Application of galangin in prevention and treatment of osteoporosis by promoting osteogenic differentiation of bone marrow mesenchymal stem cells
A technology for bone marrow mesenchyme and osteoporosis, applied in the field of medicine, to achieve the effects of promoting osteogenic differentiation, promoting calcium salt nodule deposition, and increasing ALP activity
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Embodiment 1B
[0034] Determination of ALP activity in the process of embodiment 1BMSCs osteogenic differentiation
[0035] Alkaline phosphatase (ALP) is a marker for the maturation, differentiation and mineralization of early osteoblasts, and provides a material basis for the mineralization of bone cells to form bones. In this example, ALP activity was used as an index to evaluate the effect of galangin on the early osteogenic differentiation of BMSCs.
[0036] (1) Experimental grouping
[0037] This embodiment is divided into four groups altogether, i.e. control group (only use osteogenesis induction medium to carry out induction culture), drug low dosage group (use osteogenesis induction medium to dilute to the concentration of galangin liquid of 5 μ M), drug Middle dose group (galangin solution diluted to a concentration of 10 μM with an osteogenic induction medium), and high dose group (galangin solution diluted to a concentration of 20 μM with an osteogenic induction medium).
[0038] ...
Embodiment 2B
[0042] Alizarin red staining of embodiment 2 BMSCs osteogenic differentiation calcium salt nodules
[0043] Alizarin red staining is generally used to identify the mineralization of osteoblasts cultured in vitro. In this example, this staining method was used to evaluate the effect of galangin on the osteogenic differentiation of BMSCs.
[0044] (1) Measurement method
[0045] The resuscitated and digested bone marrow mesenchymal stem cells were administered according to the grouping and method in Example 3. After 14 days of osteogenic differentiation, the cells were observed under a microscope, and calcium salt nodules formed by osteogenic differentiation were deposited on the cell surface. After the cells were fixed, the cells were washed with PBS for 3 times, and then stained with Alizarin Red. After incubation at room temperature for 15 minutes, the cells were slowly washed with PBS for 3 times. The formation of calcium salt nodules was observed under a microscope, and the...
Embodiment 3
[0048] Example 3 Detection of Osteogenic Differentiation Levels by Western Blotting
[0049] (1) Determination method: The recovered and digested bone marrow mesenchymal stem cells were also administered according to the grouping and method in Example 3. After 14 days of osteogenic differentiation, cell samples were collected by adding cell lysate, and Western blotting was performed. The expression levels of the indicator proteins Runx2, OCN, and OPN proteins (with GAPDH as the internal reference) in the process of osteogenic differentiation were analyzed by using the method (references for specific operation methods: Tao K, Xiao D, Weng J, et al. Berberine promotes bonemarrow-derived mesenchymal stem cells osteogenic differentiation via canonical Wnt / β-catenin signaling pathway[J].Toxicology Letters,2016:68-80.).
[0050] (2) Measurement results: image 3 The assay results shown showed that the expression levels of Runx2, OCN, and OPN proteins in the cells were significantly...
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