Calycosin-7-O-β-Glucoside Isolated from Astragalus membranaceus Promotes Osteogenesis and Mineralization in Human Mesenchymal Stem Cells

Abstract

Stem cells have received attention in various diseases, such as inflammatory, cancer, and bone diseases. Mesenchymal stem cells (MSCs) are multipotent stem cells that are critical for forming and repairing bone tissues. Herein, we isolated calycosin-7-O-β-glucoside (Caly) from the roots of Astragalus membranaceus, which is one of the most famous medicinal herbs, and investigated the osteogenic activities of Caly in MSCs. Caly did not affect cytotoxicity against MSCs, whereas Caly enhanced cell migration during the osteogenesis of MSCs. Caly increased the expression and enzymatic activities of ALP and the formation of mineralized nodules during the osteogenesis of MSCs. The osteogenesis and bone-forming activities of Caly are mediated by bone morphogenetic protein 2 (BMP2), phospho-Smad1/5/8, Wnt3a, phospho-GSK3β, and phospho-AKT, inducing the expression of runt-related transcription factor 2 (RUNX2). In addition, Caly-mediated osteogenesis and RUNX2 expression were attenuated by noggin and wortmannin. Moreover, the effects were validated in pre-osteoblasts committed to the osteoblast lineages from MSCs. Overall, our results provide novel evidence that Caly stimulates osteoblast lineage commitment of MSCs by triggering RUNX2 expression, suggesting Caly as a potential anabolic drug to prevent bone diseases.

The Herbal Combination of Radix astragali, Radix angelicae sinensis, and Caulis lonicerae Regulates the Functions of Type 2 Innate Lymphocytes and Macrophages Contributing to the Resolution of Collagen-Induced Arthritis

“…It has identified seven ingredients in R. astragali, six ingredients R. angelicae sinensis and eleven ingredients in C. lonicerae. Calycosin-7-O-β-glucoside, a bioactive compound isolated from R. astragali, has biological effects on triggering the processes of bone formation and repair (Park et al, 2021). Moreover, Ononin has been shown to induce apoptosis and reduce inflammation of FLSs in RA (Meng et al, 2021).…”

Effects of Triterpene Soyasapogenol B from Arachis hypogaea (Peanut) on Differentiation, Mineralization, Autophagy, and Necroptosis in Pre-Osteoblasts

“…Pre-osteoblast MC3T3E-1 cells (#CRL-2593, American-type culture collection, Manassas, VA, USA) were cultured at 37 °C in a humidified atmosphere of 5% CO 2 and 95% air using α-minimum essential medium (α-MEM) (WELGEME, Inc., Gyeonggido, Korea) without L-ascorbic acid (L-AA) (Sigma-Aldrich, St. Louis, MO, USA) and supplemented with 10% fetal bovine serum and 1 X Gibco ® Antibiotic-Antimycotic (Thermo Fisher Scientific, Waltham, MA, USA), as previously described [ 48 ]. The osteoblast differentiation of MC3T3E-1 cells was induced by changing to an osteogenic supplement medium (OS) containing 50 μg/mL L-ascorbic acid (L-AA) and 10 mM β-glycerophosphate (β-GP), as previously described [ 48 ]. The OS was replaced every 2 days during the incubation period.…”

“…For the early osteogenic activity analysis, the osteoblast differentiation of MC3T3-E1 cells was induced for 7 days, and alkaline phosphatase (ALP) staining and activity tests were performed, as previously described [ 48 ]. Briefly, for the ALP staining assay, the cells were incubated for 1 h at 37 °C with an ALP reaction solution (Takara Bio Inc., Tokyo, Japan) and the level of ALP staining was observed using a scanner and a light microscope.…”

“…Briefly, for the ALP staining assay, the cells were incubated for 1 h at 37 °C with an ALP reaction solution (Takara Bio Inc., Tokyo, Japan) and the level of ALP staining was observed using a scanner and a light microscope. To conduct the ALP activity assay as previously described [ 48 ], an alkaline phosphatase activity colorimetric assay kit (Biovision, Milpitas, CA, USA) was used and the ALP activity was quantitatively detected at 405 nm using the Multiskan GO Microplate Spectrophotometer (Thermo Fisher Scientific).…”

“…For the late osteogenic activity analysis, the differentiation was induced for 21 days, and an Alizarin red S (ARS) staining assay was performed, as previously described [ 48 ]. Briefly, cells were stained with 2% Alizarin red S (pH 4.2) (Sigma-Aldrich) for 10 min, then ARS staining was observed using a scanner and a light microscope, and then the staining level was quantitatively detected at 590 nm using the Multiskan GO Microplate Spectrophotometer (Thermo Fisher Scientific).…”