本文選題：哮喘　切入點(diǎn)：肺泡巨噬細(xì)胞　出處：《遵義醫(yī)學(xué)院》2017年碩士論文

【摘要】：目的:探討白細(xì)胞介素-31對(duì)哮喘小鼠肺泡巨噬細(xì)胞表達(dá)CC趨化因子配體2的影響。方法:將雌性BALB/c小鼠隨機(jī)分為哮喘組(AS)和生理鹽水對(duì)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(cè)定小鼠BALF細(xì)胞總數(shù)和細(xì)胞分類計(jì)數(shù),HE染色觀察肺組織病理學(xué)改變。體外對(duì)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(xì)胞。觀察細(xì)胞形態(tài),體外培養(yǎng)至12-24小時(shí),采用臺(tái)盼藍(lán)染色鑒定細(xì)胞活性,瑞氏-吉姆薩染色鑒定細(xì)胞形態(tài),采用流式細(xì)胞術(shù)鑒定細(xì)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(xì)胞,將不同濃度(10ng/ml、50ng/ml、100ng/ml)的白細(xì)胞介素-31加入到含有相同肺泡巨噬細(xì)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(shí)時(shí)熒光定量PCR(RT-PCR)檢測(cè)CC趨化因子配體2的表達(dá)水平。結(jié)果:AS組小鼠BALF細(xì)胞總數(shù)、細(xì)胞分類計(jì)數(shù)、氣道炎癥細(xì)胞浸潤評(píng)分、氣道壁厚度、均高于NS組。肺泡巨噬細(xì)胞呈典型貼壁生長,細(xì)胞呈圓形、橢圓形及長梭形。體外分離肺泡巨噬細(xì)胞存活率≥95%,純度≥95%。肺泡巨噬細(xì)胞雙表達(dá)CD11c、F4/80。在IL-31干預(yù)下AS組肺泡巨噬細(xì)胞CCL2的表達(dá)量大于NS組。CCL2的表達(dá)量與IL-31干預(yù)濃度呈正相關(guān)。相同濃度下,CCL2的表達(dá)量與IL-31干預(yù)時(shí)間呈正相關(guān)。IL-31可促進(jìn)哮喘小鼠肺泡巨噬細(xì)胞表達(dá)CCL2,CCL2的表達(dá)量隨IL-31干預(yù)濃度及時(shí)間呈增加趨勢(shì)。結(jié)論:IL-31可促進(jìn)哮喘小鼠肺泡巨噬細(xì)胞表達(dá)CCL2,隨著IL-31干預(yù)濃度的增加及時(shí)間的延長,CCL2表達(dá)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(hào)：1714063