Abstract: Radix Sophorae tonkinensis (RST) is a widely used herb in Traditional Chinese Medicine (TCM) for treating infectious and inflammatory diseases. However, the toxicity data for RST are limited. The aim of this work is to assess and compare the toxicity of the whole RST extract and its five active fractions using the zebrafish model. Five active fractions of RST were prepared using five different types of solvents, which included dealkalized water, ethanol, n-butyl ethanol, dichloromethane, and diethyl ether. The chemical profiles of the active fractions were determined by high-performance liquid chromatography (HPLC), and the toxicity observed in the zebrafish model was confirmed using mouse models. In the zebrafish model, cardiovascular toxicity was observed for the fraction extracted using diethyl ether, and hepatotoxicity was observed for the whole RST extract and the fractions extracted using water and ethanol, whereas both cardiovascular and hepatic toxicities were observed for the fractions extracted using n-butyl ethanol and dichloromethane. The hepatotoxicity of the fractions extracted using n-butyl ethanol and dichloromethane was also observed in mice. Our findings provide the toxicity data for RST and its five active fractions through modeling in a zebrafish, and indicate that the different fractions may each have a different toxicity, which is helpful for the optimal use of RST in clinical practice.

使用斑马鱼模型比较山豆根不同有效部位的毒性

[1]Chai, N.L., Fu, Q., Shi, H., et al., 2012. Oxymatrine liposome attenuates hepatic fibrosis via targeting hepatic stellate cells. World J. Gastroenterol., 18(31):4199-4206.

[2]Cho, C.H., Chuang, C.Y., Chen, C.F., 1986. Study of the antipyretic activity of matrine. A lupin alkaloid isolated from Sophora subprostrata. Planta Med., 52(5):343-345.

[3]Chui, C.H., Lau, F.Y., Tang, J.C., et al., 2005. Activities of fresh juice of Scutellaria barbata and warmed water extract of Radix Sophorae Tonkinensis on anti-proliferation and apoptosis of human cancer cell lines. Int. J. Mol. Med., 16(2):337-341.

[4]CPC (Chinese Pharmacopoeia Commission), 2015. Pharmacopoeia of the People’s Republic of China (Part I). China Medical Science Press, Beijing, China, p.25-26 (in Chinese).

[5]Ding, P.L., Chen, D.F., 2006. Isoprenylated flavonoids from the roots and rhizomes of Sophora tonkinensis. Helv. Chim. Acta, 89(1):103-110.

[6]Ding, P.L., Huang, H., Zhou, P., et al., 2006. Quinolizidine alkaloids with anti-HBV activity from Sophora tonkinensis. Planta Med., 72(9):854-856.

[7]He, C.M., Cheng, Z.H., Chen, D.F., 2013. Qualitative and quantitative analysis of flavonoids in Sophora tonkinensis by LC/MS and HPLC. Chin. J. Nat. Med., 11(6):690-698.

[8]He, J.H., Guo, S.Y., Zhu, F., et al., 2013. A zebrafish phenotypic assay for assessing drug-induced hepatotoxicity. J. Pharmacol. Toxicol. Methods, 67(1):25-32.

[9]Hill, A., 2011. Hepatotoxicity testing in larval zebrafish. In: McGrath, P. (Ed.), Zebrafish: Methods for Assessing Drug Safety and Toxicity. John Wiley & Sons, Inc., Hoboken, NJ, USA, p.89-102.

[10]Hill, A.J., Teraoka, H., Heideman, W., et al., 2005. Zebrafish as a model vertebrate for investigating chemical toxicity. Toxicol. Sci., 86(1):6-19.

[11]Hill, A., Ball, J., Jones, M., et al., 2008. Implementation of zebrafish toxicity testing between in vitro and in vivo models to advance candidate selection. The 29th Annual Meeting of the American College of Toxicology, Tucson, AZ, USA, p.9-12.

[12]Hill, A., Mesens, N., Steemans, M., et al., 2012. Comparisons between in vitro whole cell imaging and in vivo zebrafish-based approaches for identifying potential human hepatotoxicants earlier in pharmaceutical development. Drug Metab. Rev., 44(1):127-140.

[13]Jones, K.S., Alimov, A.P., Rilo, H.L., et al., 2008. A high throughput live transparent animal bioassay to identify non-toxic small molecules or genes that regulate vertebrate fat metabolism for obesity drug development. Nutr. Metab. (Lond.), 5:23.

[14]Kimmel, C.B., Ballard, W.W., Kimmel, S.R., et al., 1995. Stages of embryonic development of the zebrafish. Dev. Dynam., 203(3):253-310.

[15]Lee, J.W., Lee, J.H., Lee, C., et al., 2015. Inhibitory constituents of Sophora tonkinensis on nitric oxide production in RAW 264.7 macrophages. Bioorg. Med. Chem. Lett., 25(4):960-962.

[16]Li, S.J., Yang, J., Qian, X.L., et al., 2011. Experimental study on chronic toxicity in rats caused by water extract components of radix et Rhizoma Sophorae tonkinensise. Chin. J. Pharmacovigil., 8(2):89-92 (in Chinese).

[17]Li, X.N., Sha, N., Yan, H.X., et al., 2008a. Isoprenylated flavonoids from the roots of Sophora tonkinensis. Phytochem. Lett., 1(3):163-167.

[18]Li, X.N., Lu, Z.Q., Chen, G.T., et al., 2008b. NMR spectral assignments of isoprenylated flavanones from Sophora tonkinensis. Magn. Reson. Chem., 46(9):898-902.

[19]Li, X., Luan, Y., Li, X., 2012. Study on anti-inflammatory efficacy accompanied by side effects of different components of Sophorae tonkinensis radix et Rhizoma. China J. Chin. Mater. Med., 37(15):2232-2237 (in Chinese).

[20]Liu, X.S., Jiang, J., Jiao, X.Y., et al., 2006. Matrine-induced apoptosis in leukemia U937 cells: involvement of caspases activation and MAPK-independent pathways. Planta Med., 72(6):501-506.

[21]Long, Y., Lin, X.T., Zeng, K.L., et al., 2004. Efficacy of intramuscular matrine in the treatment of chronic hepatitis B. Hepatobiliary Pancreat. Dis. Int., 3(1):69-72.

[22]McGrath, P., Li, C.Q., 2008. Zebrafish: a predictive model for assessing drug-induced toxicity. Drug Discov. Today, 13(9-10):394-401.

[23]Pan, Q.M., Li, Y.H., Hua, J., et al., 2015. Antiviral matrine-type alkaloids from the rhizomes of Sophora tonkinensis. J. Nat. Prod., 78(7):1683-1688.

[24]Qian, H., Zhao, B.T., Chan, B., et al., 2015. Relationship between the content of polysaccharides, flavonoids and polyphenols from the sporocarp of Phellinus linteus and the antioxidant activity. Sci. Technol. Food Ind., 36(12):104-108 (in Chinese).

[25]Rekha, R.D., Amali, A.A., Her, G.M., et al., 2008. Thioacetamide accelerates steatohepatitis, cirrhosis and HCC by expressing HCV core protein in transgenic zebrafish Danio rerio. Toxicology, 243(1-2):11-22.

[26]Selderslaghs, I.W.T., Blust, R., Witters, H.E., 2012. Feasibility study of the zebrafish assay as an alternative method to screen for developmental toxicity and embryotoxicity using a training set of 27 compounds. Reprod. Toxicol., 33(2):142-154.

[27]Shen, B., Liu, H.C., Ou, W.B., et al., 2015. Toxicity induced by Basic Violet 14, Direct Red 28 and Acid Red 26 in zebrafish larvae. J. Appl. Toxicol., 35(12):1473-1480.

[28]Sun, R., Yang, Q., Zhao, Y., 2010. Comparative study on acute toxicity of different components of radix et Rhizome Sophorae Tonkinensis in mice. Chin. J. Pharmacovigil., 7(5):257-262 (in Chinese).

[29]Tang, L., Dong, L.N., Peng, X.J., et al., 2013. Pharmacokinetic characterization of oxymatrine and matrine in rats after oral administration of radix Sophorae tonkinensis extract and oxymatrine by sensitive and robust UPLC-MS/MS method. J. Pharm. Biomed. Anal., 83:179-185.

[30]Wang, S.H., Li, T.F., Ran, B.D., et al., 2004. Analysis on contents of organic acids and volatile components in tabacco leaves of Yunnan Province. Chin. Tob. Sci., (2):35-37 (in Chinese).

[31]Westerfield, M., 1995. The Zebrafish Book: A Guide for the Laboratory Use of Zebrafish (Danio rerio). University of Oregon Press, Eugene, OR, USA.

[32]Xiao, P., Kubo, H., Komiya, H., et al., 1999. (−)-14β-Acetoxymatrine and (+)-14α-acetoxymatrine, two new matrine-type lupin alkaloids from the leaves of Sophora tonkinensis. Chem. Pharm. Bull. (Tokyo), 47(3):448-450.

[33]Yang, Q., Zheng, L.N., Xie, Y.Z., et al., 2010. Study on the “dosage-time-toxicity” relationship of hepatotoxicity induced by different components of radix et Rhizoma Sophorae Tonkinensis in mice. Chin. J. Pharmacovigil., 7(7):385-389 (in Chinese).

[34]Yoo, H., Chae, H.S., Kim, Y.M., et al., 2014. Flavonoids and arylbenzofurans from the rhizomes and roots of Sophora tonkinensis with IL-6 production inhibitory activity. Bioorg. Med. Chem. Lett., 24(24):5644-5647.

[35]Zhang, C., Willett, C., Fremgen, T., 2003. Zebrafish: an animal model for toxicological studies. Curr. Protoc. Toxicol., 17: 1.7.1-1.7.18.

[36]Zhang, H.Y., Ding, T.H., 2013. Survey of clinical application and toxic reaction of shandougen. Western J. Tradit. Chin. Med., 26(3):121-124 (in Chinese).

[37]Zhou, H.J., Ma, H.L., Guo, D.Z., et al., 2015. Physicochemical properties and antioxidant activity of intracellular polysaccharides from Phellinus igniarius precipitated by different ethanol concentrations. Food Sci., 36(19):34-38 (in Chinese).

[38]Zhou, J., Guo, S.Y., Zhang, Y., et al., 2014. Human prokinetic drugs promote gastrointestinal motility in zebrafish. Neurogastroenterol. Motil., 26(4):589-595.

[39]Zhu, J.J., Xu, Y.Q., He, J.H., et al., 2014. Human cardiotoxic drugs delivered by soaking and microinjection induce cardiovascular toxicity in zebrafish. J. Appl. Toxicol., 34(2):139-148.

[40]Zhu, X.Y., Liu, H.C., Guo, S.Y., et al., 2016. A zebrafish thrombosis model for assessing antithrombotic drugs. Zebrafish, 13(4):335-344.

[41]Zon, L.I., Peterson, R.T., 2005. In vivo drug discovery in the zebrafish. Nat. Rev. Drug Discov., 4(1):35-44.