Oridonin promotes osteogenesis through Wnt/β-catenin pathway and inhibits RANKL-induced osteoclastogenesis in vitro
| Autor: | Jiali Zhao, Huilin Yang, Yingkang Huang, Feng Kuai, Liang Zhou |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
musculoskeletal diseases
0301 basic medicine Cell Survival viruses Osteoclasts 030226 pharmacology & pharmacy General Biochemistry Genetics and Molecular Biology Rats Sprague-Dawley 03 medical and health sciences 0302 clinical medicine Western blot Osteoclast Osteogenesis medicine Animals General Pharmacology Toxicology and Pharmaceutics Wnt Signaling Pathway Cells Cultured biology medicine.diagnostic_test Dose-Response Relationship Drug Chemistry Mesenchymal stem cell RANK Ligand Wnt signaling pathway Osteoblast Cell Differentiation Mesenchymal Stem Cells General Medicine biochemical phenomena metabolism and nutrition Molecular biology Rats RUNX2 030104 developmental biology medicine.anatomical_structure RANKL Catenin biology.protein Diterpenes Kaurane |
| Zdroj: | Life sciences. 262 |
| ISSN: | 1879-0631 |
| Popis: | Aims To study the molecular mechanism of oridonin (ORI) on osteoblast differentiation and osteoclast formation in vitro. Main methods Rat bone marrow mesenchymal stem cells (BMSCs) were treated with different concentrations of ORI in osteogenic medium (OM). CCK-8 assay and were used to detect the effect on BMSCs viability. Alizarin red staining and ALP activity were used to illuminate the effect of ORI on osteogenic differentiation. Expressions of osteogenic differentiation related genes were detected by real-time quantitative PCR (qRT-PCR), and expressions of osteogenic related proteins were detected by Western blot (WB) and immunofluorescence. Similarly, bone marrow mononuclear cells (BMMs) were treated with different concentrations of ORI. CCK-8 assay and Live/Dead staining were used to detect the effect of ORI on BMMs activity. TRAP staining was used to detect its effect on osteoclast differentiation. Expressions of osteoclast-related genes were detected by qRT-PCR, and expressions of osteoclast-related proteins were detected by WB and immunofluorescence. Key findings (1) ORI (2 μM) promoted the ALP activity of BMSCs differentiation into osteoblasts and increased the number of calcium nodules. (2) ORI stimulated the expressions of wnt1, β-catenin and Runx2, but with no significantly effect on p-GSK-3β and GSK-3β. (3) ORI promoted the expression of OPG and inhibited the expression of RANKL. (4) ORI directly/indirectly inhibited the osteoclast formation and expressions of osteoclast-related genes TRAP, NFATc1 and c-Fos. Significance ORI may promote BMSCs differentiate into osteoblasts through the Wnt/β-catenin signaling pathway. At the same time, it may also inhibit the formation of osteoclasts mediated by RANKL. |
| Databáze: | OpenAIRE |
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