Potential of delphinidin-3-rutinoside extracted from Solanum melongena L. as promoter of osteoblastic MC3T3-E1 function and antagonist of oxidative damage
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Increasing evidence suggests the potential use of natural antioxidant compounds in the prevention/treatment of osteoporosis. This study was undertaken to investigate the effects of purified delphinidin-3-rutinoside (D3R), isolated from Solanum melongena L., on osteoblast viability and differentiation in basal conditions and its ability to protect MC3T3-E1 cells against oxidative damage induced by tert-butyl hydroperoxide (t-BHP).
MC3T3-E1 osteoblastic cells were treated with D3R (10−11–10−5 M for 24 h), followed by treatment with t-BHP (250 µM for 3 h). To test cell viability, MTT test was performed. Apoptotic cells were stained with Hoechst-33258 dye. Cytoskeleton rearrangement was stained with FICT-labelled phalloidin. Intracellular ROS production was measured using dichlorofluorescein CM-DCFA. The reduced glutathione to oxidized glutathione ratio (GSH/GSSG) contents was measured according to the OPT fluorimetric assay.
D3R (10−9 M) significantly increases viability of MC3T3-E1 cells and promotes osteoblast differentiation by increasing the expression of type I collagen, alkaline phosphatase and osteocalcin. Pre-treatment with D3R (10−9 M) significantly prevented t-BHP-induced osteoblastic dysfunction and changes in the cytoskeleton organization by decreasing intracellular ROS and preventing the reduction in GSH/GSSG. D3R did not significantly modify the expression of Osteoprotegerin/RANKL system activated by t-BHP suggesting a lack of effect of D3R on osteoblast/osteoclast crosstalk. D3R protective effects against t-BHP-induced osteoblastic dysfunction were mediated by the PI3K/Akt pathway since they were completely prevented by LY294002, a PI3K/Akt specific inhibitor.
These findings indicate that D3R protects MC3T3-E1 cells from oxidative damage and suggest the potential utility of dietary D3R supplement to prevent osteoblast dysfunction in age-related osteoporosis.
KeywordsDelphinidin-3-rutinoside Functional food component Oxidative stress Osteoblastic MC3T3-E1 cells
This work was supported by funds from PROGETTO CARIPLO GIOVANI 2015−0834 to Lavinia Casati. The authors thank the expertise and technical support of Dr. Giuseppe L. Rotino (CREA-ORL, Montanaso Lombardo) for providing the aubergine fruits and Prof. Giovanna Speranza (Dipartimento di Chimica, Università degli Studi di Milano) for the analysis by 1H-NMR of D3R crystals purity.
Compliance with ethical standards
Conflict of interest
The authors declare no conflicts of interest.
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