牙髓炎症中白细胞介素-17调节单核细胞功能和分化的研究

Interleukin-17 Regulates the Function and Differentiation of Monocytes in Dental Pulp Inflammation

目的 探讨牙髓炎症中白细胞介素(IL)-17募集单核细胞并调节其分化的作用。方法 肿瘤坏死因子(TNF-α)刺激牙髓细胞,模拟炎症状态,实时荧光定量PCR技术(RT-qPCR)、酶联免疫吸附法(ELISA)检测IL-17的表达;IL-17重组蛋白刺激牙髓细胞,收集培养基上清,通过Transwell小室实验检测IL-17或受IL-17刺激的牙髓细胞培养基上清对单核细胞的趋化作用;采用结晶紫染色和吞噬实验分别检测IL-17或受IL-17刺激的牙髓细胞培养基上清对单核细胞贴壁和吞噬的影响;IL-17刺激人单核细胞系(THP-1)细胞,RT-qPCR和ELISA分别检测TNF-α、IL-1β的表达,流式细胞检测THP-1细胞表面标志物分子簇80(CD80)的表达变化。结果 牙髓炎症中IL-17表达上调,IL-17使单核细胞贴壁增多,吞噬功能增强,TNF-α、IL-1β及CD80表达上调。结论 IL-17在牙髓炎症中高表达,其本身对单核细胞有趋化作用,并可通过刺激牙髓细胞产生趋化因子2(CCL2)放大趋化作用;IL-17可以调控单核细胞向巨噬细胞分化。

Objective To explore the role of interleukin-17(IL-17)in recruiting monocytes and regulating the differentiation of monocytes during dental pulp inflammation.Methods Tumor necrosis factorα(TNF-α)stimulated human dental pulp cells cultured in vitro to induce inflammation.The expression of IL-17 was detected by real time quantitative polymerase chain reaction(RT-qPCR)and enzyme-linked immunosorbent assay(ELISA).IL-17 recombinant protein stimulated dental pulp cells and the supernatant was collected.Transwell assay was used to measure the chemotactic effect of IL-17 or the supernatant collected from dental pulp cells stimulated by IL-17 on monocytes.The crystal violet staining test and phagocytosis experiment were used to detect effect of IL-17 or the supernatant collected from dental pulp cells stimulated by IL-17 on the adhesion and phagocytosis of monocytes,respectively.IL-17 stimulated human monocyte cell line tohoku hospital pediatrics-1(THP-1)cells.RT-qPCR and ELISA were used to detect the expression of TNF-αand IL-1βof THP1 cells.Flow cytometry was used to detect the expression of the surface marker cluster of differentiation(CD80)of THP-1 cells.Results The expression of IL-17 was up-regulated in dental pulp inflammation.IL-17 promoted the adhesion and phagocytosis of monocytes,and up-regulated the expression of TNF-α,IL-1βand CD80.Conclusion IL-17 is up-regulated in inflamed dental pulp.IL-17 has a chemotactic effect on monocytes and can amplify the chemotactic effect by stimulating the production of chemokine 2(CCL2)in dental pulp cells.IL-17 can promote monocytes to differentiate into macrophages.