Purpose Natural antioxidants are considered as promising compounds in the prevention/treatment of osteoporosis. We studied the ability of purified δ-tocotrienol (δ-TT) isolated from a commercial palm oil (Elaeis guineensis) fraction to protect osteoblast MC3T3-E1 and osteocyte MLO-Y4 cells against tert-butyl hydroperoxide (t-BHP)-induced oxidative damage and the mechanisms involved in its protective action in MC3T3-E1. Methods MC3T3-E1 and MLO-Y4 cells were treated with δ-TT (1.25–20 µg/ml for 2 h) followed by t-BHP at 250 µM or 125 µM for 3 h, respectively. MTT test was used to measure cell viability. Apoptotic cells were stained with Hoechst-33258 dye. Intracellular ROS levels were measured by dichlorofluorescein CM-DCFA. The OPT fluorimetric assay was used to detect the reduced glutathione to oxidized glutathione ratio (GSH/GSSG) contents. Results δ-TT significantly prevented the effects of t-BHP on cell viability and apoptosis reaching a maximum protective activity at 10 and 5 µg/ml in MC3T3-E1 and MLO-Y4 cells, respectively. This protective effect was due to a reduction of intracellular ROS levels and an increase in the defense systems shown by the increase in the GSH/GSSG. GSH loss induced by an inhibitor of GSH synthesis significantly reduced the δ-TT-positive effect on ROS levels. δ-TT prevention of oxidative damage was completely removed by combined treatment with the specific inhibitors of PI3K/AKT (LY294002) and Nrf2 (ML385). Conclusions The δ-TT protective effect against oxidative damage in MC3T3-E1 cells is due to a reduction of intracellular ROS levels and an increase of the GSH/GSSG ratio, and involves an interaction between the PI3K/Akt–Nrf2 signaling pathways.
Beneficial effects of δ-tocotrienol against oxidative stress in osteoblastic cells: studies on the mechanisms of action / L. Casati, F. Pagani, P. Limonta, C. Vanetti, G. Stancari, V. Sibilia. - In: EUROPEAN JOURNAL OF NUTRITION. - ISSN 1436-6207. - (2019 Jul 06). [Epub ahead of print]
Titolo: | Beneficial effects of δ-tocotrienol against oxidative stress in osteoblastic cells: studies on the mechanisms of action |
Autori: | CASATI, LAVINIA (Primo) SIBILIA, VALERIA (Ultimo) (Corresponding) |
Parole Chiave: | Tocotrienol; Oxidative stress; MC3T3-E1 cells; MLO-Y4 cells; Signaling pathways |
Settore Scientifico Disciplinare: | Settore BIO/14 - Farmacologia Settore MED/04 - Patologia Generale Settore BIO/13 - Biologia Applicata |
Progetto: | Epigenetic in the pathophysiology of Osteoporosis: new insight from in vivo and in vitro studies. |
Data di pubblicazione: | 6-lug-2019 |
Rivista: | |
Tipologia: | Article (author) |
Data ahead of print / Data di stampa: | 6-lug-2019 |
Digital Object Identifier (DOI): | http://dx.doi.org/10.1007/s00394-019-02047-9 |
Appare nelle tipologie: | 01 - Articolo su periodico |
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