Ying Ying, Lan Xiao-Peng, Tian Ye-Ping
Department of Laboratory Examination, Fuzhou General Hospital, Nanjing Military Command, Fuzhou 350025, China.
Yao Xue Xue Bao. 2007 Mar;42(3):269-73.
Binding activity and biologic effect of a novel alpha-melanocyte-stimulating hormone analogue were tested on cells transiently expressing the human melanocortin-1 (MC1), MC3, MC4, and MC5 receptors. The human MC1 and MC5 receptor genes were cloned into the expression vector pcDNA3. 1/ myc-his(-) B. The vectors were transferred to HEK-293 cells by the calcium phosphate method. Stable receptor populations were generated using G418 selection (900 microg x mL(-1)) for subsequent bioassay analysis. K(i) values of the novel alpha-MSH analogue for MC1, MC3, MC4, and MC5 receptors were obtained in competition with [125I]-NDP-MSH for binding studies. The cyclic AMP level was tested by using [3H]-cyclic AMP kit. It is showed that K(i) values of the novel alpha-MSH analogue for MC1, MC3, MC4, and MC5 receptors were (0.159 +/- 0.040), (35.430 +/- 6.743), (19.293 +/- 2.780) and (2.230 +/- 0.670) nmol L(-1), respectively. Its EC50 values for MC1, MC3, MC4, and MC5 receptors were (0.45 +/- 0.07), (7.80 +/- 0.65), (2.55 +/- 0.23) and (0.33 +/- 0.09) nmol L(-1), respectively. In these tests, the novel alpha-MSH analogue is a MC1R and MC5R selective agonist.
在瞬时表达人黑素皮质素-1（MC1）、MC3、MC4和MC5受体的细胞上测试了一种新型α-黑素细胞刺激素类似物的结合活性和生物学效应。将人MC1和MC5受体基因克隆到表达载体pcDNA3.1/myc-his(-)B中。通过磷酸钙法将载体转移到HEK-293细胞中。使用G418选择（900μg×mL(-1)）产生稳定的受体群体，用于后续的生物测定分析。在与[125I]-NDP-MSH竞争结合研究中获得了新型α-MSH类似物对MC1、MC3、MC4和MC5受体的K(i)值。使用[3H]-环磷酸腺苷试剂盒测试环磷酸腺苷水平。结果表明，新型α-MSH类似物对MC1、MC3、MC4和MC5受体的K(i)值分别为(0.159±0.040)、(35.430±6.743)、(19.293±2.780)和(2.230±0.670)nmol L(-1)。其对MC1、MC3、MC4和MC5受体的EC50值分别为(0.45±0.07)、(7.80±0.65)、(2.55±0.23)和(0.33±0.09)nmol L(-1)。在这些测试中，新型α-MSH类似物是一种MC1R和MC5R选择性激动剂。
