Cottonseed Extracts and Gossypol Regulate Diacylglycerol Acyltransferase Gene Expression in Mouse Macrophages.

Plant bioactive polyphenols have been used for the prevention and treatment of various diseases since ancient history. Cotton ( Gossypium hirsutum L.) seeds are classified as glanded or glandless depending on the presence or absence of pigment glands, which contain polyphenolic gossypol. Diacylglycerol acyltransferases (DGATs) are integral membrane proteins that catalyze the last step of triacylglycerol biosynthesis in eukaryotes. Understanding the regulation of DGATs will provide information for therapeutic intervention for obesity and related diseases. However, little was known if DGAT gene expression was regulated by natural products. The objective of this study was to investigate the effects of cottonseed extracts and gossypol on DGAT gene expression in mouse RAW264.7 macrophages. Mouse cells were treated with different concentrations of cottonseed extracts, gossypol, and lipopolysaccharides (LPS) for various times. Quantitative polymerase chain reaction assay showed that coat extract of glanded seeds had a modest effect on DGAT1 and minimal effect on DGAT2 mRNA levels. Kernel extract of glanded seeds had a minimal effect on DGAT1 but increased DGAT2 mRNA levels more than 20-fold. Coat extract of glandless seeds and LPS had minimal effects on DGAT mRNA levels. Kernel extract of glandless seeds did not have much effect on DGAT1 and slightly increased DGAT2 mRNA levels. Gossypol increased DGAT1 and DGAT2 mRNA levels by up to three-fold and more than 80-fold, respectively. The coefficient correlations ( R2) between DGAT2 mRNA levels and glanded kernel extract and gossypol concentrations were 0.82-0.99. This study suggests that Dgat2 is an inducible gene rapidly responding to stimulators such as polyphenols whose protein product DGAT2 plays an important role in fat biosynthesis. We conclude that gossypol and ethanol extract from glanded cottonseed kernel are strong stimulators of DGAT2 gene expression and that they may be novel agents for intervention of lipid-related dysfunction via increasing DGAT2 gene expression in target tissues.