内毒素致大鼠肾髓质上皮细胞损伤与水通道蛋白2的表达  

Relationship between Aquaporin2 and LPS-induced Injury of Mouse Inner Medullary Collecting Duct Cells

作　　者：何静妮[1] 姚旭[1] 王勇[1] 周勇[1] 刘金钢[1] 

机构地区：[1]中国医科大学附属盛京医院胆道疝减肥外科病房,沈阳110004

出　　处：《中国医科大学学报》2010年第10期819-822,共4页Journal of China Medical University

基　　金：辽宁省教育厅高校科研基金资助项目(2004D282)

摘　　要：目的观察内毒素(LPS)对大鼠肾髓质上皮细胞mIMCD-3的影响及水通道蛋白2(AQP2)表达的变化。方法免疫组化检测AQP2在mIMCD-3的表达及定位。LPS与mIMCD-3培养不同时间后用CCK-8检测吸光度,计算细胞抑制率,检测凋亡和AQP2的表达情况,并做光密度分析。结果免疫组化可见,细胞质中大量棕红色颗粒,证明mIMCD-3表达AQP2。LPS作用于细胞检测细胞抑制率,不同浓度、不同时间组分别做单因素方差分析,具有统计学差异(P<0.05)。将10μg/L的LPS溶液作用于mIMCD-30h,4h,24h和48h后,经流式细胞仪检测结果表明,LPS对mIMCD-3的抑制作用主要表现为细胞凋亡,随作用时间的延长,细胞凋亡增加(P<0.01),作用时间和凋亡率具有直线相关性(R=0.9920)。同时行免疫组化可见,细胞中棕黄色颗粒变稀疏,AQP2表达下调。平均光密度分析具有统计学差异(P<0.01)。结论 mIMCD-3表达AQP2,基础状态下主要定位于细胞浆中。LPS抑制mIMCD-3增殖,具有时间依赖性和剂量依赖性。LPS对细胞增殖的抑制作用主要表现为诱导细胞凋亡。随作用时间延长,细胞凋亡率增加,两者具有直线相关。LPS介导的肾髓质上皮细胞凋亡通路中,AQP2起重要作用。Objective To investigate the effect of endotoxin（LPS）on mouse inner medullary collecting duct cells（mIMCD-3）and the corresponding changes of aquaporin 2（AQP2）.Methods Immunohistochemical staining was used to detect the expression and localization of AQP2 in mIMCD-3.CCK-8 was used to determine the inhibitory effect after incubating with LPS for 0 to 72 hours.Flow cytometry was employed to calculate the inhibitory rate.And the change of AQP2 was detected by immunohistochemistry.Results AQP2 immunohistochemical staining result showed a large number of brown-red particles piled up like plates could be seen in the cytoplasm.As mIMCD-3 incubated with different concentrations of LPS for different time,significant differences were found （P 〈 0.05）.Flow cytometry results showed that the inhibitory effect of LPS mainly showed as apoptosis.The apoptosis rate increased time-dependently（P 〈 0.01）,and there was a linear correlation between the apoptosis rate and the time （R=0.9920）.The results of immunochemical staining showed,under the treatment of LPS,the expression of AQP2 were downregulated （P 〈 0.01）.Conclusion mIMCD-3 expresses AQP2 which mainly localizes in the cytoplasm.LPS induces the apoptosis and inhibits the proliferation of mIMCD-3 with a time-dependent and dose-dependent manner.AQP2 may play an important role in LPS-induced apoptosis of mIMCD-3.

关 键 词：大鼠肾髓质上皮细胞 内毒素 水通道蛋白2 CCK-8 流式细胞法 

分 类 号：R361.3[医药卫生—病理学]