抑制p38MAPK信号转导通路对钛颗粒诱导RAW264.7巨噬细胞的影响

The Effect of Inhibition of p38MAPK Signaling Transduction Passway on Titanium Particles-induced RAW264.7 Macrophages

目的探讨抑制p38MAPK信号转导通路对钛(Ti)颗粒诱导RAW264.7巨噬细胞的影响及可能机制。方法体外培养小鼠RAW264.7巨噬细胞并通过加入不同剂量的p38MAPK信号转导通路抑制剂SB203580,分为对照组、SB20358060 nmol·L^(-1)组、SB203580120 nmol·L^(-1)组、SB203580240 nmol·L^(-1)组;TRAP法检测各组小鼠RAW264.7巨噬细胞转化为破骨细胞的能力;MTT检测各组小鼠RAW264.7巨噬细胞的增殖;ELISA法检测细胞TNF-α和MMP-9含量的表达;实时荧光定量RT-PCR检测SB203580对Ti颗粒刺激RAW264.7巨噬细胞分泌TNF-α、MMP-9 mRNA的表达。结果TRAP染色结果显示,与对照组比较,抑制剂SB203580能够明显抑制小鼠钛颗粒诱导下RAW264.7巨噬细胞向破骨细胞的增殖、成熟和活化。加入SB203580后,Ti颗粒刺激的RAW264.7巨噬细胞分泌的炎性因子TNF-α和MMP-9的蛋白表达降低(P均<0.01);抑制剂SB203580使TNF-α和MMP-9的mRNA的表达量下调(P均<0.01)。结论抑制p38MAPK信号转导通路能够抑制Ti颗粒刺激小鼠RAW264.7巨噬细胞向破骨细胞转化,从而进一步抑制破骨细胞的增殖、分化和成熟,最终抑制骨溶解。

Objective To investigate the effect and mechanism of inhibition of p38MAPK signal transduction pathway on titanium particles-induced RAW264.7 macrophages.Methods RAW264.7 macrophages were cultured in vitro and divided into control group,SB20358060 nmol·L^(-1) group,SB203580120 nmol·L^(-1) group,SB203580240 nmol·L^(-1) group.After the p38MAPK signal transduction pathway inhibitor SB203580 was added at different doses to inhibit the p38MAPK signal transduction pathway,the effect on the ability of RAW264.7 macrophages to transform into osteoclasts in each group was detected by the TRAP method;MTT was used to detect the proliferation of RAW264.7 macrophages in each group of mice;TNF-αand MMP-9 levels were detected by ELISA;real-time fluorescent quantitative RT-PCR was used to detect SB203580 on titanium particles to stimulate RAW264.7 macrophages to secrete mRNA expression of TNF-αand MMP-9.Results Compared with the control group,inhibition of p38MAPK signal transduction pathway could significantly inhibit the proliferation of RAW264.7 macrophages,and the protein expression of inflammatory factors TNF-αand MMP-9.Inhibition of the p38MAPK signal transduction pathway could inhibit the mRNA expression of TNF-αand MMP-9 significantly(P all<0.01).Conclusion Inhibition of p38MAPK signal transduction pathway can significantly inhibit titanium particles from stimulating the transformation of RAW264.7 macrophages to osteoclasts,thereby further inhibiting the proliferation,differentiation and maturation of osteoclasts,and thus inhibiting their osteolysis.