Hypoglycemic activity and constituents analysis of blueberry (Vaccinium corymbosum) fruit extracts

Background: To investigate hypoglycemic activity and elucidate the active composition of the fruit blueberry (Vaccinium corymbosum). Methods: Methanol extracts of blueberry (MEB) were separated using a D101 macroporous resin column to yield quinic acid derivative (Fr.1)- and flavonoid (Fr.2)-rich fractions. The effects of the blueberry extracts on mRNA expression of GLUT-2 (glucose transporter type 2) and PPAR gamma(peroxisome proliferator-activated receptor-gamma), as well as on the activities of PPRE (peroxisome proliferator response element) and NF-kappa B were analyzed in LO2 normal liver cells. Real-time PCR was used to detect the expression of GLUT-2, PPAR gamma, TNF-alpha, IL-1 beta, and IL-6 mRNA. The PPRE and NF-kappa B activities were detected by a luciferase reporter assay. Western blotting was used to detect the levels of PPAR gamma, GLUT-2, and p65. The active compositions were isolated using various chromatography columns, and were analyzed by NMR. Results: mRNA and protein expression of GLUT-2 and PPAR gamma were significantly increased upon treatment with 400 mu g/mL extracts of blueberry (P<0.05). The PPRE activity was also significantly increased in a dose-dependent manner upon administration of MEB (P<0.05). Furthermore, the NF-kappa B activity induced by lipopolysaccharides was inhibited by MEB (P< 0.05). No fraction separated from MEB exhibited PPRE activation or NF-kappa B inhibition activity. Blueberry extract may execute its hypoglycemic activity by stimulating expression of GLUT-2 and PPAR gamma, and by inhibiting the inflammatory pathway. Together, quinic acid derivatives and flavonoids may result in a synergistic effect. Fourteen phenolic acids, including eight flavonoids, four quinic acid derivatives, and two other phenolic acids, were isolated and identified, and caffeoylquinic acid derivatives and quercetin glycosides were found to be the major constituents of blueberry. Conclusion: Blueberry may have hypoglycemic activity that functions through synergistic effects with caffeoylquinic acid derivatives and quercetin glycosides.