Melatonin rescues glucocorticoid-induced inhibition of osteoblast differentiation in MC3T3-E1 cells via the PI3K/AKT and BMP/Smad signalling pathways
Autor: | Lin Tao, Yihao Tian, Shui Qiu, Zheng Bo Tao, Lin Shen, Rui Zhao, Yue Zhu, Zunlei Gong |
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Rok vydání: | 2020 |
Předmět: |
Biomineralization
0301 basic medicine Bone Morphogenetic Protein 2 Smad Proteins SMAD 030226 pharmacology & pharmacy Dexamethasone General Biochemistry Genetics and Molecular Biology Cell Line Melatonin Mice Phosphatidylinositol 3-Kinases 03 medical and health sciences Calcification Physiologic 0302 clinical medicine Osteogenesis medicine Animals General Pharmacology Toxicology and Pharmaceutics Glucocorticoids Protein kinase B PI3K/AKT/mTOR pathway Osteoblasts Chemistry Cell Differentiation Osteoblast General Medicine Cell biology RUNX2 Blot 030104 developmental biology medicine.anatomical_structure Gene Expression Regulation Bone Morphogenetic Proteins Alkaline phosphatase Proto-Oncogene Proteins c-akt hormones hormone substitutes and hormone antagonists Signal Transduction medicine.drug |
Zdroj: | Life Sciences. 257:118044 |
ISSN: | 0024-3205 |
Popis: | Aims High-dose glucocorticoid (GC) administration causes osteoporosis. Many previous studies from our group and other groups have shown that melatonin participates in the regulation of osteoblast proliferation and differentiation, especially low concentrations of melatonin, which enhance osteoblast osteogenesis. However, the role of melatonin in glucocorticoid-induced osteoblast differentiation remains unknown. Materials and methods An examination of the expression of osteoblast differentiation markers (ALP, OCN, COLL-1), as well as alkaline phosphatase staining and alkaline phosphatase enzymatic activity assay to measure osteoblast differentiation and quantifying Alizarin red S staining to measure mineralization, were performed to determine the effects of dexamethasone (Dex) and melatonin on the differentiation of MC3T3-E1 cells. We used immunofluorescence staining to detect the expression of Runx2 in melatonin-treated MC3T3-E1 cells. The expression of mRNA was determined by qRT-PCR, and protein levels were measured by western blotting. Key findings In the present study, we found that 100 μM Dex significantly reduced osteoblast differentiation and mineralization in MC3T3-E1 cells and that 1 μM melatonin attenuated these inhibitory effects. We found that only inhibition of PI3K/AKT (MK2206) and BMP/Smad (LDN193189) signalling abolished melatonin-induced differentiation and mineralization. Meanwhile, MK2206 decreased the expression of P-AKT and P-Smad1/5/9 and LDN193189 decreased the expression of P-Smad1/5/9 but had no obvious effect on P-AKT expression in melatonin-treated and Dex-induced MC3T3-E1 cells. Significance These findings suggest that melatonin rescues Dex-induced inhibition of osteoblast differentiation in MC3T3-E1 cells via the PI3K/AKT and BMP/Smad signalling pathways and that PI3K/AKT signalling may be the upstream signal of BMP/Smad signalling. |
Databáze: | OpenAIRE |
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