Abstract
ObjectiveWe aimed to investigate the therapeutic effects of Moringa oleifera leaf extracts on osteogenic induction of rat bone marrow mesenchymal stem cells (BMSCs) following peroxidative damage and to explore the underlying mechanisms.MethodsConditioned medium was used to induce osteogenic differentiation of BMSCs, which were treated with H2O2, Moringa oleifera leaf extracts-containing serum, or the phosphatidyl inositol-3 kinase (PI3K) inhibitor wortmannin, alone or in combination. Cell viability was measured using the MTT assay. Cell cycle was assayed using flow cytometry. Expression levels of Akt, phosphorylated (p)Akt, Foxo1, and cleaved caspase-3 were analyzed using western blot analysis. The mRNA levels of osteogenesis-associated genes, including alkaline phosphatase (ALP), collagen І, osteopontin (OPN), and Runx2, were detected using qRT-PCR. Reactive oxygen species (ROS) and malondialdehyde (MDA) levels, as well as superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), and ALP activity were detected using commercially available kits. Osteogenic differentiation capability was determined using alizarin red staining.ResultsDuring osteogenic induction of rat BMSCs, H2O2 reduced cell viability and proliferation, inhibited osteogenesis, increased ROS and MDA levels, and decreased SOD and GSH-PX activity. H2O2 significantly reduced pAkt and Foxo1 expression, and increased cleaved caspase-3 levels in BMSCs. Additional treatments with Moringa oleifera leaf extracts partially reversed the H2O2-induced changes. Wortmannin partially attenuated the effects of Moringa oleifera leaf extracts on protein expression of Foxo1, pAkt, and cleaved caspase-3, as well as mRNA levels of osteogenesis-associated genes.ConclusionMoringa oleifera leaf extracts ameliorate peroxidative damage and enhance osteogenic induction of rat BMSCs by activating the PI3K/Akt/Foxo1 pathway.
Highlights
Aging can cause peroxidative damage, which can lead to decreased bone mass and osteoporosis [1], which is closely related to frailty [2], increasing the risk of negative events such as disability, paralysis, and tumble [3]
Moringa oleifera leaf extracts containing serum were used to test the protective effects on Bone marrow mesenchymal stem cell (BMSC) challenged with peroxidative damage during osteogenic differentiation, and we explored the possible cellular and molecular mechanisms underlying the protective properties of Moringa oleifera by using wortmannin [28]
We evaluated the impact of Moringa oleifera on attenuating dysregulated cell viability and cell cycle progression of BMSCs treated with 100 μmol/L Hydrogen peroxide (H2O2)
Summary
Aging can cause peroxidative damage, which can lead to decreased bone mass and osteoporosis [1], which is closely related to frailty [2], increasing the risk of negative events such as disability, paralysis, and tumble [3]. Bone marrow mesenchymal stem cells (BMSCs) are a subgroup of cells with multi-differentiation potential, and their directed differentiation is a complex process involving multiple cellular pathways. BMSCs with multi-differentiation properties are currently used to treat a variety of diseases, such as type 1 diabetes, inflammatory bowel disease, and cardiac disease [9,10,11]. Several studies have demonstrated that BMSCs can be induced to differentiate into bone cells under specific induction conditions [12, 13]. Studies in animal models have revealed that both allogeneic and autologous BMSC transplantation is applicable in the treatment of osteoporosis [14]. Accumulating evidence has confirmed that transplantation of BMSCs can promote formation of new bones, improve bone quality, and prevent osteoporosis-associated bone fractures [15,16,17]
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