時間呈正相關(guān)。IL-31可促進哮喘小鼠肺泡巨噬細胞表達CCL2,CCL2的表達量隨IL-31干預(yù)濃度及時間呈增加趨勢。結(jié)論:IL-31可促進哮喘小鼠肺泡巨噬細胞表達CCL2,隨著IL-31干預(yù)濃度的增加及時間的延長,CCL2表達量也隨其呈正相關(guān)。
[Abstract]:Aim: to investigate the effect of interleukin-31 (IL-31) on the expression of CC chemokine ligand 2 in alveolar macrophages of asthmatic mice.Methods: female BALB/c mice were randomly divided into asthma group (n = 10) and saline control group (n = 10).The mice in the two groups were sensitized by intraperitoneal injection of OVA and normal saline on the 1st day of the 13th day, respectively. On the 19th day, the mice were treated with 10%OVA solution for 5 days. The mice were killed after the last stimulation for 24 hours.The total number of BALF cells and cell classification in mice were measured and the pathological changes of lung tissue were observed by HE staining.BALF was obtained by bronchoalveolar lavage in vitro and alveolar macrophages were obtained by adherent method.Cell morphology was observed and cultured for 12 to 24 hours in vitro. Trypan blue staining was used to identify cell activity, and Rayleigh Jimsa staining was used to identify cell morphology.The phenotypic characteristics of the cells were identified by flow cytometry. The corresponding alveolar macrophages were obtained after the mice of NS and as groups were killed in vitro.Results the total number of BALF cells, cell classification count, airway inflammatory cell infiltration score and airway wall thickness in as group were higher than those in NS group.Alveolar macrophages were typical adherent to the wall, and the cells were round, oval and fusiform.The survival rate of alveolar macrophages in vitro was 鈮，

本文編號：1714063