Arctigenin alleviates ER stress via activating AMPK.

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 CaMKK?, LKB1, and AMPK?1 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 ?mol/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 ?mol/L) significantly attenuated protein synthesis in cells through inhibiting mTOR-p70S6K signaling and eEF2 activity, which were partially reversed by silencing AMPK?1 with RNAi. ATG (1-50 ?mol/L) reduced intracellular ATP level and activated AMPK through inhibiting complex I-mediated respiration. Pretreatment of cells with the AMPK inhibitor compound C (25 ?mol/L) rescued the inhibitory effects of ATG on ER stress. Furthermore, ATG (2.5 and 5 ?mol/L) efficiently activated AMPK and reduced the ER stress and cell death induced by palmitate (2 mmol/L) in INS-1 ? 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.

SUBMITTER: Gu Y 

PROVIDER: S-EPMC4011149 | biostudies-literature | 2012 Jul

REPOSITORIES: biostudies-literature