Erk1/2, CDK8, Src and Ck1e Mediate Evodia rutaecarpa Induced Hepatotoxicity in Mice


Evodia rutaecarpa (E.R.) is a commonly used Chinese herbal medicine. However, it exerts certain hepatotoxicity and the underlying molecular mechanism has not been clarified. In this study, we investigated the molecular mechanism involved in hepatotoxicity induced by E.R. Mice were treated with E.R. water- and ethanol-extract at dosage equivalent to 16.67 g crude-drug/kg body weight by intragastric administration once a day on 30 consecutive days. The effect of E.R. extract on liver, manifested by histopathologic effects, liver index, and blood biochemical indexes were tested. In addition, interleukin (IL)-1β, IL-6, IL-8, and tumor necrosis factor (TNF)-α in liver tissue were measured. The signaling transduction molecules were determined by antibody microarray assay, and verified by western blot. E.R. extract, either water- or ethanol-extract, can induce liver dysfunction. Signaling molecules, Erk1/2, Src, CDK8 and CK1e, were involved in this process. E.R. extract can induce Ck1e expression and phosphorylation of Erk1/2 and CDK8, and inhibit Src phosphorylation. Inflammatory cytokines in liver tissue, IL-1β, IL-6, IL-8, and TNF-α were markedly increased upon the treatment of E.R. extract. In conclusion, E.R.-induced hepatotoxicity was due to the expression of inflammatory cytokine, which was mediated through Erk1/2, Src, CDK8 and CK1e.

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Liao, W. , Li, B. , Li, L. , Yan, L. , Wang, Z. , An, X. and Zhao, J. (2015) Erk1/2, CDK8, Src and Ck1e Mediate Evodia rutaecarpa Induced Hepatotoxicity in Mice. Chinese Medicine, 6, 97-108. doi: 10.4236/cm.2015.62011.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Kobayashi, Y. (2003) The Nociceptive and Anti-Nociceptive Effects of Evodiamine from Fruits of Evodia rutaecarpa in Mice. Planta Medica, 69, 425-428.
[2] Moon, T.C., Murakami, M., Kudo, I., et al. (1999) A New Class of COX-2 Inhibitor, Rutaecarpine from Evodia rutaecarpa. Inflammation Research, 48, 621-625.
[3] Peng, J. and Li, Y.J. (2010) The Vanilloid Receptor TRPV1: Role in Cardiovascular and Gastrointestinal Protection. European Journal of Pharmacology, 627, 1-7.
[4] Wu, C.L., Hung, C.R., Chang, F.Y., et al. (2002) Effects of Evodiamine on Gastrointestinal Motility in Male Rats. European Journal of Pharmacology, 457, 169-176.
[5] Huang, X., Li, W. and Yang, X.W. (2012) New Cytotoxic Quinolone Alkaloids from Fruits of Evodia rutaecarpa. Fitoterapia, 83, 709-714.
[6] Kima, D., Lee, Y.H., Park, S.H., et al. (2014) Subchronic Oral Toxicity of Evodia Fruit Powder in Rats. Journal of Ethnopharmacology, 151, 1072-1078.
[7] Hall, D., Ptacek, J. and Snyder, M. (2007) Protein Microarray Technology. Mechanisms of Ageing and Development, 128, 161-167.
[8] Morris, M.K., Chi, A., Melas, I.N., et al. (2013) Phosphoproteomics in Drug Discovery. Drug Discovery Today, 19, 425-432.
[9] Breitling, F., Nesterov, A., Stadler, V., et al. (2009) High-Density Peptide Arrays. Molecular BioSystems, 5, 224-234.
[10] Pierobon, M., VanMeter, A., Moroni, N., et al. (2012) Reverse-Phase Protein Microarrays. Methods in Molecular Biology, 823, 215-235.
[11] Wildt, R.M., Mundy, C.R., Gorick, B.D. and Tomlinson, I.M. (2000) Antibody Arrays for High-Throughput Screening of Antibody-Antigen Interactions. Nature Biotechnology, 18, 989-994.
[12] Cho, Y.E., Singh, T.S., Lee, H.C., Moon, P.G., Lee, J.E., Lee, M.H., et al. (2012) In-Depth Identification of Pathways Related to Cisplatin-Induced Hepatotoxicity through an Integrative Method Based on an Informatics-Assisted Label-Free Protein Quantitation and Microarray Gene Expression Approach. Molecular & Cellular Proteomics, 11, Article ID: M111.010884.
[13] Cho, Y.E., Moon, P.G., Lee, J.E., Singh, T.S.K., Kang, W., Lee, H.-C., et al. (2013) Integrative Analysis of Proteomic and Transcriptomic Data for Iden-tification of Pathways Related to Simvastatin-Induced Hepatotoxicity. Proteomics, 13, 1257-1275.
[14] Hu, Z.Y., Lausted, C., Yoo, H., Yan, X.W., Brightman, A., Chen, J.K., et al. (2014) Quantitative Liver-Specific Protein Fingerprint in Blood: A Signature for Hepatotoxicity. Theranostics, 4, 215-228.
[15] Bancerek, J., Poss, Z.C., Steinparzer, I., Sedlyarov, V., Pfaffenwimmer, T., Mikulic, I., et al. (2013) CDK8 Kinase Phosphorylates Transcription Factor STAT1 to Selectively Regulate the Interferon Response. Immunity, 38, 250-262.
[16] Gustafson, B. and Smith, U. (2006) Cytokines Promote Wnt Signaling and Inflammation and Impair the Normal Differentiation and Lipid Accumulation in 3T3-L1 Preadipocytes. The Journal of Biological Chemistry, 281, 9507-9516.
[17] Hayashi, R., Yamashita, N., Matsui, S., Fujita, T., Araya, J., Sassa, K., et al. (2000) Bradykinin Stimulates IL-6 and IL-8 Production by Human Lung Fibroblasts through ERK- and p38 MAPK-Dependent Mechanisms. European Respiratory Journal, 16, 452-458.
[18] Jiang, J.L. and Hu, C.P. (2009) Evodiamine: A Novel Anti-Cancer Alkaloid from Evodia rutaecarpa. Molecules, 14, 1852-1859.
[19] Kobayashi, Y., Nakano, Y., Kizaki, M., Hoshikuma, K., Yokoo, Y. and Kamiya, T. (2001) Capsaicin-Like Anti-Obese Activities of Evodiamine from Fruits of Evodia rutaecarpa, a Vanilloid Receptor Agonist. Planta Medica, 67, 628-633.
[20] Wang, T., Wang, Y.X., Kontani, Y., Kobayashi, Y., Sato, Y., Mori, N. and Yamashita, H. (2008) Evodiamine Improves Diet-Induced Obesity in a Uncoupling Protein-1-Independent Manner: Involvement of Antiadipogenic Mechanism and Extracellularly Regulated Kinase/Mitogen-Activated Protein Kinase Signaling. Endocrinology, 149, 358-366.
[21] Zhang, T., Qu, S., Shi, Q., He, D.L. and Jin, X.B. (2014) Evodiamine Induces Apoptosis and Enhances TRAIL-Induced Apoptosis in Human Bladder Cancer Cells through mTOR/S6K1-Mediated Downregulation of Mcl-1. International Journal of Molecular Sciences, 15, 3154-3171.
[22] Wang, T., Wang, Y.X. and Yamashita, H. (2009) Evodiamine Inhibits Adipogenesis via the EGFR-PKCα-ERK Signaling Pathway. FEBS Letters, 583, 3655-3659.
[23] Liao, J.F., Chiou, W.F., Shen, Y.C., Wang, G.-J. and Chen, C.-F. (2011) Anti-Inflammatory and Anti-Infectious Effects of Evodia rutaecarpa (Wuzhuyu) and Its Major Bioactive Components. Chinese Medicine, 6, 6-15.
[24] Kong, Y.C., Hu, S.Y., Lau, F.K., Che, C.T., Yueng, H.W., Cheung, S. and Hwang, J.C.C. (1976) Potential Anti-Fertility Plants from Chinese Medicine. The American Journal of Chinese Medicine, 4, 105-128.
[25] King, C.L., Kong, Y.C., Wong, N.S., Yeung, H.W., Fong, H.H.S. and Sankawa, U. (1980) Uterotonic Effect of Evodia rutaecarpa Alkaloids. Journal of Natural Products, 43, 577-582.
[26] Sheu, J.R., Hung, W.C., Wu, C.H., Lee, Y.M. and Yen, M.H. (2000) Antithrombotic Effect of Rutaecarpine, an Alkaloid Isolated from Evodia rutaecarpa, on Platelet Plug Formation in in Vivo Experiments. British Journal of Haematology, 110, 110-115.
[27] Zhu, L.I., Yang, D.X., Liu, X., Jia, F.-L., Ruan, M. and Zhang, B.-X. (2013) Hepatic Toxicity Study on the Fruit of Evodia rutaecarpa (Juss.) Benth. Lishizhen Medicine and Material Medical Research, 24, 1810-1813.
[28] Chou, C.C., Pan, S.L., Teng, C.M. and Guh, J.H. (2003) Pharmacological Evaluation of Several Major Ingredients of Chinese Herbal Medicines in Human Hepatoma Hep3B Cells. European Journal of Pharmaceutical Sciences, 19, 403-412.
[29] LoPiccolo, J., Blumenthal, G., Bernstein, W. and Dennis, P. (2008) Targeting the PI3K/Akt/mTOR Pathway: Effective Combinations and Clinical Considerations. Drug Resistance Updates, 11, 32-50.
[30] Yuana, L.Y. and Kaplowitz, N. (2009) Glutathione in Liver Diseases and Hepatotoxicity. Molecular Aspects of Medicine, 30, 29-41.
[31] Yoon, J.Y., Jeong, H.Y., Kim, S.H., Kim, H.G., Nam, G., Kim, J.P., et al. (2013) Methanol Extract of Evodia lepta Displays Syk/Src-Targeted Anti-Inflammatory Activity. Journal of Ethnopharmacology, 148, 999-1007.
[32] Cubero, F.J. and Nieto, N. (2012) Arachidonic acid Stimulates TNFα Production in Kupffer Cells via a Reactive Oxygen Species-pERK1/2-Egr1-Dependent Mechanism. AJP: Gastrointestinal and Liver Physiology, 303, G228-G239.
[33] Shi, H.L., Wu, X.J., Liu, Y. and Xie, J.Q. (2013) Berberine Counteracts Enhanced IL-8 Expression of AGS Cells Induced by Evodiamine. Life Sciences, 93, 830-839.
[34] Du, J., Wang, X.F., Zhou, Q.M., Zhang, T.L., Lu, Y.Y., Zhang, H. and Su, S.B. (2013) Evodiamine Induces Apoptosis and Inhibits Metastasis in MDA-MB-231 Human Breast Cancer Cells in Vitro and in Vivo. Oncology Reports, 30, 685-694.
[35] Behari, J. (2010) The Wnt/β-Catenin Signaling Pathway in Liver Biology and Disease. Expert Review of Gastroenterology & Hepatology, 4, 745-756.
[36] Lade, A.G. and Monga, S.P. (2011) Beta-Catenin Signaling in Hepatic Development and Progenitors: Which Way Does the WNT Blow? Developmental Dynamics, 240, 486-500.
[37] Firestein, R., Bass, A.J., Kim, S.Y., Dunn, I.F., Silver, S.J., Guney, I., et al. (2008) CDK8 Is a Colorectal Cancer Oncogene That Regulates β-Catenin Activity. Nature, 455, 547-551.
[38] Firestein, R. and Hahn, W.C. (2009) Revving the Throttle on an Oncogene: CDK8 Takes the Driver Seat. Cancer Research, 69, 7899-7901.
[39] Morris, E., Ji, J.Y., Yang, F., Di Stefano, L., Herr, A., Moon, N.S., et al. (2008) E2F1 Represses β-Catenin Transcription and Is Antagonized by both pRB and CDK8. Nature, 455, 552-556.
[40] Kim, S.Y., Dunn, I.F., Firestein, R., Gupta, P., Wardwell, L., Repich, K., et al. (2010) CK1ε Is Required for Breast Cancers Dependent on β-Catenin Activity. PLoS ONE, 5, e8979.
[41] Bibiana, M., Rahaima, R.J., Choi, J.Y., Noguchi, Y., Schürer, S., Chen, W.M., et al. (2013) Development of Highly Selective Casein Kinase 1δ/1ε (CK1δ/ε) Inhibitors with Potent Antiproliferative Properties. Bioorganic & Medicinal Chemistry Letters, 23, 4374-4380.
[42] Condello, S., Cao, L. and Matei, D. (2013) Tissue Transglutaminase Regulates β-Catenin Signaling through a C-Src-Dependent Mechanism. The FASEB Journal, 27, 3100-3112.
[43] Hinck, L., Näthke, I.S., Papkoff, J. and Nelson, W.J. (1994) Beta-Catenin: A Common Target for the Regulation of Cell Adhesion by Wnt-1 and Src Signaling Pathways. Trends in Biochemical Sciences, 19, 538-542.
[44] Bentli, R., Ciftci, O., Cetin, A., Unlu, M., Basak, N. and Cay, M. (2013) Oral Administration of Hesperidin, a Citrus Flavonone, in Rats Counteracts the Oxidative Stress, the Inflammatory Cytokine Production, and the Hepatotoxicity Induced by the Ingestion of 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD). European Cytokine Network, 24, 91-96.
[45] Jaeschke, H., Williams, C.D., Ramachandran, A. and Bajt, M.L. (2012) Acetaminophen Hepatotoxicity and Repair: The Role of Sterile Inflammation and Innate Immunity. Liver International, 32, 8-20.
[46] Saab, L., Peluso, J., Muller, C.D. and Ubeaud-Sequier, G. (2013) Implication of Hepatic Transporters (MDR1 and MRP2) in Inflammation-Associated Idiosyncratic Drug-Induced Hepatotoxicity Investigated by Microvolume Cytometry. Cytometry Part A, 83, 403-408.
[47] Chang, C.P., Chang, J.Y., Wang, F.-Y., Tseng, J. and Chang, J.-G. (1995) The Effect of E.R. Extract on Cytokine Secretion by Human Mononuclear Cells in Vitro. The American Journal of Chinese Medicine, 23, 173-180.
[48] Chang, J.Y., Yang, T.Y., Chang, C.P. and Chang, J.G. (1996) The Effect of “Chi-Han (Hot Nature)” Chinese Herbs on the Secretion of IL-1 Beta and TNF-Alpha by Mononuclear Cells. Journal of Medical Sciences, 12, 8-24.

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