蝙蝠葛碱体外肝肾细胞毒性的初步研究

Preliminary Study on Hepatocytes and Nephrocytes Toxicity Induced by Dauricine

目的探讨北豆根中的成分蝙蝠葛碱在体外对人正常肝细胞（L-02）、人胚肾细胞（HEK-293）和人肾小管上皮细胞（HK-2）的影响。方法采用MTT法检测蝙蝠葛碱对肝细胞和肾细胞活力的影响；采用倒置相差显微镜对给药后的细胞形态进行观察；检测给予蝙蝠葛碱后，肝细胞培养上清液中的功能性指标（AST、ALP、LDH）和肾细胞培养上清液中的功能性指标（LDH）的含量。结果MTT法显示，50～100μg·mL^-1的蝙蝠葛碱能显著降低L-02细胞的活力（P〈0．01），25～200μg·mL^-1的蝙蝠葛碱对HEK-293细胞有明显的抑制作用（P〈0．01），12．5～50μg·mL^-1的蝙蝠葛碱能降低HK-2细胞的活力（P〈0．01或P〈0．05）；给予蝙蝠葛碱后，肝肾细胞均不同程度的皱缩、减少、甚至死亡，且随药物浓度的增加细胞呈现此状态的数目增多；100和200μg·mL^-1的蝙蝠葛碱能显著升高肝细胞培养上清中的AST、ALP、LDH含量和肾细胞上清中的LDH含量（P〈0．01）。结论蝙蝠葛碱对肝肾细胞均有毒性作用。

Objective To test the toxic effects of dauricine on human normal hepatic cells（L-02）, human embryonic kidney 293 cells（HEK293） and human renal tubular epithelial cells（HK-2）. Methods MTT assay was used to test the cell viability of dauricine on hepatocytes and nephrocytes. Morphological changes of hepatocytes and nephro- cytes were observed through inverted phase contrast microscope. The contents of LDH in nephrocytes supernatant and AST, ALP, LDH in hepatocytes supernatant were detected. Results The MTT assay showed that 50 - 100μg·mL^-1 dauricine could inhibit L-02 cells viability obviously（P 〈0.01）, 25 - 200μg·mL^-1 dauricine showed significant inhibitory action on HEK-293 cells viability（P 〈0.01）, 12.5 - 50μg·mL^-1 dauricine could inhibit HK-2 cells viability （P〈0.01 or P〈0.05）. Cell morphology showed that dauricine could damage the shapes of hepatocytes and nephrocytes and increase the cells mortality in a dose-dependent manner by microscope. 100 and 200μg·mL^-1 Dauricine could increase the contents of LDH in nephrocytes supernatant and AST, ALP, LDH in hepatocytes supernatant（P 〈0.01）. Conclusion Dauricine may cause hepatocytes and nephrocytes toxicity.