山豆根黄酮类成分对斑马鱼的急性毒性研究

Acute toxicity study of flavonoids from Sophorae Tonkinensis Radix et Rhizoma on zebrafish

目的利用体外细胞毒模型结合模式生物斑马鱼急性毒性模型,筛选和评价山豆根提取物、组分及重要单体成分的细胞毒性及对斑马鱼的急性毒性,以期发现山豆根潜在的毒性成分。方法考察山豆根提取物、组分及主要单体成分对人心肌细胞AC16、人正常肝细胞L-02、人肾小管上皮细胞HK-2和人神经母细胞瘤细胞SH-SY5Y的毒性,筛选山豆根中具有较强细胞毒性的组分及主要单体成分,评价其对模式生物斑马鱼的急性毒性,计算斑马鱼的最大非致死浓度(maximun non-lethal concentration,MNLC)、10%致死浓度(10%lethal concentration,LC10)和半数致死浓度(half lethal concentration,LC50)。给予山豆根酮(1/9MNLC、1/3MNLC、MNLC、LC10)进行干预,观察斑马鱼器官形态学结构的畸形变化、运动情况的异常变化和死亡数量等毒性表型,计算畸形变化率,并考察斑马鱼肝脏组织病理变化。结果山豆根酮对AC16、L-02、HK-2和SH-SY5Y4细胞均具有较强的细胞毒性;在斑马鱼急性毒性模型中,山豆根酮组斑马鱼毒性明显,主要表现为肝毒性、神经毒性、心血管毒性、肾毒性等,其中肝毒性在各剂量下均表现明显,且肝脏组织病理切片显示山豆根酮可诱发斑马鱼肝脏损伤,表明肝脏可能为山豆根酮的主要毒性靶器官。结论山豆根酮为山豆根潜在的毒性成分之一。

Objective To screen and evaluate cytotoxicity of extracts,components and important monomer components of Sophorae Tonkinensis Radix et Rhizoma in cytotoxicity model and acute toxicity on model organism zebrafish acute toxicity model,to discover the potential toxic components of Sophorae Tonkinensis Radix et Rhizoma.Methods Toxicity of extracts,components and important monomer components of Sophorae Tonkinensis Radix et Rhizoma on human cardiomyocyte AC16,human normal hepatocyte L-02,human renal tubular epithelial cell HK-2 and human neuroblastoma cell SH-SY5 Y were investigated.Strong cytotoxic components and main monomer components in Sophorae Tonkinensis Radix et Rhizoma were screened,acute toxicity to the model organism zebrafish were evaluated,maximum non-lethal concentration(MNLC)of zebrafish,10%lethal concentration(LC10)and half lethal concentration(LC50)were calculated.Sophoranone(1/9 MNLC,1/3 MNLC,MNLC,LC10)were used to intervene,abnormal changes in zebrafish organ morphology,abnormal changes in movement,and number of deaths and other toxic phenotypes were observed,abnormal changes rate and pathological changes of liver tissue were evaluated.Results Sophoranone had strong cytotoxicity to AC16 cells,L-02 cells,HK-2 cells and SH-SY5 Y4 cells;Toxic phenotype of zebrafish in sophoranone group was obvious in acute toxicity model of zebrafish,mainly manifested as hepatotoxicity,neurotoxicity,cardiovascular toxicity,nephrotoxicity,among which hepatotoxicity was obvious at each dose,and histopathological slices of liver showed that sophoranone induced zebrafish liver damage,indicating that liver may be the main toxic target organs of Sophorae Tonkinensis Radix et Rhizoma.Conclusion Sophoranone is one of the potential toxic components of Sophorae Tonkinensis Radix et Rhizoma.