胡豆莲的安全性评估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

关键词:  胡豆莲  急性毒性  生物碱  含量测定  金雀花碱

Key words:  Euchresta japonica  acute toxicity  alkaloid  quantitative determination  (?)-cytisine

摘要:

目的 胡豆莲根、茎、叶各部位对小鼠的安全性评价，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 根据预实验测得胡豆莲根、茎和叶95%乙醇提取物引起动物100%死亡的最小给药剂量（Dm）和0%死亡的最大给药剂量（Dn），设置5个剂量组和空白组，对小鼠灌胃进行急性毒性实验，采用改良寇氏法计算LD50和95%可信限；检测最大非致死浓度（MNLC）下小鼠的生化指标、各脏器指数和体重变化评价其安全性；采用高效液相色谱法测定其中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 胡豆莲的根、茎、叶的小鼠LD50分别为11.969 g?kg?1、11.866 g?kg?1和15.601 g?kg?1生药量；其根、茎和叶MNLC下小鼠血液中的谷丙转氨酶（ALT）、谷草转氨酶（AST）和肌酐（CRE）含量与空白组比有明显升高趋势（**P<0.01），胡豆莲根和叶组的血尿素氮（BUN）无明显变化，胡豆莲茎组小鼠血液中的BUN明显升高（*P<0.05）；8种成分的峰面积(y)与质量浓度(x)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值相关系数为?0.9845。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。

Abstract:

OBJECTIVE The acute toxicity evaluation of the roots, stems and leaves in mice and quantitative determination method of 8 chemical components in Euchresta japonica was established, and the potential toxic components were preliminarily speculated. 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 (Dm) and the maximum dose cause 0% death (Dn). 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 g?kg?1, 11.866 g?kg?1 and 15.601 g?kg?1, respectively; the concentration of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and creatinine (CRE) in the blood of mice in the root, stem and leaf groups were significantly increased under the maximum non-lethal concentration (MNLC) (**P<0.01), while the blood urea nitrogen (BUN) in the root and leaf groups had no significant change. The BUN level in the stem group was significantly increased (*P<0.05). The peak area (y) and mass concentration (x) 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 ?0.9845. CONCLUSION The roots, stems and leaves extracts of Euchresta japonica could 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.