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Activation of cDCs and iNKT cells contributes to triptolide-induced hepatotoxicity via STING signaling pathway and endoplasmic reticulum stress

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Abstract

Triptolide (TP) exhibits therapeutic potential against multiple diseases. However, its application in clinics is limited by TP-induced hepatoxicity. TP can activate invariant natural killer T (iNKT) cells in the liver, shifting Th1 cytokine bias to Th2 cytokine bias. The damaging role of iNKT cells in TP-induced hepatoxicity has been established, and iNKT cell deficiency can mitigate hepatotoxicity. However, the activation of iNKT cells in vitro by TP requires the presence of antigen-presenting cells. Therefore, we hypothesized that TP could induce dendritic cells (DCs) to activate iNKT cells, thereby leading to hepatotoxicity. The hepatic conventional DCs (cDCs) exhibited immunogenic activities after TP administration, upregulating the expression of CD1d, co-stimulatory molecules, and IL-12. Neutralization with IL-12p40 antibody extenuated TP-induced hepatotoxicity and reduced iNKT cell activation, suggesting that IL-12 could cause liver injury by activating iNKT cells. TP triggered the activation and upregulation of STING signaling pathway and increased endoplasmic reticulum (ER) stress. Downregulation of STING reduced cDC immunogenicity, inhibiting the activation of iNKT cells and hepatic damage. These indicated the regulatory effects of STING pathway on cDCs and iNKT cells, and the important roles it plays in hepatoxicity. ER stress inhibitor, 4-phenylbutyrate (4-PBA), also suppressed iNKT cell activation and liver injury, which might be regulated by the STING signaling pathway. Our results demonstrated the possible mechanisms underlying TP-induced hepatoxicity, where the activation of cDCs and iNKT cells was stimulated by upregulated STING signaling and increased ER stress as a result of TP administration.

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Data availability

The datasets generated or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

ALD:

Alcohol-related liver disease

ALT:

Alanine transaminase

ALP:

Alkaline phosphatase

APAP:

Acetaminophen

α-SMA:

Alpha-smooth muscle actin

AST:

Aspartate transaminase

ATF4:

Activating transcription factor 4

ATF6:

Activating transcription factor 6

APC:

Antigen-presenting cells

BDL:

Bile duct ligation

cDC:

Conventional dendritic cell

CHOP:

C/EBP homologous protein

Con A:

Concanavalin A

DC:

Dendritic cell

ER:

Endoplasmic reticulum

eIF2α:

Eukaryotic translation initiation factor 2 alpha

FasL:

Fas ligand

GRP78:

Glucose-regulated protein 78

H&E:

Hematoxylin and eosin

HFD:

High-fat diet

HSC:

Hepatic stellate cell

iNKT cell:

Invariant natural killer T cell

IRE1α:

Inositol-requiring enzyme 1α

IRF3:

IFN regulatory factor 3

LSEC:

Liver sinusoidal endothelial cell

NAFLD:

Nonalcoholic fatty liver disease

NASH:

Nonalcoholic steatohepatitis

NPC:

Nonparenchymal cell

NK cell:

Natural killer cell

4-PBA:

4-Phenylbutyrate

pDC:

Plasmacytoid DC

PERK:

PKR-like ER kinase

PRR:

Pattern recognition receptor

sXBP1:

Spliced X-box-binding protein

TBK1:

TANK-binding kinase 1

TC:

Total cholesterol

TG:

Triglyceride

TGF-β1:

Transforming growth factor beta-1

TP:

Triptolide

TUNEL:

Terminal dUTP nick-end labeling

UPR:

The unfolded protein response

WT:

Wild type

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Acknowledgements

The authors would like to acknowledge Li Bai (University of Science and Technology of China) for kindly providing iNKT cell-deficient (Jα18−/− mice), DN32.D3 cells, and RBL-CD1d cells. The authors also would like to acknowledge NIH Tetramer Core Facility for generously providing mouse CD1d-PBS-57 tetramer-APC. The authors would appreciate Zhenglin Hao, who works in the Pharmaceutical Animal Experimental Center of China Pharmaceutical University, for his kind help with in vivo experiments.

Funding

The present study was supported by the National Natural Science Foundation of China (No. 81703626, No. 82073948, No. 81973562), and National Innovation and Entrepreneurship Training Program for Undergraduate (No. 202210316040Z).

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Authors and Affiliations

  1. New Drug Screening Center, Jiangsu Center for Pharmacodynamics Research and Evaluation, China Pharmaceutical University, Nanjing, 210009, China

    Xin Chen, Zixun Yu, Cheng Nong, Mingxuan Zhang, Lixin Sun, Luyong Zhang & Xinzhi Wang

  2. Reproductive Medicine Center, Department of Obstetrics and Gynecology, The First Affiliated Hospital of Anhui Medical University, Hefei, 230022, China

    Rufeng Xue

  3. Division of Biosciences, University College London, London, WC1E 6BT, UK

    Yiying Zhang

  4. Center for Drug Research and Development, Guangdong Pharmaceutical University, Guangzhou, 510006, China

    Luyong Zhang

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Contributions

Z.Y., N.C., and X.C. performed the experiments, collected data, and analyzed the data. X.C., X.W., M.Z., and L.S. completed the revision experiments. L.Z. and X.W. contributed to the guidance of experiments and the final manuscript. X.R., L.Z., and X.W. designed the study. X.W. wrote the manuscript and answered the reviewers’ questions. Y.Z. modified the language throughout the text. All the authors reviewed the manuscript.

Corresponding authors

Correspondence to Luyong Zhang or Xinzhi Wang.

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Ethical approval

All procedures involved in this study were approved by the Ethics Committee of China Pharmaceutical University and conformed to the Laboratory Animal Management Committee of Jiangsu Province guidelines (Approval No.: 2110748).

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Xin Chen, Zixun Yu, and Cheng Nong are the co-first author.

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Chen, X., Yu, Z., Nong, C. et al. Activation of cDCs and iNKT cells contributes to triptolide-induced hepatotoxicity via STING signaling pathway and endoplasmic reticulum stress. Cell Biol Toxicol 39, 1753–1772 (2023). https://doi.org/10.1007/s10565-022-09782-6

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