Wuhan hospital of traditional Chinese Medicine
Hubei Provincial Health Commission Project
目的：本实验研究穴位埋线通过调控PD-1/OX40信号通路抗类风湿关节炎的作用及其机制。方法：取25只SD雌性大鼠，用完全弗氏佐剂进行诱导，建立类风湿关节炎模型大鼠，然后进行穴位埋线和药物治疗，将大鼠分成5组：对照组、模型组、来氟米特组、穴位埋线组和穴位埋线 来氟米特组，通过评价关节炎指数判断炎症程度；ELISA检测血清IL-6和IL-8的含量；HE染色观察滑膜炎性细胞浸润、滑腹组织、纤维组织、巨噬细胞増生及血管形成等情况；流式检测PD-1/OX40及CD4 CD28-PD-1lowOX40 T细胞亚群含量。结果：除对照组外，各组大鼠踝关节至跖骨处或掌关节出现红斑和中度肿胀，在14天以后模型组评分没有明显的变化，而来氟米特组、穴位埋线组和穴位埋线 来氟米特组炎症因子评分开始下降，其中穴位埋线 来氟米特组下降最快。模型组血清中IL-6、IL-8的含量显著升高（P<0.01），氟米特组、穴位埋线组和穴位埋线 来氟米特组大鼠血清中IL-6、IL-8的含量显著下降（P<0.01）。模型组大鼠滑膜组织遭到破坏，有大量的炎症细胞浸润，血管扩张，纤维细胞增生，在进行来氟米特和穴位埋线处理后大鼠滑膜组织中炎症细胞浸润明显降低，血管增生被抑制，且在来氟米特和穴位埋线共同作用时效果最明显。模型组中大鼠的PD-1lowOX40 的含量显著降低（P<0.01），CD4 CD28-PD-1lowOX40 T细胞含量显著升高（P<0.01），穴位埋线组和来氟米特 穴位埋线组的PD-1lowOX40 的含量显著增加（P<0.01），氟米特组、穴位埋线组和穴位埋线 来氟米特组大鼠的CD4 CD28-PD-1lowOX40 T细胞含量显著下降（P<0.01）。结论：穴位埋线能够改善类风湿关节炎症状，并且与来氟米特具有联合治疗的作用，其作用机制可能与PD-1/OX40信号通路有关。
Objective: This experiment studied the effect and mechanism of acupoint catgut embedding on rheumatoid arthritis by regulating PD-1 / OX40 signal pathway. Methods: 25 female SD rats were induced with complete Freund's adjuvant to establish rheumatoid arthritis model rats, and then treated with acupoint catgut embedding and drug therapy. The rats were divided into 5 groups: control group, model group, leflunomide group, acupoint catgut embedding group and acupoint catgut embedding leflunomide group. The degree of inflammation was judged by evaluating the arthritis index; The contents of serum IL-6 and IL-8 were detected by ELISA; He staining was used to observe synovitis cell infiltration, smooth abdominal tissue, fibrous tissue, macrophage proliferation and angiogenesis; The contents of PD-1 / OX40 and CD4 cd28-pd-1lowox40 T cell subsets were detected by flow cytometry. Results: except the control group, there were erythema and moderate swelling from ankle to metatarsal or metacarpal joint in each group. After 14 days, there was no significant change in the score of the model group, but the inflammatory factor scores of leflunomide group, acupoint catgut embedding group and acupoint catgut embedding leflunomide group began to decline, and the acupoint catgut embedding leflunomide group decreased the fastest. The contents of IL-6 and IL-8 in serum of model group increased significantly (P < 0.01), and the contents of IL-6 and IL-8 in serum of rats in flutamide group, acupoint catgut embedding group and acupoint catgut embedding leflunomide group decreased significantly (P < 0.01). The synovial tissue of the model group was damaged, with a large number of inflammatory cell infiltration, vascular dilatation and fibroblast proliferation. After leflunomide and acupoint catgut embedding treatment, the inflammatory cell infiltration in the synovial tissue of the model group was significantly reduced and the vascular proliferation was inhibited, and the effect was the most obvious when Leflunomide and acupoint catgut embedding worked together. In the model group, the content of pd-1lowox40 decreased significantly (P < 0.01), the content of CD4 cd28-pd-1lowox40 T cells increased significantly (P < 0.01), and the content of pd-1lowox40 in acupoint catgut embedding group and leflunomide acupoint catgut embedding group increased significantly (P < 0.01) The content of CD4 cd28-pd-1lowox40 T cells in acupoint catgut embedding group and acupoint catgut embedding leflunomide group decreased significantly (P < 0.01). Conclusion: acupoint catgut embedding can improve the symptoms of rheumatoid arthritis, and has the effect of combined treatment with leflunomide. Its mechanism may be related to PD-1 / OX40 signal pathway.