陆军军医大学学报 (Apr 2023)
Effect and mechanism of magnesium on osteogenic differentiation in porcine aortic valve interstitial cells
Abstract
Objective To explore the effect of magnesium ion on osteogenic differentiation of porcine aortic valve interstitial cells (AVICs) and its potential mechanism. Methods Porcine AVICs were isolated from fresh pig aortic valve samples through two-step digestion combined with type Ⅱ collagen, and then cultured and identified with immunofluorescence staining and flow cytometry for cell phenotypes. The cells were randomly divided into blank control group (Blank Group), osteogenic induction medium group (OIM Group) and magnesium treatment group (1, 2 and 3 mmol/L MgSO4 solution for 7 d). The expression of osteogenic differentiation markers, osteopontin (OPN) and Runt-related transcription factor 2 (RUNX2) were detected by Western blotting. Alizarin red staining was adopted to observe calcium deposition in the cells of each group. Subsequently, the changes in PI3K/Akt and MAPK/ERK1/2 signaling pathways were measured by Western blotting. Results Our obtained porcine AVICs had high expression of Vimentin, low expression of α-SMA, and no expression of CD31, with an ideal purity of porcine AVICs for subsequent experiments. Western blotting showed that the expression of osteogenic markers OPN and RUNX2 was significantly higher in the OIM Group than the Blank Group (P < 0.01), and these effects induced by OIM were obviously inhibited with 3 mmol/L magnesium treatment (P < 0.01). Alizarin red staining displayed that calcium salt deposition was remarkably increased in the OIM Group than the Blank Group, and this increment was inhibited after magnesium treatment. The results of Western blotting indicated that magnesium treatment resulted in decreased phosphorylation of PI3K/AKT and MAPK/ERK1/2 induced by OIM (P < 0.01). Conclusion Magnesium may partially inhibit the osteogenic differentiation of porcine AVICs induced by OIM in vitro through PI3K/AKT and MAPK/ERK1/2 signaling pathways.
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