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Objective To study the effect of miR-124-3p targeting Axinl 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 Axin 1 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- l 24-3p and Axin 1. 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 result of Alizarin red staining and ALP staining were detected. The expression of Mir-124-3p and Runx2 mRNA, Alizarin red staining, ALP staining result 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 HbA 1 C were up-regulated, [3-CTX and SOST were up-regulated, and OC and BMD were downregulated 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 result showed that Axinl 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- l 24-3p and Axin 1 could be targeted binding. The expression of Mir- l 24-3p 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 Axin 1. [ABSTRACT FROM AUTHOR]

Objective The correlation between related pathways and postmenopausal osteoporotic fractures (PMOPF) is analyzed by detecting the expressions of LRP5, ß/catenin?,Runx2, C-myc, Osterix, OPG, RANKL, and LGR4 in the serum and bone tissues in PMOPF patients and in patients with non-osteoporosis fractures after menopause (control group). Methods The patients with tibial fractures were divided into control group and PMOPF group. Total RNA of the bone tissue was extracted using RNAiso Plus method . RT-qPCR method was used to detect the expression of each factor. The levels of serum factors were detected with ELISA method in the control group. PMOPF group was divided into A-F group according to the blood collection time interval. The changes between the control group and the PMOPF group and between the adjacent groups in the PMOPF group were compared. Results (1) The expressions of LRP5, ß-catenin, Runx2, C-myc, Osterix, OPG, and LGR4 in PMOPF group were lower than those in control group (P<0.05) detected by RT-qPCR. The expression of RANKL increased significantly (P<0. 05). ( 2) The serum levels of LRP5, ß-catenin, Runx2, C-myc, Osterix, OPG, and LGR4 decreased significantly ( P< 0.05) and RANKL increased significantly (P<0-05) in the A-F group of PMOPF group compared with those in the control group detected with ELISA method . LRP5 and Runx2 were the lowest in Group B. ß-catenin and C-myc were the lowest in Group C. RANKL was the highest in Group C. Osterix was the lowest in Group D. OPG and LGR4 were the lowest in Group E. Conclusion The related factors in Wnt / ß-catenin, BMP-2/ Runx2 / Osterix, OPG/ RANKL, LGR4 / RANKL pathways are closely related to the occurrence of PMOPF. LRP5 and Runx2 decreased to the lowest level within 3 days after fracture. ß-catenin and C-myc decreased to the lowest level within 7 days after fracture. The result show that the changes in osteogenesis are consistent. Osterix decreased to the lowest level within 14 days after fracture. OPG and LGR4 decreased to the lowest level within 28 days after fracture which might be related to the difficulty of short-term healing of PMOPF. RANKL increased to the highest level within 7 days after fracture which might be associated with the increase in bone formation after PMOPF. [ABSTRACT FROM AUTHOR]