Zip1 转染骨髓间充质干细胞对 Wnt1/β-catenin信号成骨通路的影响

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目的 观察ZIP1 siRNA转染的骨髓间充质干细胞对Wnt1/β-catenin信号成骨通路的影响.方法 从成年雄性新西兰兔股骨骨髓中提取骨髓间充质干细胞并培养传代,然后使用0. 03 mol/L,0. 09 mol/L,0. 15 mol/L,0. 21 mol/L酒精进行培养,经ZIP1 siRNA及ZIP1表达转染后,检测各组骨髓间充质干细胞中ALP、I型胶原、OCN及Runx2蛋白的表达量.结果 与空白对照组相比,不同酒精组的ALP、I型胶原、OCN及Runx2 蛋白表达量均有所下降,随着浓度升高,其表达量逐渐下降,但0. 03 mol/L组中ALP及OCN下降不明显,统计学无明显意义(P>0. 05),其他各组均有明显的统计学意义(P<0. 05),不同酒精组中的I型胶原及Runx2与正常组之间差异均有明显统计学意义(P<0. 05).与空白对照组对比,ZIP1表达组ALP、I型胶原及OCN表达量明显升高,差异具有明显的统计学意义(P<0. 05),ZIP1siRNA组ALP、OCN表达量明显降低,差异具有明显的统计学意义(P<0. 05),而I型胶原表达量下降不明显,差异无统计学意义(P>0. 05);与ZIP1siRNA组相比,ZIP1表达组ALP、I型胶原及OCN表达量明显升高,差异有统计学意义(P<0. 05).结论 ZIP1能促使骨髓间质干细胞向成骨分化,从而促进骨修复及骨代谢.沉默ZIP1表达能抑制骨髓间充质干细胞的WNT1/β-catenin信号成骨通路的表达,从而抑制促进细胞的成骨分化能力.“,”Objective To study the effect of BMSCs transfected by ZIP1 ( Zrt/Irt-like Protein 1) on Wnt1/β-catenin signaling pathway in the process of osteogenic differentiation. Methods BMSCs were extracted from bone marrow of the femurs of adult male New Zealand rabbits and cultured under different concentrations of alcohol (0. 03 mol/L, 0. 09 mol/L, 0. 15 mol/L, and 0. 21 mol/L, respectively). Cells were transfected by ZIP1 siRNA and ZIP1. The expression ALP, collagen type I, OCN, and Runx2 were detected in BMSCs. Results Compared with the control group, the protein expression of ALP, collagen type I, OCN, and Runx2 decreased in different alcohol groups. With the increase of alcohol concentration, the expression decreased gradually. The decrease of ALP and OCN was not significant in 0. 03 mol/L group (P >0. 05). The difference was statistically significant (P<0. 05) in other groups. The differences of the expression of collagen type I and Runx2 between different alcohol groups and normal group were statistically significant (P<0. 05). Compared with the control group, the expression of ALP, collagen type I and OCN increased significantly in the ZIP1 group, and the difference was significant (P<0. 05). The expression of ALP and OCN, except the type I collagen, decreased significantly in the ZIP1 siRNA group, and the difference was significant (P<0. 05). Compared with ZIP1 siRNA group, the expression of ALP, collagen type I, and OCN increased significantly in the ZIP1 group, and the difference was statistically significant (P<0. 05). Conclusion ZIP1 promotes the osteogenic differentiation of BMSCs and promotes bone repair and bone metabolism. ZIP1 siRNA suppresses the osteogenic differentiation of BMSCs by activating Wnt1/β-catenin signaling pathway.
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