Abstract
BackgroundLeonurine, a major bioactive component from Herba leonuri, has been shown to exhibit anti-inflammatory and antioxidant effects. The aim of this study was to investigate the effect of leonurine on bone marrow-derived mesenchymal stem cells (BMSCs) as a therapeutic approach for treating osteoporosis.Materials and MethodsRat bone marrow-derived mesenchymal stem cells (rBMSCs) were isolated from 4-weeks-old Sprague–Dawley rats. The cytocompatibility of leonurine on rBMSCs was tested via CCK-8 assays and flow cytometric analyses. The effects of leonurine on rBMSC osteogenic differentiation were analyzed via ALP staining, Alizarin red staining, quantitative real-time polymerase chain reaction (qRT-PCR), and Western blot. Additionally, autophagy-related markers were examined via qRT-PCR and Western blot analyses of rBMSCs during osteogenic differentiation with leonurine and with or without 3-methyladenine (3-MA) as an autophagic inhibitor. Finally, the PI3K/Akt/mTOR signaling pathway was evaluated during rBMSC osteogenesis.ResultsLeonurine at 2–100 μM promoted the proliferation of rBMSCs. ALP and Alizarin red staining results showed that 10 μM leonurine promoted rBMSC osteoblastic differentiation, which was consistent with the qRT-PCR and Western blot results. Compared with those of the control group, the mRNA and protein levels of Atg5, Atg7, and LC3 were upregulated in the rBMSCs upon leonurine treatment. Furthermore, leonurine rescued rBMSC autophagy after inhibition by 3-MA. Additionally, the PI3K/AKT/mTOR pathway was activated in rBMSCs upon leonurine treatment.ConclusionLeonurine promotes the osteoblast differentiation of rBMSCs by activating autophagy, which depends on the PI3K/Akt/mTOR pathway. Our results suggest that leonurine may be a potential treatment for osteoporosis.
Highlights
Osteoporosis is the most common bone disease and is found in one-tenth of the population aged above 50 years and onequarter of the population aged above 80 years (Wright et al, 2014)
To assess the cytotoxicity of leonurine and its effect on the proliferation of bone marrow mesenchymal stem cells (BMSCs), we conducted a series of CCK-8 cell viability assays, cell cycle distribution analyses, and apoptosis assays with flow cytometry
Viability was significantly increased in the BMSCs treated with leonurine at a range of 2–100 μM, with a concentration of 10 μM resulting in a peak increase in viability followed by a gradual decrease at higher concentrations (40–100 μM)
Summary
Osteoporosis is the most common bone disease and is found in one-tenth of the population aged above 50 years and onequarter of the population aged above 80 years (Wright et al, 2014). The rate of occurrence of osteoporosisrelated bone fractures is increasing; it is estimated that one in two women and one in five men aged above 50 years are at risk of bone fracture as a direct result of osteoporosis (Das and Crockett, 2013). This condition is a major clinical and public health concern, and high morbidity, disability, and mortality rates are associated with the disease. The aim of this study was to investigate the effect of leonurine on bone marrow-derived mesenchymal stem cells (BMSCs) as a therapeutic approach for treating osteoporosis
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