Triptolide (TP) is the main active ingredient of Tripterygium wilfordii Hook.f, which has attracted great interest due to its promising efficacy for autoimmune diseases and tumors. However, severe adverse reactions, especially hepatotoxicity, have restricted its approval in the market. In the present study we explored the role of hepatic natural killer T (NKT) cells in the pathogenesis of TP-induced liver injury in mice. TP (600 μg/kg/day, i.g.) was administered to female mice for 1, 3, or 5 days. We found that administration of TP dose-dependently induced hepatotoxicity, evidenced by the body weight reduction, elevated serum ALT and AST levels, as well as significant histopathological changes in the livers. However, the mice were resistant to the development of TP-induced liver injury when their NKT cells were depleted by injection of anti-NK1.1 mAb (200 μg, i.p.) on days −2 and −1 before TP administration. We further revealed that TP administration activated NKT cells, dominantly releasing Th1 cytokine IFN-γ, recruiting neutrophils and macrophages, and leading to liver damage. After anti-NK1.1 injection, however, the mice mainly secreted Th2 cytokine IL-4 in the livers and exhibited a significantly lower percentage of hepatic infiltrating neutrophils and macrophages upon TP challenge. The activation of NKT cells was associated with the upregulation of Toll-like receptor (TLR) signaling pathway. Collectively, these results demonstrate a novel role of NKT cells contributing to the mechanisms of TP-induced liver injury. More importantly, the regulation of NKT cells may promote effective measures that control drug-induced liver injury.
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Li XJY, Jiang ZZ, Zhang LY. Triptolide: progress on research in pharmacodynamics and toxicology. J Ethnopharmacol. 2014;155:67–79.
Liu Q. Triptolide and its expanding multiple pharmacological functions. Int Immunopharmacol. 2011;11:377–83.
Jiang Z, Huang X, Huang S, Guo H, Wang L, Li X, et al. Sex-related differences of lipid metabolism induced by triptolide: the possible role of the LXRalpha/SREBP-1 signaling pathway. Front Pharmacol. 2016;7:87.
Fu Q, Huang X, Shu B, Xue M, Zhang P, Wang T, et al. Inhibition of mitochondrial respiratory chain is involved in triptolide-induced liver injury. Fitoterapia. 2011;82:1241–8.
Regev A. Drug-induced liver injury and drug development: industry perspective. Semin Liver Dis. 2014;34:227–39.
Van Kaer L, Parekh VV, Wu L. Invariant natural killer T cells: bridging innate and adaptive immunity. Cell Tissue Res. 2011;343:43–55.
Li N, Hua JL. Immune cells in liver regeneration. Oncotarget. 2017;8:3628–39.
Arrenberg P, Maricic I, Kumar V. Sulfatide-mediated activation of type II natural killer T cells prevents hepatic ischemic reperfusion injury in mice. Gastroenterology. 2011;140:646–55.
Deng ZB, Liu YL, Liu CR, Xiang XY, Wang JH, Cheng ZQ, et al. Immature myeloid cells induced by a high-fat diet contribute to liver inflammation. Hepatology. 2009;50:1412–20.
Takeda K, Hayakawa Y, Van Kaer L, Matsuda H, Yagita H, Okumura K. Critical contribution of liver natural killer T cells to a murine model of hepatitis. Proc Natl Acad Sci USA. 2000;97:5498–503.
Cheng L, You Q, Yin H, Holt MP, Ju C. Involvement of natural killer T cells in halothane-induced liver injury in mice. Biochem Pharmacol. 2010;80:255–61.
Guo H, Pan C, Chang B, Wu X, Guo J, Zhou Y, et al. Triptolide improves diabetic nephropathy by regulating Th cell balance and macrophage infiltration in rat models of diabetic nephropathy. Exp Clin Endocr Diab. 2016;124:389–98.
Zhang Y, Ma X. Triptolide inhibits IL-12/IL-23 expression in APCs via CCAAT/enhancer-binding protein alpha. J Immunol. 2010;184:3866–77.
Wang H, Feng D, Park O, Yin S, Gao B. Invariant NKT cell activation induces neutrophil accumulation and hepatitis: opposite regulation by IL-4 and IFN-gamma. Hepatology. 2013;58:1474–85.
Wang X, Jiang Z, Cao W, Yuan Z, Sun L, Zhang L. Th17/Treg imbalance in triptolide-induced liver injury. Fitoterapia. 2014;93:245–51.
Wang Y, Jia L, Wu CY. Triptolide inhibits the differentiation of Th17 cells and suppresses collagen-induced arthritis. ScandJ Immunol. 2008;68:383–90.
Kim S, Lalani S, Parekh VV, Vincent TL, Wu L, Van Kaer L. Impact of bacteria on the phenotype, functions, and therapeutic activities of invariant NKT cells in mice. J Clin Invest. 2008;118:2301–15.
Wilson MT, Johansson C, Olivares-Villagomez D, Singh AK, Stanic AK, Wang CR, et al. The response of natural killer T cells to glycolipid antigens is characterized by surface receptor down-modulation and expansion. Proc Natl Acad Sci USA. 2003;100:10913–8.
Park O, Jeong WI, Wang L, Wang H, Lian ZX, Gershwin ME, et al. Diverse roles of invariant natural killer T cells in liver injury and fibrosis induced by carbon tetrachloride. Hepatology. 2009;49:1683–94.
Watanabe Y, Suzuki O, Haruyama T, Akaike T. Interferon-gamma induces reactive oxygen species and endoplasmic reticulum stress at the hepatic apoptosis. J Cell Biochem. 2003;89:244–53.
Lappas CM, Day YJ, Marshall MA, Engelhard VH, Linden J. Adenosine A2A receptor activation reduces hepatic ischemia reperfusion injury by inhibiting CD1d-dependent NKT cell activation. J Exp Med. 2006;203:2639–48.
Zhang NN, Huang NY, Zhou XK, Luo XL, Liu CY, Zhang Y, et al. Protective effects of IL-4 on Bacillus Calmette-Guerin and lipopolysaccharide induced immunological liver injury in mice. Inflamm Res. 2012;61:17–26.
Kumar V. NKT-cell subsets: promoters and protectors in inflammatory liver disease. J Hepatol. 2013;59:618–20.
Skold M, Xiong X, Illarionov PA, Besra GS, Behar SM. Interplay of cytokines and microbial signals in regulation of CD1d expression and NKT cell activation. J Immunol. 2005;175:3584–93.
O’Neill LAJ, Bowie AG. The family of five: TIR-domain-containing adaptors in Toll-like receptor signalling. Nat Rev Immunol. 2007;7:353–64.
Gong QA, Zhang H, Li JH, Duan LH, Zhong S, Kong XL, et al. High-mobility group box 1 exacerbates concanavalin A-induced hepatic injury in mice. J Mol Med. 2010;88:1289–98.
Kallus SJ, Brandt LJ. The intestinal microbiota and obesity. J Clin Gastroenterol. 2012;46:16–24.
Sadik CD, Kim ND, Luster AD. Neutrophils cascading their way to inflammation. Trends Immunol. 2011;32:452–60.
Margalit M, Ilan Y. Induction of immune tolerance: a role for natural killer T lymphocytes? Liver Int. 2005;25:501–4.
Beldi G, Wu Y, Banz Y, Nowak M, Miller L, Enjyoji K, et al. Natural killer T cell dysfunction in CD39-null mice protects against concanavalin A-induced hepatitis. Hepatology. 2008;48:841–52.
Li ZP, Oben JA, Yang SQ, Lin HZ, Stafford EA, Soloski MJ, et al. Norepinephrine regulates hepatic innate immune system in leptin-deficient mice with nonalcoholic steatohepatitis. Hepatology. 2004;40:434–41.
Ma X, Hua J, Li ZP. Probiotics improve high fat diet-induced hepatic steatosis and insulin resistance by increasing hepatic NKT cells. J Hepatol. 2008;49:821–30.
Kato T, Sato Y, Takahashi S, Kawamura H, Hatakeyama K, Abo T. Involvement of natural killer T cells and granulocytes in the inflammation induced by partial hepatectomy. J Hepatol. 2004;40:285–90.
Hayakawa Y, Takeda K, Yagita H, Kakuta S, Iwakura Y, Van Kaer L, et al. Critical contribution of IFN-gamma and NK cells, but not perforin-mediated cytotoxicity, to anti-metastatic effect of alpha-galactosylceramide. Eur J Immunol. 2001;31:1720–7.
Minagawa M, Oya H, Yamamoto S, Shimizu T, Bannai M, Kawamura H, et al. Intensive expansion of natural killer T cells in the early phase of hepatocyte regeneration after partial hepatectomy in mice and its association with sympathetic nerve activation. Hepatology. 2000;31:907–15.
Downs I, Liu J, Aw TY, Adegboyega PA, Ajuebor MN. The ROS scavenger, NAC, regulates hepatic Valpha14iNKT cells signaling during Fas mAb-dependent fulminant liver failure. PLoS One. 2012;7:e38051.
Kim YH, Kumar A, Chang CH, Pyaram K. Reactive oxygen species regulate the inflammatory function of NKT cells through promyelocytic leukemia zinc finger. J Immunol. 2017;199:3478–87.
He QL, Titov DV, Li J, Tan M, Ye Z, Zhao Y, et al. Covalent modification of a cysteine residue in the XPB subunit of the general transcription factor TFIIH through single epoxide cleavage of the transcription inhibitor triptolide. Angew Chem Int Ed. 2015;54:1859–63.
Wang X, Jiang Z, Xing M, Fu J, Su Y, Sun L, et al. Interleukin-17 mediates triptolide-induced liver injury in mice. Food Chem Toxicol. 2014;71:33–41.
Wang X, Sun L, Zhang L, Jiang Z. Effect of adoptive transfer or depletion of regulatory T cells on triptolide-induced liver injury. Front Pharmacol. 2016;7:99.
Subramanian M, Kini R, Madasu M, Ohta A, Nowak M, Exley M, et al. Extracellular adenosine controls NKT-cell-dependent hepatitis induction. Eur J Immunol. 2014;44:1119–29.
Liu L, Jiang Z, Liu J, Huang X, Wang T, Liu J, et al. Sex differences in subacute toxicity and hepatic microsomal metabolism of triptolide in rats. Toxicology. 2010;271:57–63.
Kawamura T, Takeda K, Kaneda H, Matsumoto H, Hayakawa Y, Raulet DH, et al. NKG2A inhibits invariant NKT cell activation in hepatic injury. J Immunol. 2009;182:250–8.
Shimizu Y, Margenthaler JA, Landeros K, Otomo N, Doherty G, Flye MW. The resistance of P. acnes--primed interferon gamma-deficient mice to low-dose lipopolysaccharide-induced acute liver injury. Hepatology. 2002;35:805–14.
Van Dyken SJ, Locksley RM. Interleukin-4- and interleukin-13-mediated alternatively activated macrophages: roles in homeostasis and disease. Annu Rev Immunol. 2013;31:317–43.
Kwon HJ, Won YS, Park O, Feng DC, Gao B. Opposing effects of prednisolone treatment on T/NKT cell- and hepatotoxin-mediated hepatitis in mice. Hepatology. 2014;59:1094–106.
Matsuoka N, Itoh T, Watarai H, Sekine-Kondo E, Nagata N, Okamoto K, et al. High-mobility group box 1 is involved in the initial events of early loss of transplanted islets in mice. J Clin Invest. 2010;120:735–43.
Reynolds JM, Dong C. Toll-like receptor regulation of effector T lymphocyte function. Trends Immunol. 2013;34:511–9.
Akira S, Takeda K. Toll-like receptor signalling. Nat Rev Immunol. 2004;4:499–511.
Seki E, Park E, Fujimoto J. Toll-like receptor signaling in liver regeneration, fibrosis and carcinogenesis. Hepatol Res. 2011;41:597–610.
The present study was supported by the National Natural Science Foundation of China (No. 81703626, No. 81773995, No. 81773827, No. 81573514, No. 81673684, No. 81673443, No. 81573690, and No. 81320108029), the Fundamental Research Funds for the Central Universities (2632017PY11), the Natural Science Foundation of Jiangsu Province (BK20151439), and grants from the College Students Innovation Project for the R&D of Novel Drugs (J1310032).
X-zW designed the experiments; X-zW, R-fX, and S-yZ performed the experiments; X-zW, Y-tZ, and L-yZ analyzed and discussed the data; X-zW and Z-zJ wrote the paper. All authors contributed to editing of the paper and scientific discussions.
About this article
- drug-induced liver injury
- natural killer T cell
- Th1/Th2 cytokines
- toll-like receptor signaling pathway