Related Experiment Video
Updated: Nov 16, 2025
Isolation of Mesenchymal Stem Cells from Human Alveolar Periosteum and Effects of Vitamin D on Osteogenic Activity of Periosteum-derived Cells
Published on: May 4, 2018
Moringa oleifera leaf extracts protect BMSC osteogenic induction following peroxidative damage by activating the
Meiling Liu1, Haifeng Ding1, Hongzhi Wang1
1Department of Geriatrics, Second Affiliated Hospital of Harbin Medical University, 246 Xuefu Road, Nangang District, Harbin, 150086, China.
Objective:
We 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.
Methods:
Conditioned 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.
Results:
During 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.
Conclusion:
Moringa oleifera leaf extracts ameliorate peroxidative damage and enhance osteogenic induction of rat BMSCs by activating the PI3K/Akt/Foxo1 pathway.
Insights
Moringa oleifera leaf extracts protect rat bone marrow mesenchymal stem cells (BMSCs) from oxidative damage and promote bone formation. This occurs by activating the PI3K/Akt/Foxo1 pathway, crucial for osteogenic induction.
Area of Science:
- Stem cell biology
- Biochemistry
- Pharmacology
Background:
- Oxidative stress impairs bone marrow mesenchymal stem cells (BMSCs) osteogenic differentiation.
- Moringa oleifera leaf extracts (MOLE) possess potential therapeutic properties.
Purpose of the Study:
- To evaluate MOLE's effects on osteogenic induction of BMSCs under peroxidative damage.
- To elucidate the underlying molecular mechanisms involving the PI3K/Akt/Foxo1 pathway.
Main Methods:
- BMSCs were treated with hydrogen peroxide (H2O2), MOLE, and a PI3K inhibitor (wortmannin).
- Assays included MTT for viability, flow cytometry for cell cycle, Western blot for protein expression (Akt, pAkt, Foxo1, caspase-3), qRT-PCR for gene expression (ALP, Collagen I, OPN, Runx2), and ROS/MDA/SOD/GSH-PX activity measurements.
- Osteogenic differentiation was assessed via alizarin red staining.
Main Results:
- H2O2 reduced BMSCs viability, proliferation, and osteogenesis, while increasing oxidative stress markers (ROS, MDA) and decreasing antioxidant enzymes (SOD, GSH-PX).
- H2O2 decreased pAkt and Foxo1, and increased cleaved caspase-3. MOLE treatment partially reversed these effects.
- Wortmannin partially blocked MOLE's impact on protein and gene expression, indicating PI3K pathway involvement.
Conclusions:
- Moringa oleifera leaf extracts mitigate oxidative damage in rat BMSCs.
- MOLE enhance osteogenic induction by activating the PI3K/Akt/Foxo1 signaling pathway.
