| Objective To study the effect of miR-124-3p targeting Axin1 on osteogenic differentiation of bone marrow mesenchymal stem cells (BMSC) in diabetic osteoporosis rats. Methods Twenty diabetic rat model-SPF-grade SD female rats were divided into control group (n = 10) and experimental group (n = 10). After drug injection, blood biochemical indexes and bone mineral density were detected. BMSCs were cultured and identified with flow cytometry. The expression of miR-124-3p mRNA in control group and experimental group was detected. Mir-124-3p (mimics) was over-expressed for 7, 14, and 21 days. The osteogenic ability of ALP and Alizarin red staining was detected. On day 7, Mir-124-3p mRNA expression level was detected. Cells in the experimental group were transfected with Mir-124-3p mimics and divided into high glucose group and low glucose group, and Axin1 mRNA level was detected. The vector was constructed and co-transfected with mimics. The luciferase reporter gene was used to detect the targeted binding of Mir-124-3p and Axin1. BMSCs in the experimental group were divided into high glucose group and low glucose group, and the expression of Mir-124-3p and Runx2 mRNA and the results of Alizarin red staining and ALP staining were detected. The expression of Mir-124-3p and Runx2 mRNA, Alizarin red staining, ALP staining results in the experimental group mimics in high glucose group and low glucose group were detected. Results Serum biochemical indexes were detected with ELISA. TG, TC, FPG, and HbA1C were up-regulated, β-CTX and SOST were up-regulated, and OC and BMD were down-regulated in the experimental group. The expressions of CD73 and CD105 were positive while CD34 and CD45 were negative. These matched the antigenic characteristics of mesenchymal stem cells and were confirmed as BMSCs. The expression of Mir-124-3p in experimental group was significantly lower than that in control group. Mir-124-3p (mimics) was over-expressed on days 7, 14, and 21, and ALP and Alizarin red staining increased gradually with time. On day 7, the expression level of Mir-124-3p increased significantly. Cells in the experimental group were transfected with Mir-124-3p mimics. The results showed that Axin1 mRNA expression level was up-regulated in the high glucose group, but decreased in the transfection with Mir-124-3p mimics. Luciferase reporter gene detection of Mir-124-3p and Axin1 could be targeted binding. The expression of Mir-124-3p and Runx2 decreased significantly by high glucose intake of BMSCs in the experimental group by. High glucose inhibited osteogenic differentiation and reduced calcium deposition and ALP expression. The expression of Runx2 in Mir-124-3p over-expression (mimics) under hyperglycemia in high glucose + control group was significantly lower than low glucose + control group. Mir-124-3p mimics + high glucose were significantly higher than those of high glucose + control group. Alizarin red staining and ALP staining in high glucose + control group was significantly lower than in low glucose + control group. Mir-124-3p mimics + high glucose was significantly higher than those of high glucose + control group. Conclusion miR-124-3p promotes osteogenesis of BMSC in rats with diabetic osteoporosis by targeted inhibition of Axin1. |