Supplementation with Fish Oil and Genistein, Individually or in Combination, Protects Bone against the Adverse Effects of Methotrexate Chemotherapy in Rats

Abstract

Cancer chemotherapy has been shown to induce long-term skeletal side effects such as osteoporosis and fractures; however, there are no preventative treatments. This study investigated the damaging effects of anti-metabolite methotrexate (MTX) subcutaneous injections (0.75 mg/kg BW) for five days and the potential protective benefits of daily oral gavage of fish oil at 0.5 mL/100 g BW (containing 375 mg of n-3 PUFA/100 g BW), genistein (2 mg/100 g BW), or their combination in young adult rats. MTX treatment alone significantly reduced primary spongiosa height and secondary spongiosa trabecular bone volume. Bone marrow stromal cells from the treated rats showed a significant reduction in osteogenic differentiation but an increase in adipogenesis ex vivo. Consistently, stromal cells had significantly higher mRNA levels of adipogenesis-related proliferator activator activated receptor-γ (PPAR-γ) and fatty acid binding protein (FABP4). MTX significantly increased the numbers of bone-resorbing osteoclasts and marrow osteoclast precursor cell pool while significantly enhancing the mRNA expression of receptor activator for nuclear factor kappa B ligand (RANKL), the RANKL/osteoprotegerin (OPG) ratio, interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in the bone. Supplementary treatment with fish oil and/or genistein significantly preserved trabecular bone volume and osteogenesis but suppressed MTX-induced adipogenesis and increases in osteoclast numbers and pro-osteoclastogenic cytokine expression. Thus, Fish oil and/or genistein supplementation during MTX treatment enabled not only preservation of osteogenic differentiation, osteoblast number and bone volume, but also prevention of MTX treatment-induced increases in bone marrow adiposity, osteoclastogenic cytokine expression and osteoclast formation, and thus bone loss.

The Protective Role of Simvastatin on Methotrexate-Induced Bone Injury in Adult Albino Rat

“…Reiner and Bertha described this phenomenon as if the trabeculae were formed of small islands of bone [35] . Also, MTX therapy might lead to overall reduction of trabecular bone volume [36] . Experimental studies of MTX induced bone defects revealed that apart from the reduced osteoblast number and trabecular bone volume; there was a significant increase in marrow adiposity [31,37] .…”

“…In addition, due to increased usage of anti-cancer drugs and their significant effect on skeletal health, it becomes important to explore potential supplementary treatments that may be useful in protecting bone during cancer chemotherapy [14] . Folinic acid, soy products rich in isoflavone, genistein, and fish oils rich in omega-3 polyunsaturated fatty acids were previously used to improve bone health [15][16][17][18] .…”

Combination chemotherapy with cyclophosphamide, epirubicin and 5-fluorouracil causes trabecular bone loss, bone marrow cell depletion and marrow adiposity in female rats

“…Following fixation, cells were stained in Nile Red to identify adipogenic colonies under fluorescence microscope (excitation, 485 nm; emission, 525 nm). Adipogenic differentiation potential was expressed as the percentage of positive colonies over the total toluidine blue-stained colonies as described [22].…”

Steamed root of Rehmannia glutinosa Libosch (Plantaginaceae) alleviates methotrexate-induced intestinal mucositis in rats

“…Both DNA and non-DNA damages exert on epithelial and submucosal cells following chemotherapy, which cause intricate sequential biological events, such as generation of reactive oxygen species (ROS), transcription factor activation, pro-inflammatory cytokine expression/accumulation, nitrosative stress, cell apoptosis, reduction in epithelial cell proliferation, changes in growth factor expression, proteolysis, bacterial colonization, and activation of enteric immune and nervous systems (Al-Dasooqi et al, 2013;Kolli et al, 2008;Logan et al, 2007;2009;Qutob et al, 2013;Sonis, 2009;Xian et al, 1999Xian et al, , 2000Xian et al, , 2002Xian, 2003;Zhan et al, 2014). Great efforts have been applied to evaluate/ develop interventions that prevent or reduce the toxic effects of chemotherapy, such as intensive oral hygiene care and usage of antiseptics, antimicrobial and anti-inflammatory agents, cytokines, growth factors, antioxidants, immune modulators and homeopathic agents (McCulloch et al, 2014;Nadhanan et al, 2013;Qutob et al, 2013;Rodríguez-Caballero et al, 2012).…”