基于microRNA调控的针刺抑制慢性疲劳综合征免疫炎性反应的机制研究

Chronic fatigue syndrome (CFS) is considered as one of the public health problems affecting human health in 21st Century. It’s urgently calling for a disease biomarker of high sensitivity and specificity as an indicators for disease diagnosis and treatment evaluation. The miRNAs may play a key role in pathological disorders of CFS, and may become the potential biomarker of diagnose, pathogen and curative effect for it. Acupuncture is effective in treating CFS, but the adjustment mechanism of the mechanism of CFS is still to be further deepened. Therefore, the project focus on CFS subjects, use Solexa sequencing technology to screen CFS circulating miRNA expression spectrum, and analyze potential biomarkers of CFS. At the same time, centring on the hypothesis that "acupuncture inhibit the immunological.inflammatory reaction of CFS by influencing miRNA-X expression through regulating NF-κB, P53 signal pathway”, we will conduct a research on CFS animal model, use RT-qPCR and liquid chip technology, find out whether acupuncture inhibit inflammation of CFS by influencing the expression of CFS circular miRNA through regulating NF-κB, P53 signal pathway, and reveal the efficacious mechanism to enrich the scientific connotation of acupuncture for CFS.

慢性疲劳综合征（CFS）被认为是21 世纪影响人类健康的公共卫生问题之一，目前亟须灵敏度和特异性相对较高的疾病标记物作为疾病确诊和疗效呈现的指标。miRNA可能在CFS 各种病理失调中发挥关键作用，可能成为CFS 诊断、发病机制、疗效的潜在生物标记物。针灸治疗CFS 疗效肯定，但对CFS 各病理机制间的调整机制还有待深入。因此，本项目以CFS受试者为研究对象，采用Solexa 测序技术筛选CFS 循环miRNA 表达谱，解析CFS 潜在的生物标记物。同时围绕“针刺通过影响miRNA-X表达调控NF-κB、P53信号通路，抑制CFS免疫炎性反应而发挥治疗作用”的工作假说，以CFS动物模型为研究载体，运用RT-qPCR、液相芯片技术等，明确针刺通过影响CFS载体miR-X 的表达调控NF-κB、P53 信号通路抑制免疫炎性反应，实现临床疗效的作用机制，以丰富针灸治疗CFS的科学内涵。

本项目采用高通量测序技术和RT-qPCR技术，发现与30例健康受试者相比较，30例符合CDC1994年和2003年诊断标准的中国慢性疲劳综合征患者，有84个显著变化的miRNA，主要富集于调节免疫炎症、氧化与硝化应激和能量代谢等相关信号通路；采用临床试验和动物实验，初步发现miR-184显著降低，可能与CFS患者免疫力低下（NK细胞杀伤活性降低）和认知功能障碍（工作记忆及信息加工能力损害）有关；初步证实，针刺改善CFS患者和CFS模型大鼠NK细胞杀伤活性降低，可能通过miR-184/ NFAT2/NKCC和miR-34a-5p/ERK1/2/NKCC信号通路实现调控；针刺改善CFS患者和CFS模型大鼠工作记忆及信息加工能力损害，可能通过miR-184/5-HT/Gnai/PLC/Casp-3信号通路机制。