Isolation and Characterisation of Anti-diabetic Pharmacological Activities of Phytoestrogens

Abstract

Background: Isoflavones class of phytoestrogens including, genestein, daidzein and formononetin found in human dietary and show wide range of biological effects. These plant derived compounds have been shown to play a beneficial role in obesity and diabetes. Objective: In this study the impact of these phytoestrogens on glucose uptake in HepG2 cell were compared. Methods: Glucose uptake measurement was performed using 2-(N-(7-nitrobenzin-2-oxa-1, 3- diazol-4-yl) amino)-2-deoxyglucose (2-NBDG) and Omega FluoStar plate reader. Incubation of cells (104/ml, in 24 well plate at 37 °C in 5% CO2 / air) with three phytoestrogens at concentration of 10-4 M to 10-9M in two studies mood, short term treatment (one hour) and long term treatment (24 hours) was tested. Results: The data revealed, daidzein stimulates uptake of glucose, with a greater effect after a short treatment of one hour compared with treatment 24 hours. genistein exerted slightly inhibitory effect after one hour treatment compared with control, with the exception of treatment at 1 µM, which stimulated uptake about three-fold compared with control. Longer treatments with 10-4M to 10-6 M genistein resulted in gradual increase in glucose uptake to 2.4 times more than control, and thereafter a decline. A short treatment with formononetin inhibited glucose uptake, while longer treatments had variable effects, with an approximately two fold stimulation across a range of concentrations Conclusion: Overall HepG2 cells showed a significant increase in glucose uptake after treatment with phytoestrogens compared to the control. There was significant difference in glucose uptake between short and long term treatments, as indicated by two-way ANOVA.

Keywords

• phytoestrogens
• hyperglycemia
• <i> in vitro</i> study
• 2-nbdg