脂氧素A4对脂多糖攻击的大鼠肺泡Ⅱ型上皮细胞水通道蛋白5的影响  

Effect of lipoxin - A4 on the expression of aquaporin - 5 in rat alveolar type Ⅱ cells treated withlipopolysaccharide

作　　者：金胜威[1] 梅虹霞[1] 连庆泉[1] 

机构地区：[1]温州医学院附属第二医院麻醉科,浙江325000

出　　处：《中国急救医学》2009年第1期38-41,F0003,共5页Chinese Journal of Critical Care Medicine

基　　金：国家自然科学基金资助项目（No.30772088）;浙江省卫生高层次创新人才培养工程项目

摘　　要：目的探讨脂氧素A4（LipoxinA4，LXA4）对脂多糖（lipopolysaccharide，LPS）攻击的大鼠肺泡Ⅱ型上皮细胞（alveolar type Ⅱ penumonoeyte，ATⅡ）水通道蛋白（aquapofin，AQP）5的影响。方法对大鼠ATⅡ细胞进行分离、纯化，以1μg／mL的LPS刺激ATⅡ4h建立LPS攻击ATⅡ细胞模型。将ATⅡ随机分为：空白对照（无血清的培养基不含任何药物）组；溶剂对照（乙醇，0．7μL／mL）组；LPS（1μg／mL）组；LXA4（1×10^-7mol／mL）组；LPS＋LXA4组。用逆转录聚合酶链反应法（RT-PCR）检测大鼠ATⅡ中AQP5的信使核糖核酸（mRNA）的变化，免疫组织化学方法（IHC）检测ATⅡ中AQP5蛋白表达。结果1μg／mL的LPS刺激ATⅡ4h后，ATⅡ中AQP5的mRNA和蛋白表达较空白组明显减低（P〈0．01），而LPS＋LXA4组ATⅡ中AQP5的mRNA和蛋白表达较LPS组明显增高（P〈0．01），且LXA4组ATⅡ中AQP5的mRNA和蛋白表达较空白组也明显增高（P〈0．01）。结论大鼠ATⅡ上表达有AQP5，LXA4能促进AQP5mRNA和蛋白表达上调，提示LXA4可能通过上调AQP5的表达起到促进肺泡水肿液清除作用。Objective To study the effect of Lipoxins A4 （ LXA4 ） on the expression of aquaporin （AQP） 5 in type Ⅱ penumonocyte （ AT Ⅱ ） of rat treated with lipopolysacch - aride （ LPS ）. Methods Type Ⅱ penumonocyte of rats were isolated and puritfied, and the changes in cellular uhrastructure were studied by electron microscope, type Ⅱ penumonocyte were treated with 1 μg/mL LPS and divided randomly into five groups ： control group ; vehiculum （ alcohol, 0.7 μL/mL） ; endotoxin （ LPS, 1 μg/mL ） group; LXA4（1 ×10^-7mol/mL） group and LX A4 ＋ LPS group. AT Ⅱ was used for aquaporin（AQP）5 mRNA by reversal transcription poly chain reaction （ RT - PCR ） , and the expression of AQP5 protein was detected by immunohistochemistry （ IHC）. Results RT - PCR and immunohistochemistry showed that 1μg/mL LPS treatment of AT Ⅱ after 4 hours, the mRNA and protein expression levels of AQP5 were significantly descended in LPS group （P 〈 0.01 vs control group ） , However, up -regulation of the mRNA and protein expression levels of AQP5 by application of LXA4 in the model of AT Ⅱ treated with LPS（ P 〈 0.01 vs LPS group） , and the mRNA and protein expression levels of AQP5 were significantly elevated in LXA4 group contrast control group { P 〈 0. 01 ）. Conclusion AQP5 existed in AT Ⅱ of the rats , the LXA4 can up - regulate the mRNA and protein expression levels of AQP5 in AT II treated with LPS, and these findings suggest that LXA4 provided positive role to clear alveolar edema fluid in LPS - induced lung injury, and the role is likely due to up - regulate the expression of AQP5.

关 键 词：脂氧素A4 水通道蛋白5 肺泡Ⅱ型上皮细胞 脂多糖 

分 类 号：R332[医药卫生—人体生理学] R563[医药卫生—基础医学]