CAJ | 학술논문

目的：建立实时细胞分析系统( RTCA)心脏毒性研究方法,并应用于药物心脏毒性的早期筛选。方法采用RTCA Cardio系统和新生1~2 d SD大鼠原代培养的心肌细胞建立体外药物心脏毒性早期筛选方法,并用具有抗心律失常药物奎尼丁和利多卡因,通过观察心肌细胞搏动频率、幅度和搏动节律不规则性( BRl)指数的变化,验证其评价药物的心脏毒性的效果。结果原代心肌细胞接种到 RTCA Cardio E-Plate 96孔板24 h后出现心肌细胞搏动,48 h 后心肌细胞搏动规律且稳定,可持续至少3 d。加入奎尼丁后迅速抑制心肌细胞的搏动,使心肌细胞的搏动频率由155±5降到0,6 h 后抑制作用减弱；低浓度奎尼丁(3.1μmol·L－1)使心肌细胞搏动恢复到124±16。24 h奎尼丁浓度小于100.0μmol·L－1时,均能全部恢复细胞搏动。利多卡因加入原代心肌细胞后,心肌细胞的搏动频率、幅度、BRl的变化趋势呈明显的浓度依赖性,浓度越高,对心肌细胞的抑制作用越显著。结论 RTCA Cardio心脏毒性筛选方法可精确地检测出奎尼丁和利多卡因的心脏毒性,提示 RTCA Cardio 心脏毒性筛选方法可用于心脏毒性早期筛选。
목적：건립실시세포분석계통( RTCA)심장독성연구방법,병응용우약물심장독성적조기사선。방법채용RTCA Cardio계통화신생1~2 d SD대서원대배양적심기세포건입체외약물심장독성조기사선방법,병용구유항심률실상약물규니정화리다잡인,통과관찰심기세포박동빈솔、폭도화박동절률불규칙성( BRl)지수적변화,험증기평개약물적심장독성적효과。결과원대심기세포접충도 RTCA Cardio E-Plate 96공판24 h후출현심기세포박동,48 h 후심기세포박동규률차은정,가지속지소3 d。가입규니정후신속억제심기세포적박동,사심기세포적박동빈솔유155±5강도0,6 h 후억제작용감약；저농도규니정(3.1μmol·L－1)사심기세포박동회복도124±16。24 h규니정농도소우100.0μmol·L－1시,균능전부회복세포박동。리다잡인가입원대심기세포후,심기세포적박동빈솔、폭도、BRl적변화추세정명현적농도의뢰성,농도월고,대심기세포적억제작용월현저。결론 RTCA Cardio심장독성사선방법가정학지검측출규니정화리다잡인적심장독성,제시 RTCA Cardio 심장독성사선방법가용우심장독성조기사선。
OBJECTlVE To establish a real-time cell analysis system ( RTCA) for early drug car-diotoxicity evaluation. METHODS An in vitro drug cardiotoxicity evaluation method was established using RTCA Cardio system and primary cultured cardiomyocytes of neonatal rats. The beating rate, am-plitude and beating rhythm irregularity ( BRl ) of cardiomyocytes were observered after antiarrhythmic drugs, such as quinidine and lidocaine were added, to assess the effect of the above method on cardio-toxicity evaluation. RESULTS RTCA Cardo E-Plate 96 was inoculated with primary cultured cardiomyo-cytes that began to beat after 24 h and beat regularly after 48 h. The stable beating was maintained for a minimum of three days. The beating of cardiomyocytes decreased rapidly from 155±5 to 0 after incuba-tion with quinidine. The beating recovered gradually after 6 h. Quinidine at 3.1μmol·L-1 caused the beat-ing rate to return to 124±16. Quinidine allowed the beating rate to return to normal when the concentra-tion was less than 100.0μmol·L-1 . The beating rate, amplitude and BRl of cardiomyocytes changed in a concentration-dependent manner when incubating with lidocaine. The higher the concentration, the more significant the inhibition of lidocaine on cardiomyocytes. CONCLUSlON The cardiotoxicity of quinidine and lidocaine can be detected accurately using RTCA Cardio system, suggesting that this system can be used in early evaluation of drug cardiotoxicity.