Cajanolactone A from Cajanus cajan Promoted Osteoblast Differentiation in Human Bone Marrow Mesenchymal Stem Cells via Stimulating Wnt/LRP5/β-Catenin Signaling

Abstract

Cajanolactone A (CLA) is a stilbenoid discovered by us from Cajanus cajan (L.) Millsp. In our study, CLA was found to promote osteoblast differentiation in human bone marrow mesenchymal stem cells (hBMSCs), as judged by increased cellular alkaline phosphatase activity and extracellular calcium deposits, and elevated protein expression of Runx2, collagen-1, bone morphogenetic protein-2, and osteopontin. Mechanistic studies revealed that hBMSCs undergoing osteoblast differentiation expressed upregulated mRNA levels of Wnt3a, Wnt10b, LRP5/6, Frizzled 4, β-catenin, Runx2, and Osterix from the early stage of differentiation, indicating the role of activated Wnt/β-catenin signaling pathway in osteoblast differentiation. Addition of CLA to the differentiation medium further increased the mRNA level of Wnt3a, Wnt10b, Frizzled 4, LRP5, and β-catenin, inferring that CLA worked by stimulating Wnt/LRP5/β-catenin signaling. Wnt inhibitor dickkopf-1 antagonized CLA-promoted osteoblastogenesis, indicating that CLA did not target the downstream of canonical Wnt signaling pathway. Treatment with CLA caused no changes in mRNA expression level, as well as protein secretion of osteoprotegerin (OPG) and receptor activator of nuclear factor kappa-B ligand (RANKL), indicating that CLA did not affect the OPG/RANKL axis. Our results showed that CLA, which promoted osteoblast differentiation in hBMSCs, through activating Wnt/LRP5/β-catenin signaling transduction, is a promising anti-osteoporotic drug candidate.

Four new isocoumarins from Cajanus cajan

“…To the best of our knowledge, the only isocoumarin isolated from C. cajan was named cajanolactone A (CLA), which was found to promote osteoblast differentiation in hBMSCs through activating Wnt/LRP5/b-catenin signaling transduction. 13 Our ongoing search for more structurally diverse compounds from C. cajan resulted in the isolation of four new isocoumarins (1-4) (Fig. 1), the isolation, structural elucidation of which are reported in the present report.…”

“…The 1 H-NMR and 13 C-NMR spectra ( Analyses of the 1D and 2D NMR data of 3 and 2 revealed their close resemblance except for the positions of the free hydroxyl group and the isoprenyl moiety in 3, which was further assigned by the observed HMBC cross-peaks of 7-OH to C-1 and C-8, and H 2 -1 00 to C-3 and C-4, respectively. Detailed inspection of 2D NMR (HSQC, 1 H-1 H COSY, HMBC) spectra established the planar structure of 3 as depicted in Fig.…”

“…Compound 4 was also isolated as a yellowish oil, which exhibited an ion peak at m/z 355.1531 [M + H] + (calcd 355.1540) 2 1 H (500 MHz) and 13 in HRESIMS spectrum with a positive mode, giving the molecular formula of C 21 H 22 O 5 with eleven degrees of unsaturation. The planar structure of 4 was determined to be the same as 3 based on a comparison of 1D and 2D NMR data.…”

Components identification and isomers differentiation in pigeon pea (Cajanus cajan L.) leaves by LC‐MS

“…Hence, compound 46 was tentatively identified as cajanuslactone [24]. Compound 50 without efficient fragment ions was preliminary assigned as cajanolactone A according to the previous report [25].…”

Osteoblastgenic and Osteogenic Effects of KY-273 with CDK8/19 Inhibitory Activity in Bone Marrow Mesenchymal Stem Cells and Female Rats

“…Orally active osteogenic drugs have been a desirable target, and a great number of small molecules have been reported to exert in vitro osteoblastogenic effects and in vivo osteogenic effects. [1][2][3][4][5][6] Unfortunately, these have not been yet developed as effective and safe anti-osteoporotic drugs. Among them, benzothiopyran, benzofuran, and coumarin derivatives are suggested to enhance osteoblast differentiation via BMP potentiation.…”