目的通过观察关节腔内注射臭氧对胶原诱导性关节炎(CIA)模型大鼠的干预疗效,血清核因子κB受体活化因子配体(RANKL)、护骨因子水平的影响,探讨臭氧治疗CIA大鼠的作用机制。方法将40只体质量及鼠龄相当的Wistar大鼠采用随机数字表法,随机分为健康对照组、CIA模型组(CIA组)、臭氧组、甲氨蝶呤组。除健康对照组外,采用注射弗氏完全佐剂+牛Ⅱ型胶原蛋白的方法建立大鼠CIA模型。造模成功后,臭氧组双踝关节内注射浓度为40μg/ml的臭氧各1ml,每周1次,共注射3周。甲氨蝶呤组腹腔注射甲氨蝶呤0.9mg/kg,每周1次,共3周。分别测量双足肿胀度、评估关节炎指数(AI)和影像学评分。治疗结束后1周,各组大鼠干预结束后内眦静脉取血,用流式多因子检测技术测定各组大鼠血清中护骨因子、RANKL的水平。应用重复测量方差分析或Wilcoxon秩和检验进行组间比较。结果①臭氧关节腔注射3周后大鼠双足肿胀度与AI较同期CIA组均有明显改善[足肿胀度:臭氧组(4.21±0.14)ml,CIA组:(9.12±0.17)ml,T=64.08,P=0.00;AI:臭氧组:(2.97±0.18)ml,CIA组:5.76±0.13,T=37.24,P=0.00],X线片显示关节破坏程度减轻。②CIA组血清中RANKL水平明显高于同期健康对照组[CIA组:1890.70(797.03,10571.94),健康对照组74.46(29.21,95.37),T=43,P=0.005],差异有统计学意义;臭氧组与同期CIA组相比RANKL水平明显降低[臭氧组28.09(14.11,207.30),CIA组:1890.70(797.03,10571.94),T=39,P〈0.0女],差异有统计学意义;臭氧组与甲氨蝶呤组相比RANKL水平相当,差异无统计学意义(T=52,P〉0.05])。③CIA组血清中RANKL/护骨因子明显高于同期健康对照组,差异有统计学意义[CIA组:250.68(42.33,2959.78),健康对照组4.32(3.16,5.30)?
Objective To evaluate the clinical efficacy of intra-articular injection of ozone in collagen- induced arthritis (CIA) rats and to assess its effects on serum receptor activator of nuclear factor κB ligand (RANKL) and osteoprotegerin (OPG) levels. Methods Forty weight age malched Wistar rats were randomly divided into normal control group (normal group), the CIA model group (CIA group), ozone (O3 group), andmethotrexate (MTX group). In addition to the normal control group, Freund's complete adjuvant and bovine type Ⅱ collagen were injected to establish the rat model of CIA. After the model was sucessfully developed, double ankle injection concentration ozone group of 40 μg/ml of O3 each 1 ml, 1 times a week, a total of injection for 3 weeks for the experimenal group. MTX group of 0.9 mg/kg was injected 1 times a week for 3 weeks for the MTX group. Degree of foot swelling was measured, and radiographic assessment of arthritis index (AI) score was performed. One week after treatment, angular vein blood was collected for the rats after the intervention, flow multi-factor detection technology was used to test each rat. T test or Wilcoxon rank test was used to compare the difference between groups. Results ① After 3 week administration with O3, dcgree of foot swelling, and AI of the O3 group was reduced significanly than the CIA group during the same period (foot swelling degree:O3 group: (4.21±0.14) ml, CIA group (9.12±0.17) ml, T=64.08, P=0.00; AI O3 group: [(2.97± 0.18) ml, CIA group: 5.76±0.13, T=37.24, P=0.00], and X-ray showed joint damage was alleviated. ② The serum level of RANKL in the CIA group was significantly higher than of the normal group [CIA model group 1 890.70(797.O3, 10 571.94)], normal group [74.46(29.21, 95.37), T=43, P=0.005] during the same period; The serum level of RANKL in the O3 group was significantly lower than the CIA group [O3 group 28.09(14.11, 207.30), CIA group 1 890.70 (797.O3, 10 571.94), T=39, P〈