目的:探讨从苦豆子(Sophora alopecuroides)中提取分离的苦豆子总碱、苦参碱、槐定碱对大鼠肾上腺嗜铬细胞瘤PC12细胞的毒性作用及其作用机制。方法:苦豆子总碱、苦参碱、槐定碱分别作用于PC12细胞，采用MTT法分别检测苦豆子总碱、苦参碱、槐定碱对PC12细胞的增殖抑制率;流式细胞术检测PC12细胞的凋亡率;蛋白免疫印迹(Western Blotting)法检测神经元特异性烯醇化酶(neuron-specific enolase，NSE)、过氧化物酶体增殖物激活受体γ辅激活因子-1α(PGC-1α)及核因子E2相关因子(nuclear factor E2 related factor，Nrf)2蛋白表达水平。结果:与正常对照组比较，3种苦豆子生物碱均能抑制PC12细胞增殖，促使细胞发生凋亡，并呈现一定的浓度依赖性(P＜0。05或P＜0。01)，其中苦豆子总碱对PC12细胞的增殖抑制作用和促凋亡作用最为显著(P＜0。01);随着苦豆子总碱、苦参碱、槐定碱作用浓度的增加，PC12细胞NSE的表达显著减弱(P＜0。05或P＜0。01)，而PGC-1α、Nrf 2的表达显著增强(P＜0。05或P＜0。01)。结论:苦豆子总碱、苦参碱、槐定碱对PC12细胞具有一定的细胞毒性，其中苦豆子总碱最为显著，其机制可能是通过细胞氧化应激损伤和线粒体功能障碍造成的。
Inhibitory Effect of Sophora Alopecuroides Alkaloids on PC12 Cells and Its Mechanism
Objective:To explore the toxic effects of total alkaloids of sophora alopecuroides,matrine and sophoridine extracted from Sophora alopecuroides on PC 12 cells and the mechanisms.Methods:The total alkaloids of sophora alopecuroides,matrine and sophoridine were used to intervene PC 12 cells respectively.The inhibition rate of total alkaloids of sophora alopecuroides,matrine and sophoridine on PC 12 cells was detected by MTT method.Flow cytometry was used to detect the apoptosis rate of PC 12 cells.Western blotting was used to detect the NSE,PGC-1α and Nrf 2 protein expression level.Results:Compared with the control group,the three alkaloids of sophora alopecuroides could inhibit the proliferation of PC 12 cells and promote apoptosis in a concentration dependent manner(P＜0.05 or P＜0.01).Among them,the total alkaloids of sophora alopecuroides showed the most significant inhibitory effect on the proliferation and apoptosis of PC 12 cells(P＜0.01).With the increase of the concentration of total alkaloids of sophora alopecuroides,matrine and sophoridine,the expression of NSE in PC 12 cells decreased significantly(P＜0.05 or P＜0.01),while the expression of PGC-1α and Nrf2 was signif-icantly enhanced(P＜0.05 or P＜0.01).Conclusion:Sophora alopecuroides total alkaloids,matrine and sophoridine have certain cytotoxicity to PC 12 cells,and sophora alopecuroides total alkaloid shows the most significant effect.Its mechanism may be caused by oxidative stress injury of cells and mitochondrial dysfunction.
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