Epimedium-Derived Flavonoids Modulate the Balance between Osteogenic Differentiation and Adipogenic

来源 :Chinese Journal of Integrative Medicine | 被引量 : 0次 | 上传用户:qingmeizhujiulyx
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Objective:To observe the function of wnt/β-catenin signal pathway on the process that epimedium-derived flavonoids(EFs) regulate the balance between osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats,and to provide an experimental evidence for the mechanism of EFs on treating postmenopausal osteoporosis.Methods:Bone marrow stromal cells from ovariectomized rats were separated and cultivated in the condition of osteoinductive medium or liquid medium for 15 days.Low-(1μg/mL),medium-(10μg/mL) and high-(100μg/mL) dose EFs were administrated correspondingly.Alkaline phosphatase(ALP) staining,ALP activity determination,oil red O staining and realtime polymerese chain reaction(RT-PCR) were used to determine the effect of EFs on osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats.Moreover,in order to explore the mechanism of EFs on osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats,Dickkopf-related protein 1(DKK1) was used in the medium group.Enzymelinked immunosorbent assay(ELISA) and RT-PCR were used to determine mRNA levels ofβ-catenin,low density lipoprotein receptor-related protein 5(LRP5) and T cell factor(TCF) protein,known as wnt/β-catenin signal pathway related factors.Results:EFs increased mRNA expression levels of ALP and early osteoblast differentiation factors,such as runt-related transcription factor 2(Runx2),osteocalcin and collagenⅠ,and decreased mRNA expression levels of fat generation factors,such as peroxisome proliferator activated receptor gamma 2(PPARγ-2) and CCAAT enhancer-binding protein-α(C/EBPα) in a dose-dependent manner.While osteoblast differentiation factors were down-regulated,fat generation factors were up-regulated when DKK1 was applied.Also EFs up-regulated mRNA expression levels ofβ-catenin,LRP5 and TCF protein which could be blocked by DKK1.Conclusion:EFs regulate the balance between osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats by activating wnt/β-catenin signal pathway, which may be an important molecular mechanism of EFs on treating postmenopausal osteoporosis. Objective: To observe the function of wnt / β-catenin signaling pathway on the process that epimedium-derived flavonoids (EFs) regulate the balance between osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats, and to provide an experimental evidence for the mechanism of EFs on treating postmenopausal osteoporosis. Methods: Bone marrow stromal cells from ovariectomized rats were separated and cultivated in the condition of osteoinductive medium or liquid medium for 15 days. Low- (1 μg / mL), medium- (10 μg / mL) ALP activity determination, oil red O staining and realtime polymerese chain reaction (RT-PCR) were used to determine the effect of EFs on osteogenic differentiation, and high- (100 μg / mL) dose EFs were administrated correspondingly.Alkaline phosphatase and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats. More over, in order to explore the mechanism of EFs on osteogenic differentiation and adipogenic d In differentiation of bone marrow stromal cells of ovariectomized rats, Dickkopf-related protein 1 (DKK1) was used in the medium group. Enzymelinked immunosorbent assay (ELISA) and RT-PCR were used to determine mRNA levels of β- catenin, low density lipoprotein receptor- Related proteins 5 (LRP5) and T cell factor (TCF) protein, known as wnt / β-catenin signal pathway related factors. Results: EFs increased mRNA expression levels of ALP and early osteoblast differentiation factors, such as runt-related transcription factor 2 (Runx2), osteocalcin and collagen I, and decreased decreased mRNA expression levels of fat generation factors, such as peroxisome proliferator activated receptor gamma 2 (PPARγ-2) and CCAAT enhancer-binding protein-α (C / EBPα) .While osteoblast differentiation factors were down-regulated, fat generation factors were up-regulated when DKK1 was applied. Als EFs up-regulated mRNA expression levels of β- catenin, LRP5 and TCF protein which could be blocked by DKK1.Conclusion: EF s regulate the balance between osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats by activating wnt / β-catenin signaling pathway, which may be an important molecular mechanism of EFs on treating postmenopausal osteoporosis.
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