【摘要】 目的　研究蛋白酪氨酸激酶和蛋白酪氨酸磷酸酶抑制剂对牛脑血管平滑肌细胞 (CSMC)Ca2 + 池操纵性Ca2 + 内流的影响。方法　采用培养的CSMC ,在生物荧光双波长影像分析系统用Fura 2 /Am荧光探针测定单个细胞内游离Ca2 + 浓度。结果　 (1)蛋白酪氨酸激酶抑制剂 (genistein ,2 5 ,5 ,10 μmol·L-1)能浓度依赖性降低内皮素 1(ET 1,10 -7mol·L-1)刺激引起的CSMCCa2 + 内流 ,抑制率分别为5 6%± 2 .9%、2 5 6%± 3 9%、48 9%± 3 7% ;蛋白酪氨酸磷酸酶抑制剂 (vanadate ,2 ,4,8μmol·L-1)能浓度依赖性升高CPA刺激引起的CSMCCa2 + 内流 ,增加比率分别为8 2 %± 3 9%、18 8%± 4 9%、46 6%± 6 9% ;(2 ) genistein(2 5 ,5 ,10 μmol·L-1)能浓度依赖性降低ATP(10 μmol·L-1)刺激引起的CSMCCa2 + 内流 ,抑制率分别为 6 7%±2 6%、2 4 6%± 6 5 %、5 1 3 %± 6 9% ;vanadate (2 ,4,8μmol·L-1)能浓度依赖性升高ATP刺激引起的CSMCCa2 +内流 ,增加比率分别为 4 8%± 2 0 %、2 8 5 %± 4 6%、49 6%± 3 3 % ;(3 ) genistein (2 5 ,5 ,10 μmol·L-1)能浓度依赖性降低环匹阿尼酸 (Cyclopiazonicacid ,CPA ,10 μmol·L-1)刺激引起的CSMCCa2 + 内流 ,抑制率分别为 6 5 %± 3 0 %、2 2 5 %± 5 2 %、更多还原

【Abstract】 AIM To study the effects of protein tyrosine kinase inhibitor and protein tyrosine phosphatase inhibitor on cultured bovine cerebrovascular smooth muscle cells (CSMC) Ca 2+ store operated Ca 2+ influx. METHODS Cell culture and single intracellular free Ca 2+ concentration was measured in fura 2/Am flueorescence probe by MetaFluor Fluorescence ratio imaging system. RESULTS (1) protein tyrosine kinase inhibitor (genistein,2.5,5,10 μmol·L -1 )decreased Ca 2+ influx significantly induced by endothelin 1(ET 1),ATP,cyclopiazonic acid(CPA) in concentration dependent manner; (2) Protein tyrosine phosphatase inhibitor (vanadate,2,4,8 μmol·L -1 ) increased Ca 2+ influx significantly induced by ET 1,ATP,CPA in concentration dependent manner. CONCLUSION Protein tyrosine kinase inhibitor and protein tyrosine phosphatase inhibitor have effects on Ca 2+ store operated Ca 2+ influx induced by ET 1, ATP, CPA. Protein tyrosine phosphorylation participats in the signal transduction of Ca 2+ store operat Ca 2+ influx in cerebrovascular smooth muscle cells.更多还原