摘要
目的探讨脂氧素A4（LipoxinA4，LXA4）对脂多糖（hpopolysaccharide，LPS）攻击的大鼠肺泡Ⅱ型上皮细胞（type Ⅱpenumonocyte，ATⅡ）水通道蛋白（aquaporin，AQP）1，3，5的影响。方法每次取SPF级健康SD雄性大鼠，体质量200～250g一只，对大鼠肺泡Ⅱ型上皮细胞进行分离、纯化，鉴定，得到纯度为≥90％的肺泡Ⅱ型上皮细胞，将细胞随机分为：①溶剂对照（乙醇，0．7μL／mL）组；②空白组（无血清的培养基不含任何药物）；③LXA4（1×10^-7mol／mL）组；④LPS（1μg／mL）组；⑤LPS＋LXA4（1μG／mL LPS＋1×10^-7mol／mL LXA4）组。药物刺激4h后用逆转录聚合酶链反应法（RT-PCR）检测大鼠肺泡Ⅱ型上皮细胞上AQP1，AQP3和AQP5的mRNA的表达，免疫组织化学方法（IHC）检测肺泡Ⅱ型上皮细胞上AQP1，AQP3和AQP5蛋白的表达。试验重复6次。采用方差分析进行统计学处理。结果RT-PCR和免疫组织化学法结果显示，用1μg/mL的LPS刺激肺泡Ⅱ型上皮细胞4h后ATⅡ上AQP1，AQP3和AQP5的mRNA和蛋白表达较空白组明显减低（P〈0．01），而LPS＋LXA4组中ATⅡ上AQP1，AQP3和AQP5的mRNA和蛋白表达较LPS模型组有明显增高（LPS＋LXA4，AQP1：0.647±0．132，AQP3：0．900±0．856，AQP5：0．879±0．058；LPS，AQP1：0．297±0．133，AQP3：0．512±0．113，AQP5：0．647±0．110；P〈0．01），且LXA4组中ATⅡ上AQP1，AQP3和AQP5的mRNA和蛋白表达较空白组也有明显增高（LXA4，AQP1：0．539±0．142，AQP3：0．818±0．176，AQP5：0．841±0．066；Blank Control，AQP1：0．518±0．139；AQP3：0．138±0．136，AQP5：0．755±0．066；P〈0．01）。结论大鼠肺泡Ⅱ型上皮细胞上表达有AQP1，AQP3和AQP5，LXA4能促进脂多糖攻击的肺泡Ⅱ型上皮细胞上AQP1，AQP3和AQP5mRNA和蛋白表达上调。
Objective To study the effects of Lipoxins A4（LXA4） on the expressions of aquaporin （AQP）1, 3, 5 in type Ⅱ pneumonocytes （AT Ⅱ ） of rat treated with lipopolysaccharide （LPS）. Method One pathogenfree male Sprague Dawley（SD）rat every time, weighing 200 - 250 g, were used for the study. The type Ⅱ penumonocytes of rats were isolated and purified, and the changes of cellular ultrastructure were observed by electron mieroscopo in order to get the purity quotient 〉 90% .The type Ⅱ pneumonocytes were divided randomly into five groups, namely, vehiculum group （alcohol 0.7 μL/mL）, control group, LXA4 group （ 1 × 10^- 7 mol/mL）, endotoxin group （LPS 1 μg/mL） and LX A4 ＋ LPS group （LXA4 1 ×10^-7 moL/mL,LPS 1 μg/mL）. AQP-1,3,5 mRNA of in the type Ⅱ penumonecytes were assayed by using reversal transcription poly chain reaction （RT-PCR）, and the expressions of AQP-1, 3, 5 protein were detected by using immunohistochemistry （IHC）. One each specimen, these tests were repeated for six times. ANOVA was used for statistical analysis. Results RT-PCR and IHC showed that when AT Ⅱ treated with 1 μg/mL LPS for 4 hours, the AQP-1, 3, 5 mRNA and the expressions of AQP-1, 3, 5 protein were significantly decreased in LPS group compared with control group （ P 〈 0.01）. However,the AQP-1, 3, 5 mRNA and the expressions of AQP-1, 3, 5 protein after application of LXA4 significantly increased in LPS ＋ LXA4 group in comparison with LPS group （LPS ＋ LXA4, AQP1 ： 0.647 ± 0.132, AQP3 ： 0.900 ±0.856,AQP5：0.879 ± 0.058; LPS, AQP1 ： 0.297 ± 0.133, AQP3：0.512 ± 0.113, AQP5：0.647 ± 0.110; P 〈 0.01）. The AQP-1, 3, 5 rnRNA and the expressions of AQP-1, 3, 5 protein were signitieantly increased in LXA4 group in comparison with control group （ LXA4, AQP1 ： 0.539 ± 0.142, AQP3：0.818 ±0. 176, AQP5：0.841 ± 0.066; Blank Control,AQP1：0.518 ±0.139 ; AQP3 ： 0.138 ± 0.136,AQP5：0.766 ± 0.066; P 〈 0.01 ）. Conclusions AQP-1, 3, 5 exist in type Ⅱ penumonocyte of rats, and the LXA4 can upegulate the mRNA and protein expressions of AQP-1, 3, 5 in Type Ⅱ penumonocytes of rats treated with LPS.
出处
《中华急诊医学杂志》
CAS
CSCD
北大核心
2009年第4期406-411,共6页
Chinese Journal of Emergency Medicine
基金
基金项目：国家自然科学基金资助项目（30772088）
浙江省卫生高层次创新人才培养工程项目
关键词
脂氧素A4
水通道蛋白
肺泡Ⅱ型上皮细胞
脂多糖
Lipoxins A4 （LXA4）
Aquaporins（AQPs）
Alveolar type Ⅱ cells
Lipopolysaccharide （ LPS ）
