Involvement of AMPK on the synergistic upregulation of phase II detoxifying enzyme activities by 1′‐acetoxychavicol acetate and sodium butyrate

Abstract

Introduction1′‐acetoxychavicol acetate (ACA) naturally obtained from the rhizomes and the seeds of the Zingiberaceae plant such as Languas galangal and Alpinia galangal. Butyric acid is one of major products of gut fermentation. Though ACA and sodium butyrate respectively upregulated phase II enzymes in intestinal epithelial cells (IEC 6), the interaction between these compounds is not clear. The present study was conducted to examine the interaction between ACA and sodium butyrate on regulation of phase II enzyme activities in IEC 6.MethodsGlutathione S‐transferase (GST) activity and NAD(P)H:quinine oxidoreductase 1 (NQO1) activity were measured by spectrophotometer. Nrf2 and p53 mRNA levels were estimated by RT‐PCR. Nrf2, p53, AMPK protein levels were examined by Western blotting.ResultsACA and sodium butyrate synergistically induced GST and NQO1 activities. These synergistic effects were inhibited with AMPK inhibitor, Compound C. Sodium butyrate upregulated the expression of Nrf2 mRNA and downregulated that of p53 mRNA, but ACA didn't affect those mRNA levels. The protein levels of intranuclear Nrf2 were increased by ACA or sodium butyrate. However, the combined treatment of ACA and sodium butyrate did not caused a synergic effect on level of intranuclar Nrf2. Intranuclar p53 protein level was increased by ACA, but It was decreased by sodium butyrate alone or by combined treatment of ACA and sodium butyrate. pAMPK levels were increased by combined treatment of ACA and sodium butyrate.ConclusionAMPK plays an important role on synergistic upregulation of phase II enzyme activities by ACA and sodium butyrate.Grant Funding Source : ASN