白细胞介素-31对哮喘小鼠肺泡巨噬细胞表达CC趋化因子配体2的影响

发布时间：2018-04-05 08:57

  本文选题：哮喘　切入点：肺泡巨噬细胞　出处：《遵义医学院》2017年硕士论文

【摘要】：目的:探讨白细胞介素-31对哮喘小鼠肺泡巨噬细胞表达CC趋化因子配体2的影响。方法:将雌性BALB/c小鼠随机分为哮喘组(AS)和生理盐水对照组(NS),每组10只。两组小鼠于第1、13天分别以OVA和生理盐水腹腔联合皮下注射致敏,第19d分别予10%OVA溶液、生理盐水连续雾化激发5d,末次激发24h后处死小鼠。测定小鼠BALF细胞总数和细胞分类计数,HE染色观察肺组织病理学改变。体外对小鼠行支气管肺泡灌洗获得BALF,采用贴壁法获得肺泡巨噬细胞。观察细胞形态,体外培养至12-24小时,采用台盼蓝染色鉴定细胞活性,瑞氏-吉姆萨染色鉴定细胞形态,采用流式细胞术鉴定细胞表型特征。NS组和AS组小鼠处死后体外获得相应组的肺泡巨噬细胞,将不同浓度(10ng/ml、50ng/ml、100ng/ml)的白细胞介素-31加入到含有相同肺泡巨噬细胞数量的不同组别培养基中,分别于加样后12h、24h、36h行实时荧光定量PCR(RT-PCR)检测CC趋化因子配体2的表达水平。结果:AS组小鼠BALF细胞总数、细胞分类计数、气道炎症细胞浸润评分、气道壁厚度、均高于NS组。肺泡巨噬细胞呈典型贴壁生长,细胞呈圆形、椭圆形及长梭形。体外分离肺泡巨噬细胞存活率≥95%,纯度≥95%。肺泡巨噬细胞双表达CD11c、F4/80。在IL-31干预下AS组肺泡巨噬细胞CCL2的表达量大于NS组。CCL2的表达量与IL-31干预浓度呈正相关。相同浓度下,CCL2的表达量与IL-31干预时间呈正相关。IL-31可促进哮喘小鼠肺泡巨噬细胞表达CCL2,CCL2的表达量随IL-31干预浓度及时间呈增加趋势。结论:IL-31可促进哮喘小鼠肺泡巨噬细胞表达CCL2,随着IL-31干预浓度的增加及时间的延长,CCL2表达量也随其呈正相关。
[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，

本文编号：1714063