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.
- 2.Almeida M, Han L, Martin-Millan M, Plotkin LI, Stewart SA, Roberson PK, Kousteni S, O’Brien CA, Bellido T, Parfit AM, Weinstein RS, Jilka RL, Manolagas SC (2007) Skeletal involution by age-associated oxidative stress and its acceleration by loss of sex steroids. J Biol Chem 282(37):27285–27297CrossRefGoogle Scholar
- 4.D’Amelio P, Cristofaro MA, Tamone C, Morra E, Di Bella S, Isaia G, Grimaldi A, Gennero L, Gariboldi A, Ponzetto A, Pescarmona GP, Isaia GC (2008) Role of iron metabolism and oxidative damage in postmenopausal bone loss. Bone. 43(6):1010–1015. https://doi.org/10.1016/j.bone.2008.08.107CrossRefGoogle Scholar
- 6.Jilka RL, Almeida M, Ambrogini E, Han L, Roberson PK, Weinstein RS, Manolagas SC (2010) Decreased oxidative stress and greater bone anabolism in the aged, when compared to the young, murine skeleton with parathyroid hormone administration. Aging Cell 9(5):851–867. https://doi.org/10.1111/j.1474-9726.2010.00616.xCrossRefGoogle Scholar
- 10.Moriwaki S, Suzuki K, Muramatsu M, Nomura A, Inoue F, Into T, Yoshiko Y, Niida S (2014) Delphinidin, one of the major anthocyanidins, prevents bone loss through the inhibition of excessive osteoclastogenesis in osteoporosis model mice. PLoS One 9(5):e97177. https://doi.org/10.1371/journal.pone.0097177CrossRefGoogle Scholar
- 22.Yi L, Chen CY, Jin X, Zhang T, Zhou Y, Zhang QY, Zhu JD, Mi MT (2012) Differential suppression of intracellular reactive oxygen species-mediated signaling pathway in vascular endothelial cells by several subclasses of flavonoids. Biochimie 94(9):2035–2044. https://doi.org/10.1016/j.biochi.2012CrossRefGoogle Scholar
- 23.He J, Giusti MM (2010) Anthocyanins: natural colorants with health-promoting properties. Annu Rev Food Sci Technol 1:163 – 87 doi. https://doi.org/10.1146/annurev.food.080708.100754Google Scholar
- 24.Jing P, Qian B, Zhao S, Qi X, Ye L, Mónica Giusti M, Wang X (2015) Effect of glycosylation patterns of Chinese eggplant anthocyanins and other derivatives on antioxidant effectiveness in human colon cell lines. Food Chem 172:183–189. https://doi.org/10.1016/j.foodchem.2014.08.100CrossRefGoogle Scholar
- 32.Dieci E, Casati L, Pagani F, Celotti F, Sibilia V (2014) Acylated and unacylated ghrelin protect MC3T3-E1 cells against tert-butyl hydroperoxide-induced oxidative injury: pharmacological characterization of ghrelin receptor and possible epigenetic involvement. Amino Acids 46(7):1715–1725CrossRefGoogle Scholar
- 33.Zhang JK, Yang L, Meng GL, Fan J, Chen JZ, He QZ, Chen S, Fan JZ, Luo ZJ, Liu J (2012) Protective effect of tetrahydroxystilbene glucoside against hydrogen peroxide-induced dysfunction and oxidative stress in osteoblastic MC3T3-E1 cells. Eur J Pharmacol 689(1–3):31–37. https://doi.org/10.1016/j.ejphar.2012.05.045CrossRefGoogle Scholar
- 39.Farley JR, Hall SL, Tanner MA, Wergedal JE (1994) Specific activity of skeletal alkaline phosphatase in human osteoblast-line cells regulated by phosphate, phosphate esters, and phosphate analogs and release of alkaline phosphatase activity inversely regulated by calcium. J Bone Miner Res 9(4):497–508CrossRefGoogle Scholar
- 41.Rodan GA, Noda M (1991) Gene expression in osteoblastic cells. Crit Rev Eukaryot Gene Expr 1(2):85–98Google Scholar
- 49.Wang B, Shravah J, Luo H, Raedschelders K, Chen DD, Ansley DM (2009) Propofol protects against hydrogen peroxide-induced injury in cardiac H9c2 cells via Akt activation and Bcl-2 up-regulation. Biochem Biophys Res Commun 389(1):105–111. https://doi.org/10.1016/j.bbrc.2009.08.097CrossRefGoogle Scholar
- 52.Sunters A, Armstrong VJ, Zaman G, Kypta RM, Kawano Y, Lanyon LE, Price JS (2010) Mechano-transduction in osteoblastic cells involves strain-regulated estrogen receptor alpha-mediated control of insulin-like growth factor (IGF) I receptor sensitivity to Ambient IGF, leading to phosphatidylinositol 3-kinase/AKT-dependent Wnt/LRP5 receptor-independent activation of beta-catenin signaling. J Biol Chem 285(12):8743–8758. https://doi.org/10.1074/jbc.M109.027086CrossRefGoogle Scholar
- 55.Yi L, Chen CY, Jin X, Mi MT, Yu B, Chang H, Ling WH, Zhang T (2010) Structural requirements of anthocyanins in relation to inhibition of endothelial injury induced by oxidized low-density lipoprotein and correlation with radical scavenging activity. FEBS Lett 584(3):583–90. https://doi.org/10.1016/j.febslet.2009.12.006.CrossRefGoogle Scholar
- 59.Reuter S, Gupta SC, Chaturvedi MM, Aggarwal BB (2010) Oxidative stress, inflammation, and cancer: How are they linked? Free Radic Biol Med 49(11):1603–1616. https://doi.org/10.1016/j.freeradbiomed.2010.09.006CrossRefGoogle Scholar
- 61.Liu H, Mao P, Wang J, Wang T, Xie CH2 (2016) Azilsartan, an angiotensin II type 1 receptor blocker, attenuates tert-butyl hydroperoxide-induced endothelial cell injury through inhibition of mitochondrial dysfunction and anti-inflammatory activity. Neurochem Int 94:48–56. https://doi.org/10.1016/j.neuint.2016.02.005CrossRefGoogle Scholar
- 62.Lee SG, Kim B, Yang Y, Pham TX, Park YK, Manatou J, Koo SI, Chun OK, Lee JY (2014) Berry anthocyanins suppress the expression and secretion of proinflammatory mediators in macrophages by inhibiting nuclear translocation of NF-κB independent of NRF2-mediated mechanism. J Nutr Biochem 25(4):404–411. https://doi.org/10.1016/j.jnutbio.2013.12.001CrossRefGoogle Scholar