膝关节骨性关节炎患者IL-6水平变化以及NFAT1对IL-6作用机制研究

Change of IL-6 level and effect of NFAT1 on IL-6 in patients with knee osteoarthritis

目的检测不同病程膝关节骨性关节炎患者血液与滑膜液中白细胞介素-6(Interleukin-6,IL-6)含量,探讨其与活化T细胞核转录因子(Nuclear factor of activated T cell 1,NFAT1)表达水平在此类患者疾病进展中是否存在关联。方法选取自2017-05—2018-12住院治疗的56例膝关节骨性关节炎,轻度组8例,中度组31例,重度组17例。检测患者血液和滑膜组织中IL-6含量及NFAT1的mRNA表达水平,采用双荧光素酶报告基因验证NFAT1与IL-6启动子的相互作用。结果膝关节骨性关节炎患者滑膜液中IL-6的浓度明显高于血液,而外周血单个核细胞NFAT1的mRNA水平高于滑膜组织。重度组血液与滑膜液中IL-6含量高于轻度组和中度组,差异有统计学意义(P<0.05)。轻度组外周血单个核细胞与滑膜组织中NFAT1的mRNA水平低于中度组,而中度组低于重度组,差异有统计学意义(P<0.05)。在293T细胞中沉默NFAT1将显著降低野生型IL-6荧光报告质粒驱动的荧光素酶活性,但是突变型IL-6荧光报告质粒驱动的荧光素酶活性并无明显变化。结论IL-6与膝关节骨性关节炎的发生、发展相关,NFAT1作为一类具有广泛生理功能的转录因子参与介导膝关节骨性关节炎患者的IL-6表达,而IL-6高表达将加速疾病进展,因此在骨性关节炎早期即对NFAT1的作用进行抑制或许可以有效控制疾病进一步发展。

Objective To detect the content of interleukin-6(IL-6)in blood and synovial fluid of knee osteoarthritis with different courses,and investigate whether it is associated with nuclear factor of activated T cell 1(NFAT1)expression level in the progression of suc h kind of disease.Methods Fifty six cases of knee osteoarthritis were hospitalized from May 2017 to December 2018,8 cases in the mild group,31 cases in the moderate group and 17 cases in the severe group.IL-6 content and the NFAT1 mRNA expression levels in patient blood and synovial tissues were examined,and the double luciferase reporter was used to verify the interaction of NFAT1 with the IL-6 promoter using the luciferase gene.Results The concentration of IL-6 was significantly higher in the synovial fluid of knee osteoarthritis than that in the blood,while the mRNA levels of peripheral blood mononuclear cell NFAT1 were higher than that in the synovial tissue.IL-6 was higher in blood and synovial fluid than that of in mild group and moderate group,the differences were statistically significant(P<0.05).The mRNA levels of NFAT1 were lower in peripheral blood mononuclear cells and synovial tissues in the mild group and lower than in the moderate group,the differences were statistically significant(P<0.05).Silencing of NFAT1 in 293T cells would significantly reduce the luciferase activity driven by the wild-type IL-6 fluorescent reporter plasmid,but showed no significant change in the mutant IL-6 fluorescent reporter plasmid-driven luciferase activity.Conclusion IL-6 is associated with the occurrence and development of knee osteoarthritis.NFAT1 is involved as a class of transcription factors with a wide range of physiologic functions to mediate IL-6 expression in patients with knee osteoarthritis,while high IL-6 expression will accelerate disease progression,so inhibition of the effect of NFAT1 early in OA may effectively control the further disease development.