CAJ | 학술논문

建立了大鼠纹状体细胞膜色谱前沿分析法，研究5-羟色胺（5-HT）受体5-HT1D与藁本内酯的亲和作用。通过大鼠纹状体组织制备得到色谱固定相，利用酶联免疫吸附剂测定法（ ELISA）分别测定硅胶吸附前后细胞膜悬液中5-HT的量，求得细胞膜固定相上5-HT受体含量为每克硅胶（40.5±2.3）pg。利用细胞膜色谱与液相色谱的离线联用，特异性地识别混合对照品中的舒马普坦和藁本内酯；以不同浓度（24.2~242 nmol／L）的5-HT1D受体激动剂舒马普坦为模型药物，连续通过细胞膜色谱柱，记录舒马普坦的突破曲线，测得舒马普坦与受体作用的平衡解离常数（KD）为389 nmol／L；并将舒马普坦通过不同浓度（37.0~370 nmol／L）的藁本内酯饱和后的细胞膜色谱柱，记录色谱柱饱和前后舒马普坦突破曲线的变化，测得藁本内酯与受体作用的 KD 值为4.21μmol／L。该方法快速、有效，适用于求解存在竞争结合时药物与受体作用的平衡解离常数。
건립료대서문상체세포막색보전연분석법，연구5-간색알（5-HT）수체5-HT1D여고본내지적친화작용。통과대서문상체조직제비득도색보고정상，이용매련면역흡부제측정법（ ELISA）분별측정규효흡부전후세포막현액중5-HT적량，구득세포막고정상상5-HT수체함량위매극규효（40.5±2.3）pg。이용세포막색보여액상색보적리선련용，특이성지식별혼합대조품중적서마보탄화고본내지；이불동농도（24.2~242 nmol／L）적5-HT1D수체격동제서마보탄위모형약물，련속통과세포막색보주，기록서마보탄적돌파곡선，측득서마보탄여수체작용적평형해리상수（KD）위389 nmol／L；병장서마보탄통과불동농도（37.0~370 nmol／L）적고본내지포화후적세포막색보주，기록색보주포화전후서마보탄돌파곡선적변화，측득고본내지여수체작용적 KD 치위4.21μmol／L。해방법쾌속、유효，괄용우구해존재경쟁결합시약물여수체작용적평형해리상수。
A rat striatum cell membrane chromatography( CMC)frontal analysis method was developed for the determination of the equilibrium dissociation constants ( KD) for 5-hydroxytryptamine(5-HT)receptor 5-HT1D-ligustilide interactions. Rat striatum was used for preparation of the cell membrane stationary phase( CMSP). An enzyme-linked immunosorbent assay( ELISA)was applied to determine the 5-HT level of CMSP before and after the adsorp-tion of cell membrane,and the value was(40. 5±2. 3)pg per gram of silica. The CMC-offline-HPLC system was applied to specifically recognize the mixed standard solution of sumatriptan and ligustilide. Sumatriptan,a 5-HT1D agonist of 24. 2 to 242 nmol/L,was pumped through a CMC column continuously,and the breakthrough curves were recorded. For further competitive studies,the mobile phase that contained ligustilide(37. 0-370 nmol/L)was pumped through the column to saturate the binding sites. Afterwards,sumatriptan was propelled towards the column. The breakthrough curves were recorded and compared with those obtained from the column without saturation. KD values obtained using frontal analysis were 389 nmol/L and 4. 21μmol/L for sumatriptan and ligustilide,respectively. The competitive binding study indicated that the CMC method could be a quick and efficient way for determining the KDvalues in drug-receptor interactions.