丙戊酸钠及其代谢产物的体外肝毒性研究

目的 研究丙戊酸钠及3个代谢产物（2-丙基-4-五烯酸、3-羟基丙戊酸、5-羟基丙戊酸）对体外人正常肝细胞L02增殖活性及对肝细胞损伤相关指标的影响。方法 实验分为对照组和实验组，对照组细胞常规培养，实验组加入丙戊酸钠及3个代谢产物，采用CCK-8法检测细胞增殖活性，PCR法检测CYP1A1、CYP1A2、PCNA、Bax及Bcl-2的mRNA相对含量，Western Blotting法检测蛋白表达，同时检测细胞上清液中谷草转氨酶（AST）、谷丙转氨酶（ALT）、乳酸脱氢酶（LDH）的含量。结果 与对照组相比，随着丙戊酸钠及3个代谢产物浓度和时间的增加，对 L02细胞增殖活性的抑制逐渐增强，CYP1A1、CYP1A2及Bax的mRNA相对含量和蛋白表达量升高，PCNA及Bcl-2的 mRNA相对含量和蛋白表达量均有下降，AST、ALT、LDH含量升高。结论 丙戊酸钠及3个代谢产物与肝毒性有关。

An in vitro study of hepatotoxicity induced by valproic acid and its metabolites

Objective To confirm the hepatotoxicity of valproic acid (VPA) and its metabolites(2-propyl-4-pentenoic acid, 3-hydroxy valproic acid, 5-hydroxy valproic acid) on human liver cells.Methods Cells were divided into control group and VPA-treated group. The control group was conventionally cultured while the VPA-treated group was treated with valproic acid and its metabolites. The rate of cell proliferation was assayed by CCK 8 protocol. The mRNA levels of CYP1A1, CYP1A2, PCNA, Bax and Bcl-2 were measured by real time PCR. The correlated protein levels were measured by Western Blotting. The activity of LDH, AST and ALT were also detected.Results Compared to the control group, with the increases of concentrations and reaction time of VPA and its metabolites, the proliferation rate of L02-cell was reduced, the mRNA and protein levels of CYP1A1, CYP1A2, and Bax was increased, the mRNA and protein level of PCNA and Bcl-2 was decreased, AST, ALT, and LDH were also elevated in the treated group.Conclusion Valproic acid and its metabolites were positively related to hepatotoxicity.