Abstract
Aim:
To investigate the protective effects of arctigenin (ATG), a phenylpropanoid dibenzylbutyrolactone lignan from Arctium lappa L (Compositae), against ER stress in vitro and the underlying mechanisms.
Methods:
A cell-based screening assay for ER stress regulators was established. Cell viability was measured using MTT assay. PCR and Western blotting were used to analyze gene and protein expression. Silencing of the CaMKKβ, LKB1, and AMPKα1 genes was achieved by RNA interference (RNAi). An ATP bioluminescent assay kit was employed to measure the intracellular ATP levels.
Results:
ATG (2.5, 5 and 10 μmol/L) inhibited cell death and unfolded protein response (UPR) in a concentration-dependent manner in cells treated with the ER stress inducer brefeldin A (100 nmol/L). ATG (1, 5 and 10 μmol/L) significantly attenuated protein synthesis in cells through inhibiting mTOR-p70S6K signaling and eEF2 activity, which were partially reversed by silencing AMPKα1 with RNAi. ATG (1-50 μmol/L) reduced intracellular ATP level and activated AMPK through inhibiting complex I-mediated respiration. Pretreatment of cells with the AMPK inhibitor compound C (25 μmol/L) rescued the inhibitory effects of ATG on ER stress. Furthermore, ATG (2.5 and 5 μmol/L) efficiently activated AMPK and reduced the ER stress and cell death induced by palmitate (2 mmol/L) in INS-1 β cells.
Conclusion:
ATG is an effective ER stress alleviator, which protects cells against ER stress through activating AMPK, thus attenuating protein translation and reducing ER load.
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Acknowledgements
This work was supported by the National Science & Technology Major Project “Key New Drug Creation and Manufacturing Program”, China (Number: 2009ZX09301-001 and 2009ZX09103-064); the China Ministry of Science and Technology Research Grant 2008ZX10002-020; and the Shanghai Science and Technology Research Grants 08DZ1971403 and 09JC1416700.
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Supplementary figures are available at website of Acta Pharmacologica Sinica on NPG.
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Supplementary information, Figure S1 (download DOC )
First, ATG protected HepG2 cells from tunicamycin (TM)-induced ER stress. (DOC 1075 kb)
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Gu, Y., Sun, Xx., Ye, Jm. et al. Arctigenin alleviates ER stress via activating AMPK. Acta Pharmacol Sin 33, 941–952 (2012). https://doi.org/10.1038/aps.2012.60
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DOI: https://doi.org/10.1038/aps.2012.60
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