刺血疗法对急性痛风性关节炎大鼠局部IL-1β、IL-10启动子甲基化的影响

发布时间：2018-12-08 10:22

【摘要】：目的观察刺血疗法对急性痛风性关节炎大鼠造模关节局部白介素-1β(IL-1β)、白介素-10(IL-10)基因的相对表达及启动子区甲基化水平的影响,探讨刺血疗法治疗急性痛风性关节炎的相关表观遗传调控机制。方法 36只SD大鼠随机分为正常组、模型组、药物组、刺血组。右踝关节腔注射尿酸钠建立急性痛风性关节炎大鼠模型。刺血组在右侧昆仑穴点刺放血,药物组以布洛芬灌胃治疗。评价造模关节肿胀程度,光镜下观察关节腔组织形态学改变,qPCR和焦磷酸测序检测大鼠造模关节局部软骨及其周围软组织IL-1β、IL-10的mRNA表达水平及基因启动子区甲基化水平。结果与模型组、药物组比较,刺血组关节肿胀指数明显降低(P0.01)。刺血疗法能减少关节腔内尿酸盐结晶沉积,抑制炎性细胞的侵润,改善关节滑膜的组织形态结构。与模型组比较,刺血组IL-1βmRNA的相对表达量明显降低(P0.01)。但IL-1β基因启动子区甲基化水平与其mRNA表达不呈负相关。与正常组、模型组、药物组比较,刺血组IL-10 mRNA的相对表达量升高(P0.01),IL-10平均甲基化率降低(P0.05~0.01)。IL-10基因启动子甲基化水平与其mRNA表达呈显著负相关(r=-0.899,P0.01)。结论刺血疗法通过DNA甲基化的表观遗传修饰调控IL-10基因表达可能是刺血疗法发挥抗炎效应的重要机制之一。
[Abstract]:Objective to observe the effect of blood pricking therapy on the relative expression of interleukin-1 尾 (IL-1 尾) and interleukin-10 (IL-10) gene and the methylation level of promoter region in rat models of acute gouty arthritis. To explore the epigenetic regulation mechanism of blood pricking therapy for acute gouty arthritis. Methods 36 SD rats were randomly divided into normal group, model group, drug group and blood prick group. The rat model of acute gouty arthritis was established by injecting sodium uric acid into right ankle joint. The blood pricking group was punctured at right Kunlun point and the drug group was treated by intragastric administration of ibuprofen. To evaluate the degree of joint swelling, observe the morphologic changes of articular cavity under light microscope, and detect the IL-1 尾 of articular cartilage and its surrounding soft tissue in rats by qPCR and pyrosequencing. MRNA expression level and methylation level of gene promoter in IL-10. Results compared with model group and drug group, the joint swelling index of blood pricking group was significantly lower (P 0.01). Blood pricking therapy can reduce the deposition of uric acid crystal in articular cavity, inhibit the infiltration of inflammatory cells, and improve the morphology of synovium. Compared with the model group, the relative expression of IL-1 尾 mRNA was significantly decreased in the blood pricking group (P0.01). However, the methylation level of IL-1 尾 gene promoter was not negatively correlated with its mRNA expression. Compared with normal group, model group and drug group, the relative expression of IL-10 mRNA in blood pricking group increased (P0.01). The average methylation rate of IL-10 decreased (P0.05 / 0. 01), and the methylation level of IL-10 gene promoter was negatively correlated with its mRNA expression (r-0.899P0.01). Conclusion the regulation of IL-10 gene expression by epigenetic modification of DNA methylation in blood pricking therapy may be one of the important mechanisms of anti-inflammatory effect of blood pricking therapy.
【作者单位】：浙江医院推拿科;南京中医药大学第二临床医学院;湖南医药学院针灸教研室;第二军医大学长征医院南京分院针灸科;
【基金】：国家自然科学基金(81273840)
【分类号】：R245

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