Resveratrol-Mediated Reversal of Doxorubicin-Induced Osteoclast Differentiation

Abstract

Secondary osteoporosis has been associated with cancer patients undertaking Doxorubicin (DOX) chemotherapy. However, the molecular mechanisms behind DOX-induced bone loss have not been elucidated. Molecules that can protect against the adverse effects of DOX are still a challenge in chemotherapeutic treatments. We investigated the effect and mechanism of DOX in osteoclast differentiation and used the Sirt 1 activator resveratrol (RES) to counteract DOX-induced effects. RAW 264.7 cells were differentiated into osteoclasts under cotreatment with DOX and RES, alone or combined. RES treatment inhibited DOX-induced osteoclast differentiation, reduced the expression of osteoclast fusion marker Oc-stamp and osteoclast differentiation markers Rank, Trap, Ctsk and Nfatc1. Conversely, RES induced the upregulation of antioxidant genes Sod 1 and Nrf 2 while DOX significantly reduced the FoxM1 expression, resulting in oxidative stress. Treatment with the antioxidant MitoTEMPO did not influence DOX-induced osteoclast differentiation. DOX-induced osteoclastogenesis was studied using the cathepsin-K zebrafish reporter line (Tg[ctsk:DsRed]). DOX significantly increased ctsk signal, while RES cotreatment resulted in a significant reduction in ctsk positive cells. RES significantly rescued DOX-induced mucositis in this model. Additionally, DOX-exposed zebrafish displayed altered locomotor behavior and locomotory patterns, while RES significantly reversed these effects. Our research shows that RES prevents DOX-induced osteoclast fusion and activation in vitro and in vivo and reduces DOX-induced mucositis, while improving locomotion parameters.

Revisiting Resveratrol as an Osteoprotective Agent: Molecular Evidence from In Vivo and In Vitro Studies

“…In RAW 264.7 cells exposed to H 2 O 2 , RSV improved oxidative stress status by increasing SOD and GSH-PX levels, while decreasing ROS and MDA levels [37]. RSV was also found to increase the expression of the antioxidant genes SOD 1 and Nrf2 in RAW264.7 cells exposed to doxorubicin [106]. RSV was also able to reduce the mRNA expression of the osteoclast fusion marker, Oc-stamp, as well as the osteoclast differentiation markers RANK, TRAP, and CtsK by regulating FOXO1 transcriptional activity and inhibiting the PI3K/AKT [37] and activated T-cells cytoplasmic 1 (NFATc1) [106] signalling pathways, which are important in osteoclastogenesis (Summarised in Table 2).…”

“…In the 3-D osteogenic differentiation on collagen scaffold of rat adipose stem cells treated with RSV, an increase in OPG was observed along with an increase in the mineralisation process [64]. In addition, RSV inhibited the expression of osteoclast markers (RANK, NFATc1, TRAP, CtsK, and Oc-Stamp) in RAW264.7 cells to suppress the generation of osteoclast-like cells [106]. In an in vivo study, RSV reduced the osteoclastic differentiation marker (TRAP and CTX-1) by inhibition of NF-κB and NFATc1 signalling pathway [45].…”

“…RSV treatment increased the expressions of FoxO1, SIRT1, and OPG genes while decreasing the expression of RANKL. In an in vitro study, RSV inhibited the NFATc1 signalling pathway to prevent doxorubicin-induced osteoclast fusion and activation and to increase FoxO1 transcriptional activity [106]. Another study found that RSV increased FoxO1 transcrip-tional activity by inhibiting the PI3K/AKT signalling pathway, promoting oxidative stress resistance, and inhibiting osteoclastogenesis [37].…”

The osteogenic and mineralogenic potential of the microalgae Skeletonema costatum and Tetraselmis striata CTP4 in fish models

“…Reactive oxygen species (ROS) are known to negatively affect bone mineral status and affect bone cells through several mechanisms, by inducing osteoclastic differentiation while suppressing osteoblastic differentiation and survival, being at the basis of various bone erosive pathologies including age-related osteoporosis [ 54 , 55 ]. In this regard, the exposure of mammalian and fish models (in vivo and in vitro) to pro-oxidant agents have been shown to inhibit osteoblastic differentiation and activity and increased the incidence of skeletal anomalies, but the supplementation of antioxidants could counteract these negative outputs [ 56 – 61 ]. Recent studies reporting the osteoactive and antioxidative potential of hydroethanolic, methanolic, and dichloromethane extracts prepared from green ( Cladophora rupestris and Codium fragile ) and red ( Ceramium secundatum , Ceramium pallidum , and Plocamium lyngbyanum ) macroalgae, further support this hypothesis [ 5 , 6 , 62 ].…”

Nutraceuticals and Functional Foods: A Comprehensive Review of Their Role in Bone Health

“…In detail, it has been reported that it reduces the expression of bone resorption markers in RAW264.7 cells such as cathepsin K and MMP9. Resveratrol (10 −5 M) decreases the expression of the OC fusion marker Oc-stamp, RANK, TRAP, and cathepsin K by regulating the transcriptional activity of FOXO1, the PI3K/AKT [ 37 ] and NFATc1 signaling pathways [ 38 ]; see Figure 2 .…”