关节内注射不同浓度臭氧对类风湿关节炎大鼠TNF-α、TNFRⅠ和TNFRⅡ的影响

Effect of intra-articular ozone injection on serum and synovial TNF-α, TNFR I, and TNFR II contents in rats with rheumatoid arthritis

目的观察不同浓度臭氧(O3)对类风湿关节炎(RA)大鼠体内TNF-α、TNFRⅠ和TNFRⅡ的影响,阐述O3治疗类风湿关节炎的机理。方法健康雄性Wistar大鼠48只,采用随机数字表法分为CON组、RA组、O2组、O3-10组、O3-20组、O3-30组、O3-40组、O3-50组,每组6只。除空白对照组外,采用注射弗氏完全佐剂乳化的牛Ⅱ型胶原蛋白的方法建立大鼠RA模型。造模后21d,根据分组关节内注射纯氧和浓度分别为10、20、30、40、50μg/ml的O3各1ml,每周1次,共注射3周。治疗前后测量双后肢足爪厚度。治疗结束后1周,检测大鼠血清和滑膜TNF-α、TNFRⅠ和TNFRⅡ的含量。结果治疗结束后,O3-40组大鼠足爪厚度减小,与RA组相比差异有统计学意义(P<0.01)。RA组、O2组、及O3治疗各组血清中TNF-α、TNFRⅠ、TNFRⅡ含量差异无统计学意义(P>0.05)。与RA组相比,O3-40组、O3-50组大鼠滑膜TNF-α、TNFⅡ含量降低,差异有统计学意义(P<0.01)。与RA组相比,O3-40组大鼠滑膜TNFRⅠ含量升高,差异有统计学意义(P<0.05)。结论关节内注射O3可减轻RA大鼠的关节肿胀,浓度为40μg/ml的O3最为明显,作用机制和O3可以降低滑膜内TNF-α、TNFRⅡ的活性,上调TNFRⅠ的表达有关。

Objective To observe the effects of intra-articular ozone injection at different concentrations on the contents of tumor necrosis factor-α （TNF-α）, TNF receptor I （TNFR I）, and TNFR II in the serum and synovium of rats with rheumatoid arthritis （RA） and explore the therapeutic mechanism of ozone in RA treatment. Methods Forty-eight Wistar rats were randomized into 8 groups, including 5 ozone groups receiving intra-articular injection of 10, 20, 30, 40 or 50 μg/ml ozone, a blank control group, an oxygen group and a RA model group. All the rats, except for those in the blank control group, were subjected to hypodermic injection of bovine collagen II and complete Freund’s adjuvant to induce RA. Ozone treatment was administered once weekly for 3 weeks starting at 21 days after the modeling. The swelling and thickness of the hind paws were observed, and the serum and synovial contents of TNF-α, TNFR I, and TNFR II were detected. Results At the end of treatment, the paw thickness was reduced significantly in rats with 40 μg/ml ozone injection compared with that in the model RA group （P〈0.01）. The serum contents of TNF-α, TNFR I and TNFR II showed no significant difference between the RA model group, oxygen group and the ozone groups, but their synovial contents showed significant reductions in rats with 40 and 50 μg/ml ozone injection （P〈0.01）; the synovial TNFR I was significantly higher in 40 μg/ml ozone group than in the model group （P〈0.05）. Conclusion Intra-articular injection of 40 μg/ml ozone can attenuate synovitis in rats with RA, the mechanism of which may involve the inhibition of TNF-α and TNFR II activity and enhancement of TNFR I activity in the synovium