低渗液体外培养对气道上皮黏液分泌的影响及与水通道蛋白-5表达的相关性研究  

Study of mucus secretion and aquaporin - 5 expression of bronchial epithelium cultured in hypotonic medium

作　　者：闫霞[1] 曹官铭[1] 王晓龙[2] 周向东[2] 

机构地区：[1]重庆市急救医疗中心呼吸科,400014 [2]重庆医科大学附属第二临床学院呼吸科,400010

出　　处：《中国危重病急救医学》2007年第4期214-216,共3页Chinese Critical Care Medicine

摘　　要：目的 探讨体外气液界面培养气道上皮在低渗液条件下黏液分泌与水通道蛋白-5（AQP5）表达间的关系，以进一步阐明渗透浓度、AQP5在慢性气道黏液高分泌过程中的地位和作用。方法利用体外气液界面细胞培养模型，进行兔气道上皮细胞原代培养。培养1周后，培养液渗透浓度换为235mmol／L（实验A组）、255mmol／L（实验B组）、270mmol／L（实验C组）和282mmol／L（对照组）。培养12h后，采用逆转录-聚合酶链反应（RT—PCR）检测各组AQP5mRNA表达，采用蛋白质免疫印迹法检测上清液中黏蛋白5AC（MUC5AC）分泌量。结果 倒置显微镜下各培养组细胞成多层生长。各实验组中MUC5AC分泌量均高于对照组（P〈0．001），其中以实验A组为最高；而AQP5 mRNA均明显低于对照组（P〈0．001），以实验A组为最低；实验组AQP5mRNA表达与MUC5AC呈显著负相关（r=-0.77，P〈0.001）。结论 在气液界面细胞培养模型中，低渗液条件下MUC5AC分泌量显著增加，较好地模拟了慢性气道炎症黏液高分泌的体内环境；AQP5mRNA表达与MUC5AC呈显著负相关。低渗环境以及由此引起的AQP5mRNA表达可能参与慢性阻塞性肺疾病黏液高分泌的形成过程。Objective To study mucus secretion and aquaporin - 5 （AQPS） expression of bronchial epithelium cultured at the air - liquid interface of hypotonic medium, and to elucidate the role of AQP5 and osmotic pressure in airway mucus hypersecretion process. Methods With the air - liquid interface culture model, rabbit tracheal epithelium was cultured at the air - liquid interface. One week after culture, media of 235, 255 and 270 mmol/L were used for experimental groups, and that of 282 mmol/L for culture in control group. Twelve hours after the culture, reverse transcriptase polymerase chain reaction （RT -PCR） was used to detect AQP5 mRNA in each experimental group, and Western blotting to determine mucin5AC （MUC5AC） in the supernatant. The same procedures were done in control group. Results Multilayers of cultured cells were observed with inverted microscope in each group. MUC5AC expressions in experimental groups were significantly higher, whereas AQP5 mRNA levels were obviously lower compared with those in control group （all P 〈 0.001）, the changes in 235 mmol/L group were most obvious. Moreover, AQP5 mRNA and MUCSAC in experimental groups were notably negatively correlated （r=- 0. 77, P〈 0. 001）. Conclusions The model of air - liquid interface culture could increase MUCSAC expression, which provides an environment closely resembles that in the body is an ideal framework for the research of mucus hypersecretion of chronic obstructive pulmonary disease （COPD）. Expressions of AQP5 mRNA and MUC5AC are negatively correlated. Hypotonicity and reduction of AQP5 mRNA may play critical roles in airway mucus secretion process in COPD.

关 键 词：水通道蛋白 黏蛋白 气液界面 低渗液 

分 类 号：R686[医药卫生—骨科学]