Effect of E6, a novel calmodulin inhibotor, on activity of P-glycoprotein in purified primary cultured rat brain microvessel endothelial cells  

新钙调素抑制剂E6对纯化的原代培养大鼠脑微血管内皮细胞上P-糖蛋白活性的影响(英文)

作　　者：祝浩杰[1] 刘国卿[1] 

机构地区：[1]中国药科大学药理教研室

出　　处：《Acta Pharmacologica Sinica》2003年第11期1143-1149,1175,共8页中国药理学报（英文版）

基　　金：Project supported by the National Natural Science Foundation of China, № 39870885.

摘　　要：AIM: To study the effect of E6, a novel calmodulin inhibitor, on the activity of P-glycorprotein(P-gp) in purified primary cultured rat brain microvessel endothelial cells(RBMEC). METHODS: Flow cytometry was used to purify RBMEC from other cell types. Fluorescence of P-gp substrate rhodaminel 23(Rh123) in RBMEC was measured to evaluate the effect of E6 on the function of P-gp. RESULTS: Purified RBMEC was obtained after being sorted from other cell types by flow cytometry. Intracellular fluorescence of Rh123 in RBMEC increased after incubation with E6 3, 10μmol/L or verapamil(VER) 50μmol/L (P<0.01), respectively. After the treatment of E6 10μmol/L, intracellular Rh123 accumulation was increased nearly to 3-fold. E6 and VER have no effect on accumulation of Rh123 in human umbilical vein endothelial cells(HUVEC), which do not express P-gp. CONCLUSION: E6 exhibited a strong inhibitory effect on the activity of P-gp in RBMEC.目的:研究新的钙调素抑制剂E6对纯化的原代培养大鼠脑微血管内皮细胞上P-糖蛋白活性的影响。方法:使用流式细胞仪纯化大鼠脑微血管内皮细胞,通过检测P-糖蛋白底物罗丹明123在大鼠脑微血管内皮细胞中的荧光评估E6对P-糖蛋向功能的影响。结果:使用流式细胞仪能将大鼠脑微血管内皮细胞从其它杂细胞中分离纯化,3,10μmol/L E6和50μmol/L维拉帕米能显著提高大鼠脑微血管内皮细胞内罗丹明123的含量(P<0.01),10μmol/L E6能使罗丹明123的胞内浓度增加接近三倍,而无P-糖蛋白表达的人脐静脉内皮细胞中的罗丹明123浓度则不受影响。结论:E6能显著抑制大鼠脑微血管内皮细胞上P-糖蛋白活性。

关 键 词：E6 P-GLYCOPROTEIN blood-brain barrier 

分 类 号：R96[医药卫生—药理学]