The Resveratrol Attenuates Ethanol-Induced Hepatocyte Apoptosis Via Inhibiting ER-Related Caspase-12 Activation and PDE Activity In Vitro
Article first published online: 13 NOV 2013
Copyright © 2013 by the Research Society on Alcoholism
Alcoholism: Clinical and Experimental Research
Volume 38, Issue 3, pages 683–693, March 2014
How to Cite
Liu, L., Fan, Z., Tang, Y. and Ke, Z. (2014), The Resveratrol Attenuates Ethanol-Induced Hepatocyte Apoptosis Via Inhibiting ER-Related Caspase-12 Activation and PDE Activity In Vitro. Alcoholism: Clinical and Experimental Research, 38: 683–693. doi: 10.1111/acer.12311
- Issue published online: 15 MAR 2014
- Article first published online: 13 NOV 2013
- Manuscript Accepted: 16 SEP 2013
- Manuscript Received: 15 JAN 2013
- National Natural Science Foundation of China. Grant Numbers: 30870812, 30570580
- Endoplasmic Reticulum Stress;
- Caspase-12 Activation;
Endoplasmic reticulum (ER) stress plays a key role in cell apoptosis pathways. Caspase-12, a proapoptotic gene induced by ER stress, is also the key molecule in ER-related apoptosis. The purpose of this study is to evaluate the protective activity and possible mechanism of resveratrol (ResV) against ethanol (EtOH)-induced apoptosis in human hepatocyte Chang cell line.
The human hepatocyte Chang cell line was used to test the hypothesis that ResV may alleviate the liver cell apoptosis induced by EtOH. ER stress-inducible proteins and silent mating type information regulation 2 homolog 1 (SIRT1) were assayed by Western blot. Cell viability was studied by MTT assay and apoptosis was measured by Annexin-V and propidium iodide assay. Caspase-12 activation was examined by immunofluorescence staining. Alcohol dehydrogenase-2 (ADH-2) and aldehyde dehydrogenase-2 (ALDH-2) were measured by polymerase chain reaction amplified product length polymorphism. Phosphodiesterase (PDE) activity was assayed in cell lysates using a cyclic nucleotide PDE assay.
EtOH exposure significantly increased the expression of ER stress markers and activated signaling pathways associated with ER stress. These include GRP78, p-IRE1α, p-eIF2α, p-PERK, ATF4 as well as cleaved caspase-3/12, CHOP/GADD153, and Bax in human hepatocyte Chang cell line. The expression of these proteins were significantly down-regulated by ResV (10 μM) in a SIRT1-dependent manner. ResV can inhibit EtOH-, tunicamycin-, thapsigargin-induced caspase-12 activation. ADH-2 and ALDH-2 activities are lower in this cell line. PDE activity increased by EtOH was inhibited by ResV (10 μM).
The results indicate that (i) EtOH-induced activation of caspase-12 could be one of the underlying mechanisms of hepatocyte apoptosis; (ii) EtOH-induced cell apoptosis was alleviated via ResV (10 μM) by inhibiting ER stress and caspase-12 activation in a SIRT1-dependent manner; and (iii) SIRT1 activated indirectly by ResV (10 μM) attenuates EtOH-induced hepatocyte apoptosis partly through inhibiting PDE activity.