炎症因子白介素-6对骨细胞表达RANKL的影响

IL-6 Enhances the Expreesion of RANKL in Osteocyte

目的 :通过研究炎症因子白介素-6对骨细胞表达核因子-κB受体活化因子配基(RANKL)的影响,为更好地理解炎症因子在骨细胞参与骨重塑中的作用,提供理论基础。方法:体外培养MLO-Y4细胞系细胞,并应用不同浓度的人工合成白介素-6(IL-6)和白介素-6受体(IL-6R),共同作用刺激骨细胞24 h后,采用实时荧光定量PCR、免疫印迹试验、免疫荧光等方法,检测RANKL以及酪氨酸激酶2(JAK2)的磷酸化蛋白的表达变化。结果:应用不同浓度人工合成的IL-6和IL-6R,共同作用刺激骨细胞后,RANKL的表达水平随着IL-6的浓度增加而增加,并且IL-6通路中的JAK2蛋白磷酸化亦增加,提示IL-6及其受体可以促进骨细胞表达RANKL,并且与JAK2的活化密切相关。结论 :炎症因子IL-6可以通过增加JAK2蛋白的磷酸化,来促进骨细胞表达RANKL。

Objective： To study the effects of IL-6 on the expression of RANKL in osteocytes. Methods： MLO-Y4 cells were pre-treated with 10, 30, 50 or 100 ng/mL IL-6/IL-6R for 24 hours. The effect of IL-6/IL-6R on RANKL and p-JAK2 mRNA and protein expression in MLO-Y4 cells was detected by real-time PCR, Western-blot, and immunofluorescence. Results： After treating MLO-Y4 cells with 10, 30, 50, 100 ng/mL of IL-6/IL-6R, expression intensities of RANKL mRNA and protein increased in a dose-dependent manner. In addition, the increased expression of RANKL after treatment with IL-6/IL-6R was also shown by immunofluorescence. Additionally, p-JAK2 protein was upregulated after treatment with IL-6/IL-6R. Conclusion： IL-6 enhances expression of RANKL and JAK2 activation in the osteocyte-like cell line MLO-Y4. This observation suggests a relationship may be existed between RANKL expression and p-JAK2.