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目的观察祛痹方对Ⅱ型胶原诱导的免疫性关节炎模型大鼠氧自由基代谢物超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)及低氧诱导因子(hypoxia-inducible factor,HIF)-1α的影响,探讨祛痹方对类风湿关节炎(rheumatoid arthritis,RA)的治疗作用及其机制。方法选用SPF级雄性SD大鼠72只,随机选取1 2只作为空白对照组,其余60只采用尾根部皮下注射Ⅱ型胶原与不完全弗氏佐剂的方法 诱导制备关节炎模型,免疫15天后,将造模大鼠随机分为胶原诱导性关节炎(collagen-induced arthritis,CIA)模型组、雷公藤多苷治疗组、祛痹方高剂量及低剂量组,每组15只,予以灌胃给药,祛痹方高剂量组:6.66 g/(kg·d);祛痹方低剂量组:3.33 g/(kg·d);雷公藤多苷组:9.68 mg/(kg·d);空白对照组和CIA模型组大鼠灌服等体积纯净水。每日1次,连续4周。饲养期间测量大鼠踝关节肿胀度。治疗4周后处死动物,采用比色法测定血浆中SOD、MDA和GSH-P×活性;采用免疫组化法检测大鼠膝关节HIF-1α的表达。结果 (1)与CIA模型组比较,祛痹方高剂量组与雷公藤多苷组大鼠关节肿胀度均显著减轻(P<0.01,P<0.05),且祛痹方高剂量组关节肿胀度明显轻于雷公藤多苷组(P<0.05);(2)与CIA模型组比较,雷公藤多苷组及祛痹方高、低剂量组大鼠血浆GSH-P×活性明显升高,MDA活性降低,雷公藤多苷和祛痹方高剂量组大鼠血浆SOD活性明显升高(均P<0.05);(3)与CIA模型组比较,祛痹方高剂量组及雷公藤多苷组大鼠膝关节HIF-1α表达水平明显降低(P<0.05),祛痹方低剂量组有下降趋势,但差异无统计学意义(P>0.05)。结论祛痹方可显著减轻CIA模型大鼠踝关节肿胀程度,其疗效优于雷公藤多苷。其作用机制可能是通过下调关节HIF-1α表达及调节抗氧化水平,从而达到治疗CIA的作用。
Objective To observe the effect of Qu-Bi-Fang on superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidation in type 2 collagen induced immune arthritis rats. (GSH-Px) and hypoxia-inducible factor (HIF) -1α in patients with rheumatoid arthritis (RA) and to explore the therapeutic effect and mechanism of Qubi Recipe on rheumatoid arthritis (RA). Methods Seventy two Sprague-Dawley male Sprague-Dawley rats were randomly divided into two groups. One hundred and two male Sprague-Dawley rats were selected randomly as the blank control group. The remaining 60 rats were induced with arthritis by subcutaneous injection of type Ⅱ collagen and incomplete Freund’s adjuvant. The model rats were randomly divided into collagen-induced arthritis (CIA) model group, tripterygium glycosides treatment group and Qubi Fang high-dose and low-dose group, 15 rats in each group. The dosage of Qu Bi Fang: high dose group was 6.66 g / (kg · d); low dose group of Qu Bi Fang: 3.33 g / (kg · d); tripterygium glycosides group: 9.68 mg / (kg · d) The blank control group and CIA model group were fed with the same volume of pure water. 1 day, for 4 weeks. Rat ankle swelling was measured during feeding. After 4 weeks of treatment, the animals were sacrificed, and the activities of SOD, MDA and GSH-Px in the plasma were measured by colorimetric assay. The expression of HIF-1α in the knee joint was detected by immunohistochemistry. Results (1) Compared with CIA model group, the swelling degree of joint in high-dose Qubi Fang group and tripterygium glycosides group were significantly reduced (P <0.01, P <0.05), and Qubi Fang high dose group (P <0.05). (2) Compared with CIA model group, the activity of GSH-Px in plasma increased significantly in high and low doses of tripterygium glycosides group and Qubi Fang group, MDA (P <0.05). (3) Compared with CIA model group, high-dose Qubufang group and tripterygium glycosides group The expression level of HIF-1α in the knee joint of rats decreased significantly (P <0.05), while the low-dose Qubi Fang group showed a decreasing trend, but the difference was not statistically significant (P> 0.05). Conclusion Qubi Prescription can significantly reduce the degree of ankle swelling in CIA model rats, its efficacy is superior to Tripterygium glycosides. Its mechanism may be through the down-regulation of the expression of HIF-1α and regulate the level of anti-oxidation, so as to achieve the role of CIA.