乌骨藤提取物对原代新生大鼠心肌细胞的毒性研究

Cytotoxic effect of Marsdeniae Tenacissimae Extracts on primary cultured myocardial cells of neonatal rat

目的探讨乌骨藤提取物(MTE)对原代新生大鼠新生心肌细胞的毒性影响。方法分离和培养SD大鼠心肌细胞,将MTE组分A溶于二甲基亚砜,分别使用20、40、80、160和320μg/ml的MTE和8 000μg/ml的5-氟尿嘧啶作为阳性对照处理原代大鼠心肌细胞24 h,采用MTS法测定不同浓度MTE对体外培养原代大鼠心肌细胞增殖的影响;采用RTCA Cardio系统建立体外心脏毒性筛选模型,对大鼠心肌细胞增殖、搏动进行检测。结果原代心肌细胞加入不同浓度的MTE后,细胞存活受到显著抑制,半数存活浓度(EC50)为207.3μg/ml,随着MTE浓度的升高组分A抑制率增强(均P<0.05)。原代心肌细胞接种到RTCA Cardio E-Plate96孔板上,24h后出现心肌细胞搏动。MTE处理后,细胞指数(CI)值显著下降,240μg/ml MTE组分A处理后心肌细胞搏动迅速受到抑制,搏动频率由126次/min下降至0次/min,且不可恢复;160μg/ml和80μg/ml处理后分别在12h和6h后抑制作用减弱,搏动频率为40次/min和84次/min;40μg/ml基本无影响。结论 MTE组分A对原代新生大鼠心肌细胞具有细胞毒作用,能够显著影响其搏动功能。

Objective To investigate the cytotoxic effect of Marsdeniae tenacissimae extracts（MTE） on primarily cultured myocardial cells of neonatal rats. Methods Primarily cultured myocardial cells of neonatal SD rats were treated with 20, 40, 80, 160 and 320ug/ml MTE, and 8 000ug/ml 5-FU （positive control）, cell proliferation was assessed by MTS method. Cytotoxicity and rhythm changes of myocardial cells were determined by Real-time Cellular Analysis Cardio（RTCA Cardio） system. Results Proliferation of rat myocardial cells was inhibited after MTE treatment in a concentration-dependent manne （P〈0.05）, and the half survival rate （EC~） was 207.3tJg/ml. RTCA Cardio system showed that myocardial cells were beating normally 24h after testing started. And the cell index （CI） was down-regulation significantly with MTE. The cells beating rate decreased frequently from 126 to 0 beatings/rain with the treatment of 2401Jg/ml MTE component A. When the cells were treated with 160ug/ml and 80ug/ml component A, the cell beating was resumed at 12h and 6h with a beating rate of 40 beatings/rain and 84 beatings/min, respectively. Conclusion MTE component A has cytotoxic effect on primary cultured myocardial cells of neonatal rat, and sig- nificantly affects the beating function of myocardium.