乙酰唑胺对大鼠肾脏近曲小管上皮细胞作用的差异蛋白分析及其与AQP1抑制的关系  

Differential protein analysis in rat renal proximal tubule epithelial cells in response to acetazolamide and its relation with the inhibition of AQP1

作　　者：母生梅[1] 暨荀鹤[1] 马兵[1] 于和鸣 李学军[1] 

机构地区：[1]北京大学医学部基础医学院药理学系,北京100083 [2]国家计划生育委员会科学研究所,北京100081

出　　处：《药学学报》2003年第3期169-172,共4页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金 (3 9770 2 863 9770 7753 0 1710 90);美国洛克菲勒基金 (RF93 0 63 #83 )

摘　　要：目的　研究乙酰唑胺抑制大鼠肾脏近曲小管上皮细胞水孔蛋白 1(aquaporin 1,AQP1)表达的内源性机制。方法　将原代培养的大鼠肾脏近曲小管上皮细胞分为两组 ,一组给予 1× 10 - 5mol·L- 1 乙酰唑胺 ,另一组为阴性对照。待细胞长至亚融合状态时裂解细胞 ,用等电聚焦电泳对裂解物进行差异蛋白筛选 ,并对差异蛋白作肽质量指纹谱鉴定。结果　给乙酰唑胺后细胞中出现两个差异蛋白 ,其中等电点 (pI)为 3 8的蛋白给药后的表达减少 ,数据库检索发现其氨基酸序列与来源于大鼠肝脏和睾丸的刷状缘肌球蛋白有 2 1 4 %的相似性 ;而与来源于大鼠脑的糖原磷酸化酶有 2 7%相似性。另外一条pI为 5 5的蛋白被诱导增多 ,它与来源于大鼠肺脏的α亚型磷脂酰肌醇转移蛋白的相似性为 2 0 %。结论　乙酰唑胺可能通过影响pI 3 8蛋白或pI5Aim To study the endogenous mechanism for the inhibition of aquaporin 1 expression in rat renal proximal tubule epithelial cells in response to acetazolamide. Methods Primary cultured rat renal proximal tubule epithelia cells were divided into two groups: one was subjected to 1×10 -5 mol·L -1 acetazolamide, the other served as normal control. When grown to sub confluency, the cells were disintegrated to perform isoelectrofocusing electrophoresis in order to find the differential proteins induced by the acetazolamide treatment. The differential proteins were defined by peptide mass fingerprinting technology. Results Two differential proteins were found in the cell disintegrant. The pI 3 8 protein was reduced after treatment, which showed 21 4% similarity with the brush border membrane myosin from rat brain and testis, and 27% with glycogen phosphorylase; The pI 5 5 protein was increased on the contrary, with 20% similarity to phosphatidylinositol transfer protein alpha isoform. Conclusion Acetazolamide inhibited AQP1 expression probably by affecting the expression of pI 3 8 and pI 5 5 proteins.

关 键 词：乙酰唑胺 水孔蛋白 肾脏近曲小管上皮细胞 肽质量指纹谱 

分 类 号：R977.7[医药卫生—药品] R978.2[医药卫生—药学]