人胸膜间皮细胞水通道蛋白1～10 mRNA的表达  

The Expression of AQP_(1～10) mRNA in The Human Mesothelial Cells

作　　者：王洪峰[1] 刘宏博[1] 曲文秀[1] 何平[1] 王志云 李胜岐[1] 

机构地区：[1]中国医科大学附属第二医院(盛京医院)呼吸内科,沈阳110004 [2]河南省鹤壁肿瘤医院胸外科,鹤壁458000

出　　处：《生物化学与生物物理进展》2007年第1期57-62,共6页Progress In Biochemistry and Biophysics

摘　　要：体外培养人胸膜间皮细胞(HPMC),检测人胸膜间皮细胞水通道蛋白1～10mRNA的表达,探讨其在胸腔内液体平衡中的意义.从胸腔积液中分离人胸膜间皮细胞,进行培养,用形态学和免疫组化染色进行细胞鉴定.用RT-PCR检测水通道蛋白1～10(AQP1～10)mRNA在人胸膜间皮细胞上的表达.成功建立人胸膜间皮细胞体外培养模型,鉴定证实为间皮细胞,人胸膜间皮细胞上AQP1～10mRNA均有表达,AQP1、AQP9、AQP10表达丰富.人胸膜间皮细胞存在AQP1～10mRNA的表达,结合已知水通道蛋白的功能,证实人胸膜间皮细胞参于胸腔内液体转运.Aquaporins （AQP） are integral membrane proteins that serve as channels in the transfer of water, and in some cases, small solutes across the membrane. In order to detect the expression of AQP1-10 mRNA in the human pleural mesothelial cells, culture reproducible model of human pleural mesothelial cells was established in vitro. The value of aquaporin water channels was investigated in pleural fluid dynamics. Mesothelial cells were isolated from pleural effusion fluid of nonmalignant pleural effusion patients and cultured in vitro. The mesothelial cells were identified by morphology and streptomyces protein-peroxidase（S-P） procedure. RT-PCR was used to detect the expression of AQPl-10 mRNA in human pleural mesothelial cells. Establishment of culture reproducible model achieved success of human pleural mesothelial cells in vitro. Immunocytochemistry revealed that cytokeratin and vimentin expressed＂ positive, V Ⅲ factor associated antigen and CD45 was negtive. Confluent human pleural mesothelial cells appeared multipolar and like cabblestone.Under electron microscopy numerous surface microvilli and abundant endoplasmic reticulum were observed. The cells was identified that it was mesothelial cells. The cells isolated from human pleural effusion fluid have higher purity quotient. RT-PCR studies showed that all of the AQP1-1O mRNA expressed in mesothelial cells. AQPI, AQP9, AQPIo mRNA have abundant expression in mesothelial cells and significant deviation was found between AQP1, AQP9, AQP10 and AQP2, AQP3, AQP7, AQP8- Human pleural mesothelial cells expressed AQP1-10 mRNA, all of them have relation with the function of human pleural mesothelial cells. It was confirmed that mesothelial cells have important contribution in pleural fluid dynamics. Aquaporins （AQP） are involved in rapid and active gating of water across biological membranes. Expression of aquaporins was intervened, new treat method may be found ofpleural effusion.

关 键 词：人胸膜间皮细胞 培养 水通道蛋白 

分 类 号：R332.2[医药卫生—人体生理学]