■探讨组蛋白H3K27me3甲基化及其调控酶JMJD3和EZH2在强直性脊柱炎(AS)Th17细胞分化中的作用,揭示它们在AS发病机制中的潜在参与,分析H3K27me3的甲基化状态及其与Th17相关因子的相互作用,为AS的临床治疗提供新的策略和靶点。
■本研究共纳入84例AS患者(42例活动性AS患者和42例AS稳定期患者),84名健康志愿者作为对照。收集血液样品。分离外周血单核细胞。进行ELISA测定以检查Th17细胞和相关细胞因子IL-21、IL-22和IL-17。RORc的mRNA表达,通过RT-PCR分析JAK2和STAT3,RORc的蛋白质表达,JAK2/STAT3通路蛋白,通过Western印迹测定H3K27me3和相关蛋白酶(EZH2和JMJD3)。采用Pearson相关性分析H3K27me3、EZH2和JMJD3与Th细胞分化关键信号通路分子的相关性。
■RORc的mRNA表达,活动期组JAK2、STAT3明显高于稳定期组(P<0.05)。活性相组H3K27me3和EZH2的相对灰度值低于稳定相组,低于对照组,差异有统计学意义(P<0.05)。JMJD3、RORc、活动相组的JAK2、pJAK2、STAT3、pSTAT3蛋白明显高于稳定期组,均高于对照组(均P<0.05)。活动期组Th17比例及炎症因子表达水平高于其他两组(P<0.05)。H3K27me3与RORc呈负相关,JAK2、STAT3、IL-17、JMJD3与JAK2、STAT3、IL-17呈正相关,EZH2与JAK2、STAT3、IL-17呈负相关(均P<0.05)。
■H3K27me3在AS中的低表达受基因位点JMJD3和EZH2的影响,可以调节Th17细胞的分化,从而在AS的发病和进展中发挥作用。
UNASSIGNED: To investigate the roles of histone H3K27me3 methylation and its regulatory enzymes JMJD3 and EZH2 in the differentiation of Th17 cells in ankylosing spondylitis (AS), to unveil their potential involvement in the pathogenesis of AS, and to provide new strategies and targets for the clinical treatment of AS by analyzing the methylation state of H3K27me3 and its interactions with Th17-related factors.
UNASSIGNED: A total of 84 AS patients (42 active AS patiens and 42 patients in the stable phase of AS) were enrolled for the study, while 84 healthy volunteers were enrolled as the controls. Blood samples were collected. Peripheral blood mononuclear cells were isolated. ELISA assay was performed to examine Th17 cells and the relevant cytokines IL-21, IL-22, and IL-17. The mRNA expressions of RORc, JAK2, and STAT3 were analyzed by RT-PCR, the protein expressions of RORc, JAK2/STAT3 pathway protein, H3K27me3 and the relevant protease (EZH2 and JMJD3) were determined by Western blot. Correlation between H3K27me3, EZH2 and JMJD3 and the key signaling pathway molecules of Th cell differentiation was analyzed by Pearson correlation analysis.
UNASSIGNED: The mRNA expressions of RORc, JAK2, and STAT3 were significantly higher in the active phase group than those in the stable phase group ( P<0.05). The relative grayscale values of H3K27me3 and EZH2 in the active phase group were lower than those of the stable phase group, which were lower than those of the control group, with the differences being statistically significant ( P<0.05). The relative grayscale values of JMJD3, RORc, JAK2, pJAK2, STAT3, and pSTAT3 proteins were significantly higher in the active phase group than those in the stable phase group, which were higher than those in the control group (all P<0.05). The proportion of Th17 and the expression level of inflammatory factors in the active period group were higher than those in the other two groups (P<0.05). H3K27me3 was negatively correlated with RORc, JAK2, STAT3, and IL-17, JMJD3 was positvely correlated with JAK2, STAT3, and IL-17, and EZH2 was negatively correlated with JAK2, STAT3, and IL-17 (all P<0.05).
UNASSIGNED: The low expression of H3K27me3 in AS is influenced by the gene loci JMJD3 and EZH2, which can regulate the differentiation of Th17 cells and thus play a role in the pathogenesis and progression of AS.