In order to find candidate proteins that are potentially associated with the phenotype in neonatal rat cardiomyocytes treated with phenylephrine(PE), the differential proteins expression pattern after α-adrenergic re...In order to find candidate proteins that are potentially associated with the phenotype in neonatal rat cardiomyocytes treated with phenylephrine(PE), the differential proteins expression pattern after α-adrenergic receptor stimulation in neonatal cardiomyocytes was analyzed by using a series of methods,including two dimensional polyacrylamide gel electrophoresis,PDQuest 2-DE software analysis and peptide mass fingerprint mapping based on MALDI-TOF-MS.The good two-dimensional pattern with a high resolution and reproducibility was obtained.Totally 14 differential protein spots were found. Out of these,11 proteins were preliminarily identified.Desmin,Txndc5,etc.were found to be up-regulated in PE-treated cardiomyocytes,while caspase-11, 4-hydroxyphenylpyruvic acid dioxygenase,etc.were found to be down-regulated.It suggests that the differential expression analysis of proteome may be useful to the further study of the related proteins and the molecular markers of cardiovascular diseases.展开更多
目的：合成α_1-肾上腺素受体特异性激动剂——苯肾上腺素(phenylephrine，PE)荧光标记物，检测其药理学生物活性。方法：用化学合成的方法将绿色荧光染料 BODIPY-FL 和 PE 进行微量缩合反应，薄层色谱法分离和纯化，用薄层色谱和质谱...目的：合成α_1-肾上腺素受体特异性激动剂——苯肾上腺素(phenylephrine，PE)荧光标记物，检测其药理学生物活性。方法：用化学合成的方法将绿色荧光染料 BODIPY-FL 和 PE 进行微量缩合反应，薄层色谱法分离和纯化，用薄层色谱和质谱分析对其纯度和分子结构进行鉴定。通过分子生物学 Western blot 方法检测其药理学生物活性。结果：将 BODIPY-FL 与 PE 在无水、无氧和避光条件下进行羧合反应，经薄层色谱分离、纯化得到具有荧光的新化合物——BODIPY-FL-PE，结构经紫外光谱和质谱分析得到证实。Western blot 实验表明 BODIPY-FL-PE与 PE 同样具有激动α_1-肾上腺素受体，使胞外信号调节激酶(extracellular signal-regulated kinase，ERK)激活的药理作用。结论：经微量缩合反应合成的 BODIPY-FL-PE 具有α_1-肾上腺素受体激动剂药理学特性，可以用于肾上腺素受体行为观测，为进一步研究活细胞中肾上腺素受体的动态行为提供了工具。展开更多
文摘In order to find candidate proteins that are potentially associated with the phenotype in neonatal rat cardiomyocytes treated with phenylephrine(PE), the differential proteins expression pattern after α-adrenergic receptor stimulation in neonatal cardiomyocytes was analyzed by using a series of methods,including two dimensional polyacrylamide gel electrophoresis,PDQuest 2-DE software analysis and peptide mass fingerprint mapping based on MALDI-TOF-MS.The good two-dimensional pattern with a high resolution and reproducibility was obtained.Totally 14 differential protein spots were found. Out of these,11 proteins were preliminarily identified.Desmin,Txndc5,etc.were found to be up-regulated in PE-treated cardiomyocytes,while caspase-11, 4-hydroxyphenylpyruvic acid dioxygenase,etc.were found to be down-regulated.It suggests that the differential expression analysis of proteome may be useful to the further study of the related proteins and the molecular markers of cardiovascular diseases.
文摘目的：合成α_1-肾上腺素受体特异性激动剂——苯肾上腺素(phenylephrine，PE)荧光标记物，检测其药理学生物活性。方法：用化学合成的方法将绿色荧光染料 BODIPY-FL 和 PE 进行微量缩合反应，薄层色谱法分离和纯化，用薄层色谱和质谱分析对其纯度和分子结构进行鉴定。通过分子生物学 Western blot 方法检测其药理学生物活性。结果：将 BODIPY-FL 与 PE 在无水、无氧和避光条件下进行羧合反应，经薄层色谱分离、纯化得到具有荧光的新化合物——BODIPY-FL-PE，结构经紫外光谱和质谱分析得到证实。Western blot 实验表明 BODIPY-FL-PE与 PE 同样具有激动α_1-肾上腺素受体，使胞外信号调节激酶(extracellular signal-regulated kinase，ERK)激活的药理作用。结论：经微量缩合反应合成的 BODIPY-FL-PE 具有α_1-肾上腺素受体激动剂药理学特性，可以用于肾上腺素受体行为观测，为进一步研究活细胞中肾上腺素受体的动态行为提供了工具。
