沉默Periostin基因调节小鼠成骨细胞代谢对抑制破骨活性的作用

Silencing of Periostin gene modulates murine osteoblast metabolism to inhibit bone resorption activity

目的通过观察沉默Periostin后小鼠成骨细胞中特异性转录因子(Runx2)、核因子-κB受体活化因子配体(RANKL)及骨保护素(OPG)表达变化,探讨其影响破骨活性的分子机制。方法将小鼠成骨细胞株(MC3T3-E1)分为实验组和对照组,实验组使用LVpFU-GW-016PSC66473-1病毒转染,对照组用pFU-GW-016PSC53349-1病毒,保持两组细胞状态相当。用MDC法检测基因沉默效果,Western blot法检测两组细胞Runx2、RANKL和OPG蛋白表达情况。结果沉默Periostin后发现:实验组荧光表达较对照组明显增强(P<0.01);实验组Runx2和RANKL蛋白表达较对照组明显下调(P<0.05);OPG蛋白表达较对照组显著升高(P<0.05)。结论沉默Periostin可改变小鼠成骨细胞RANKL和OPG表达,并可能通过核因子-κB(NF-κB)信号通路介导影响Runx2基因,增强成骨细胞功能并抑制骨破坏。

Objective To investigate the molecular mechanisms by which silencing of the Periostin gene affects the expression of Runx2,receptor activator of nuclear factor-kappa B ligand(RANKL),and osteoprotegerin(OPG)in murine osteoblasts and its impact on bone resorption activity.Methods Murine osteoblast cell line MC3T3-E1 was divided into an experimental group and a control group.The experimental group was transfected with LVpFU-GW-016PSC66473-1 virus,while the control group received pFU-GW-016PSC53349-1 virus to ensure equivalent cellular conditions.The efficiency of gene silencing was assessed using MDC method,and the protein expression of Runx2,RANKL,and OPG in both groups was evaluated by Western blotting.Results Following Periostin silencing,the experimental group showed significantly enhanced fluorescence expression compared to the control group(P<0.01).Protein expression of Runx2 and RANKL was significantly decreased(P<0.05),whereas OPG expression was markedly increased(P<0.05)in the experimental group compared to the control group.Conclusion Silencing of Periostin alters RANKL and OPG expression in murine osteoblasts,potentially influencing the Runx2 gene via the nuclear factor-kappa B(NF-κB)signaling pathway.This enhancement of osteoblast function may contribute to the inhibition of bone resorption.