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Effect of Simvastatin on mRNA of Some Components of the Wnt Signaling Pathway in the Osteogenic Differentiation Process of Bone Marrow Stromal Cells of Rats

Author: ZhangLei
Tutor: ZhangLiu
School: North China Coal Medical
Course: Surgery
Keywords: Bone marrow stromal cells Osteoblasts RNA interference Wnt signaling pathway Simvastatin
CLC: R965
Type: Master's thesis
Year: 2009
Downloads: 87
Quote: 0
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Abstract


Purpose: To verify the simvastatin promote SD rat bone marrow stromal cells (BMSC) in vitro osteogenic differentiation on the basis of preliminary study the role of Wnt signaling in this process in order to further clarify its mechanism of action. Methods: female SD rats were, six weeks femur, tibia bone marrow cells, whole bone marrow culture method for primary and subculture. Were divided into four groups: control group (G1), the SIM group (G2), RNA interference group (G3), RNA interference SIM group (G4). Passage 2 after G3, G4 cells the the Axin2 mRNA interference plasmid transfection, four groups were changed after 48 hours with induction medium, while G2, G4 group 10-7mol / L SIM. 7 days after the administration of alkaline phosphatase (ALP) staining, 28 days after the von Kossa method to detect extracellular matrix mineralization; Application Real Time RT-PCR methods were used to detect administration of 7 and 14 days Axin2, b-catenin, osteocalcin (OCN), frizzled-2, LEF-1, Wnt5a mRNA expression. Results: the staining 1.ALP: G2 ALP expression was significantly higher than in G1, G3 and G4 group was no significant difference. 2.von Kossa staining: In addition to the G3 group, other groups were visible matrix mineralization Summary G2 matrix mineralization compared with other groups the most significant. Administration of 7 days real time RT-PCR detection: G2 group Axin2 mRNA expression was significantly lower than the G1 group the G2 group frizzled-2 mRNA expression was significantly higher than G1 group; G3 group of b-catenin and frizzled-2 mRNA was significantly lower than in G1, OCN, Lef-1, Wnt5a mRNA expression was significantly higher than in G1; G4 group of b-catenin mRNA expression was significantly higher than the G3 group, OCN, Lef-1, Wnt5a mRNA expression was significantly lower than G3 groups. (P lt; 0.05). 4 administered 14 days real time RT-PCR detection: G2 group Axin2, OCN, frizzled-2, LEF-1, Wnt5a mRNA expression were significantly higher than the G1 group; G3 group frizzled-2, LEF-1, Wnt5a mRNA was significantly higher than the G1 group; G3, G4 group OCN mRNA expression was significantly higher than, and frizzled-2, Lef-1 mRNA expression was significantly lower than the G3 group. (P lt; 0.05). Conclusion: Simvastatin promote BMSC differentiation into osteoblasts, accompanied by Wnt signaling pathway gene mRNA expression levels change. 2 Axin2 RNA interference, the Wnt signaling pathway activity increases, BMSC osteoblast differentiation late extracellular matrix mineralization diminished capacity. Axin2 RNA interference did not significantly affect the simvastatin and promote BMSC differentiation into osteoblasts role, simvastatin can be adjusted to changes in the level of expression of the RNA interference caused by the Wnt signaling pathway related factors Axin2, making it increasingly than normal.

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