BTK在粪肠球菌介导的炎症环境中的破骨作用

Role of BTK in osteoclasts stimulated by Enterococcus faecalis

目的在粪肠球菌脂磷壁酸(LTA)作用的炎症环境下,研究布鲁顿酪氨酸激酶(BTK)在破骨细胞中的作用,从而为根尖周炎的治疗提供实验依据。方法 PCR检测粪肠球菌LTA刺激破骨细胞后BTK基因水平的表达情况。以浓度300ng/mL的人重组蛋白BTK(recombinant human BTK,rhBTK)刺激破骨细胞,用CCK8法检测破骨细胞增殖和RT-PCR检测破骨细胞分化标志因子TRAP基因水平表达情况。结果破骨前体细胞5d诱导成功。PCR结果发现粪肠球菌LTA刺激后BTK和TRAP的mRNA表达量明显增高;免疫荧光可见LTA刺激后BTK在破骨细胞中的定位情况;300ng/mL rhBTK组可以促进破骨细胞增殖;PCR结果显示,加入rhBTK后,破骨细胞分化标志因子TRAP的mRNA水平升高。结论在粪肠球菌LTA作用的炎症环境下,BTK表达升高;增高BTK后,可以促进破骨细胞的增殖及分化。研究发现BTK参与了破骨细胞的炎症反应进程。

Objective To study the role of BTK in osteoclasts in the inflammatory environment stimulated by Enterococcus faecalis LTA,so as to provide experimental evidence for the treatment of periapical periodontitis.Methods PCR was used to detect the expression of BTK gene in osteoclasts stimulated by LTA.Osteoclasts were stimulated with human recombinant protein BTK at the concentration of 300 ng/mL.The proliferation of osteoclasts was detected by using CCK8 and the expression of TRAP gene was detected by using RT-PCR.Results RAW264.7 cells were successfully induced.The results of PCR showed that the expression of mRNA in BTK and TRAP significantly increased after stimulating by Enterococcus faecalis LTA.The localization of BTK in osteoclasts after LTA stimulation was seen by using immunofluorescence.300 ng/mL rhBTK promoted the proliferation of osteoclasts.PCR results showed that the level of mRNA of osteoclast differentiation marker TRAP increased after rhBTK was added.Conclusion The expression of BTK increased in the inflammatory environment stimulated by Enterococcus faecalis.Increased BTK can promote the proliferation and differentiation of osteoclasts.BTK was found involving in the inflammatory response of osteoclasts.