Analytical Methods and Bioassays for Cytotoxicity and Antidiabetic Properties of Aquilaria crassna Leaf Extracts in HepG2 Cells

Abstract

Aquilaria crassna is a herbal plant that has recently been reported to possess several biological activities. A. crassna leaf extracts have been demonstrated to have a glucose-lowering effect in animal models. However, it is unclear what phytochemical compounds mediate this antidiabetic property. Here, we describe analytical methods for identifying such compounds from dried leaves by differential extractions with ethanol, butanol, ethyl acetate, and water, respectively. The phytochemical compounds in each fraction were further identified by gas chromatography-mass spectrometry. The cytotoxicity of these fractions was tested against a HepG2 cell line, while the rate of glucose utilization was determined using glucose oxidase assay. Lastly, the inhibitory effect on suppression of hepatic glucose production in HepG2 cells was determined by quantitative real-time PCR of genes encoding pyruvate carboxylase, phosphoenolpyruvate carboxykinase, fructose-1,6-bisphosphatase, glucose-6-phosphatase, and liver glycogen synthase.