胡豆莲的急性毒性及其潜在的毒性成分研究

Acute Toxicity Evaluation and Potential Toxic Components of Euchresta Japonica Hook.f.ex Regel

目的检测胡豆莲根、茎、叶各部位对小鼠的急性毒性,建立胡豆莲中8个化学成分的含量测定方法,初步推测其潜在的毒性成分。方法根据预试验测得胡豆莲根、茎和叶95%乙醇提取物引起动物100%死亡的最小给药剂量和0%死亡的最大给药剂量,设置5个剂量组和空白组,对小鼠灌胃进行急性毒性试验,采用改良寇氏法计算LD50和95%可信限;检测最大非致死浓度下小鼠的生化指标、各脏器指数和体质量变化评价其急性毒性;采用HPLC测定其中6个生物碱和2个黄酮的含量;利用Pearson法计算其中8个化学成分含量与LD50之间的相关系数。结果胡豆莲根、茎、叶的小鼠LD50分别为11.969、11.866和15.601 g·kg^(-1)生药量;其根、茎和叶最大非致死浓度下小鼠血液中的谷丙转氨酶、谷草转氨酶和肌酐含量与空白组比有明显升高趋势(P<0.05或P<0.01),胡豆莲根和叶组的血尿素氮无明显变化,胡豆莲茎组小鼠血液中的尿素氮明显升高(P<0.05);8种成分的峰面积与质量浓度呈线性关系,决定系数R2均>0.990,其中,金雀花碱(H3)的含量与LD50相关系数为-0.985。结论胡豆莲根、茎和叶提取物均可引起肝肾功能损伤,具有明显的急性经口毒性,其根和茎的毒性较大,叶的毒性较小,其潜在的毒性成分为金雀花碱。

OBJECTIVE To establish the acute toxicity evaluation of the roots,stems and leaves in mice and quantitative determination method of 8 chemical components in Euchresta japonica,and preliminarily speculate the potential toxic components.METHODS The acute toxicity test was performed by gavage of 95%ethanol extracts of roots,stems and leaves of Euchresta japonica into mice with blank and five dose groups,and the groups were set by the preliminary experiment of the minimum dose cause 100%death and the maximum dose cause 0%death.The modified Kohl's method was used to calculate LD50 and 95%confidence limits;the biochemical indexes,organ indices and body weight changes of mice with maximum tolerance were detected to evaluate the acute toxicity.The contents of 6 alkaloids and 2 flavonoids were determined by HPLC.The correlation coefficient between 8 chemical components with LD50 was calculated by Pearson's method.RESULTS The LD50 of the roots,stems and leaves of Euchresta japonica were 11.969,11.866 and 15.601 g∙kg^(-1),respectively;the concentration of alanine aminotransferase,aspartate aminotransferase and creatinine in the blood of mice in the root,stem and leaf groups were significantly increased under the maximum non-lethal concentration(P<0.05 or P<0.01),while the blood urea nitrogen in the root and leaf groups had no significant change.The blood urea nitrogen level in the stem group was significantly increased(P<0.05).The peak area and mass concentration of the 8 components had linear relationship with the coefficient of determination R2>0.990,of which the correlation coefficient between the content of(-)-cytisine(H3)and the LD50 value was-0.985.CONCLUSION The roots,stems and leaves extracts of Euchresta japonica can cause liver and kidney damage,have showed obvious acute oral toxicity,the roots and stems are more toxic than the leaves,and the potential toxic component is(-)-cytisine.