Wu Yong-Ji, Zhang Hong-Tao, Zhang Xing-Hua, Ren Chao-Zhan, Jie Wang-Jun, Zhang Guang-Jun
College of Acupuncture-Moxibustion and Tuina, Gansu University of Chinese Medicine, Lanzhou 730000, China.
The 1st Department of Acupuncture and Moxibustion, Gansu Provincial Hospital of Traditional Chinese Medicine, Lanzhou 730050.
Zhen Ci Yan Jiu. 2025 Jul 25;50(7):808-814. doi: 10.13702/j.1000-0607.20240529.
To observe the effects of heat-reinforcing needling on inflammation and oxidative stress in rheumatoid arthritis (RA) model rats with cold syndrome, so as to analyze the mechanism of heat-reinforcing needling in improving RA.
Male SD rats were randomly divided into blank group, model group, heat-reinforcing needling group and resveratrol group according to random number table method, with 12 rats in each group. The rat model of RA with cold syndrome was established by subcutaneous injection of bovine type II collagen and complete Freund adjuvant combined with the stimulation of cold and wet environment. After successful modeling, rats of the resveratrol group was intraperitoneally injected with resveratrol (the agonist of Sirt1, 10 mg/kg), and rats of the heat-reinforcing needling group received heat-reinforcing needling at "Zusanli"(ST36)for 30 min every day. After 14 d of continuous intervention, the synovial morphology of knee joint was observed by HE staining. The activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), contents of reactive oxygen species (ROS) and malondialdehyde (MDA) in serum were detected by WST-1 method, colorimetric method probe method, and TBA method, respectively. The mRNA and protein expression levels of silencing information regulatory factor 1 (Sirt1) and (FoxO1) in the synovial tissue of knee joint were detected by Western blot and qPCR, respectively.
Compared with blank group, the fibroplasia, inflammatory cell infiltration, the relative expressions of Sirt1 mRNA and protein in synovial of knee joint, serum SOD and GSH-Px activity in model group were significantly decreased(<0.01), while the relative expressions of FoxO1 mRNA and protein, the contents of ROS and MDA were significantly increased(<0.01). After interventions, the decreased Sirt1 mRNA and protein, decreased SOD and GSH-Px activities, and the increased FoxO1 mRNA and protein, increased contents of ROS and MDA were reversed (<0.01, <0.05)in both of the heat-reinforcing needling and resveratrol groups. At the same time, the fibrous tissue proliferation and inflammatory cell infiltration around the synovial tissue of knee joint induced by RA were significantly reduced in both of the intervention groups.
Heat-reinforcing needling can inhibit oxidative stress injury and inflammation response in RA rats with cold syndrome, which may be related to the regulation of Sirt1/FoxO1 pathway.
观察温针治疗对寒证类风湿关节炎（RA）模型大鼠炎症及氧化应激的影响，以分析温针改善RA的作用机制。
将雄性SD大鼠按随机数字表法随机分为空白组、模型组、温针组和白藜芦醇组，每组12只。采用皮下注射牛Ⅱ型胶原和完全弗氏佐剂并结合寒湿环境刺激的方法建立寒证RA大鼠模型。造模成功后，白藜芦醇组大鼠腹腔注射白藜芦醇（Sirt1激动剂，10 mg/kg），温针组大鼠每日取“足三里”（ST36）行温针治疗30分钟。连续干预14天后，通过苏木精-伊红（HE）染色观察膝关节滑膜形态。分别采用WST-1法、比色法探针法和硫代巴比妥酸（TBA）法检测血清中超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）活性，活性氧（ROS）、丙二醛（MDA）含量。分别采用蛋白质免疫印迹法（Western blot）和实时荧光定量聚合酶链反应（qPCR）检测膝关节滑膜组织中沉默信息调节因子1（Sirt1）和叉头框蛋白O1（FoxO1）的mRNA和蛋白表达水平。
与空白组比较，模型组膝关节滑膜组织纤维组织增生、炎性细胞浸润，膝关节滑膜组织中Sirt1 mRNA和蛋白相对表达量、血清SOD和GSH-Px活性均显著降低（<0.01），而FoxO1 mRNA和蛋白相对表达量、ROS和MDA含量显著升高（<0.01）。干预后，温针组和白藜芦醇组大鼠膝关节滑膜组织中降低的Sirt1 mRNA和蛋白、降低的SOD和GSH-Px活性，以及升高的FoxO1 mRNA和蛋白、升高的ROS和MDA含量均得到逆转（<0.01，<0.05）。同时，两个干预组大鼠RA所致膝关节滑膜组织周围纤维组织增生和炎性细胞浸润均明显减轻。
温针可抑制寒证RA大鼠的氧化应激损伤和炎症反应，其机制可能与调控Sirt1/FoxO1信号通路有关。
