雷公藤内酯醇对胶原诱导关节炎大鼠外周血清和关节腔中细胞因子水平的干预(英文)

Interventional effects of triptolide on the levels of cytokines in peripheral serum and articular cavity of rats with collagen induced arthritis

背景:雷公藤具有抗炎、免疫抑制等作用,目前已被用于治疗类风湿关节炎等多种自身免疫性疾病。其化学成分复杂,其中雷公藤内酯醇被公认为是雷公藤中重要的有效成分之一。目的:建立Ⅱ型胶原诱导的关节炎大鼠模型,观察雷公藤内酯醇对其外周血清和关节腔中白细胞介素6、白细胞介素10以及肿瘤坏死因子α含量的影响。设计:随机对照动物实验。单位:华中科技大学同济医学院附属同济医院中西医结合科。材料:实验于2004-11/2005-07在武汉同济医院完成。选取清洁级健康雄性Wistar大白鼠50只(购于华中科技大学同济医学院实验动物中心,动物质量合格证号:scxk(鄂)2004-2007)。雷公藤内酯醇(购于福建省医学科学研究所,临时生产无批号),纯度为98.5%以上。方法:①50只大鼠随机数字表法选10只作为正常对照组,其余建立关节炎模型。沿造模大鼠背部及尾根部分5点皮内注射Ⅱ型胶原乳剂,0.05mL/点,15d后分两点皮内激发注射。正常对照组予以生理盐水同法注射。初次免疫30d后参照关节炎指数评分标准对造模效果进行评估,共20只大鼠评分在6分以上成功造模而用于实验,随机数字表法分为关节炎模型组、雷公藤内酯醇组,10只/组。②将雷公藤内酯醇用体积分数为0.05的丙二醇配制成注射液,其中含雷公藤内酯醇100mg/L,雷公藤内酯醇组按0.04mL/100g后腿肌注给药,每3d用药1次,共30d。正常对照组给予等量生理盐水,关节炎模型组给予等量的体积分数为0.05的丙二醇液。③给药30d后取材,采用酶联免疫吸附法检测各组大鼠血清和膝关节腔液中白细胞介素6,10以及肿瘤坏死因子α的含量。主要观察指标:雷公藤内酯醇对外周血和关节腔液中肿瘤坏死因子α、白细胞介素6,10含量的影响。结果:实验选取清洁级雄性Wistar大鼠50只,10只作为正常对照,剩余40只大鼠共20只造模成功而进入结果分析。①关节炎模型组外周血和关节腔内的肿瘤坏死因子α含量最高,经雷公藤内酯醇治疗后均明显下降[(35.09±8.82),(15.35±3.56)ng/L;(44.17±8.94),(22.54±4.76)ng/L;P均<0.01],且与正常对照组基本相似(P>0.05)。②关节炎模型组外周血和关节腔内的白细胞介素6含量最高,经雷公藤内酯醇治疗后均明显下降[(76.58±6.81),(42.45±5.72)ng/L;(88.69±10.56),(48.67±5.97)ng/L;P均<0.01],但未能达到正常对照组水平(P<0.05)。③关节炎模型组外周血和关节腔内的白细胞介素10含量最低,经雷公藤内酯醇治疗后均明显升高[(17.53±2.07),(21.23±2.91)ng/L;(10.59±2.96),(14.74±1.85)ng/L;P均<0.01],且与正常对照组基本相似(P>0.05)。结论:雷公藤内酯醇可能通过对细胞因子含量的调节治疗关节炎。

BACKGROUND： Common threewingnut root has the functions of anti-inflammation and immune inhibition, etc., and it has been used at present to treat various autoimmune diseases including rheumatoid arthritis.Common threewingnut root has complex components, and triptolide is acknowledged as one of the important effective components of common threewingnut root. OBJECTIVE： To establish rat models of type 11 collagen induced arthritis, and observe the effects of triptolide on the contents of interleukin-6, interleukin-10 and tumor necrosis factor alpha （TNF-α） in peripheral serum and synovial fluid. DESIGN： A randomized control animal experiment. SETTING： Department of Integrated Traditional and Western Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology. MATERIALS： The experiments were carried out in the Tongji Hospital from November 2004 to July 2005. Fifty healthy male Wistar rats of clean degree were purchased from the experimental animal center of Tongji Medical College, Huazhong University of Science and Technology [qualification number of animal [scxk（E）2004-2007]. Triptolide （no batch number because of temporary production） was bought from Fujian Institute of Medical Sciences, and the purity was above 98.5%. METHODS： ①Ten of the 50 rats were randomly selected as the normal controls, and the others were made into models. Type Ⅱ collagen emulsion was injected intradermally at five points along the back and tail of the rats, 0.05 mL for each point, and injected intradermally at two points after 15 days. The rats in the normal control group were treated with saline in the same way. The effects of the model establishment were evaluated according to the scoring standards of arthritis index at 30 days after the first immunity, and the rats scored 6 points or above were taken as successful models and enrolled in the experiments. Twenty successful rat models were randomly divided into arthritis model group （n=10） and triptolide treated group （n=10）. ② Triptolide （100 mg/L） was dispensed into parenteral solution with propylene glycol （0.05 in volume fraction）, and then intramuscularly injected into hindlimb of rats in the triptolide treated group （0.04 mL/100 g）, once every three days for 30 days. The rats in the normal control group were given isovolume saline, and those in the arthritis model group were treated with isovolume propylene glycol （0.05 in volume fraction）. ③The materials were removed at 30 days after administration. The contents of interleukin-6, interleukin-10 and TNF-α in peripheral serum and synovial fluid were detected with enzyme-linked immunosorbant assay （ELISA）. MAIN OUTCOME MEASURES： The effects of triptolide on contents of TNF-α, interleukin-6 and interleukin-10 in peripheral serum and synovial fluid were observed. RESULTS： Fifty male Wistar rats of clean degree were selected, 10 were used as normal controls, and 20 of the other 40 rats were successfully made into models and enrolled in the analysis of results. ① The TNF-α contents in peripheral serum and synovial fluid were the highest in the arthritis model group, and obviously decreased after treatment of triptolide [（35.09±8.82）, （15.35±3.56） ng/L; （44.17±8.94）, （22.54±4.76） ng/L; P〈 0.01], which were similar to those in the normal control group （P 〉 0.05）. ② The contents of interleukin-6 in peripheral serum and synovial fluid were the highest in the arthritis model group, and were obviously decreased after treatment of triptolide [（76.58±6.81）, （42.45±5.72） ng/L; （88.69±10.56）, （48.67±5.97） ng/L; P 〈 0.01], but did not recover to the levels in the normal control group （P 〈 0.05）. ③The contents of interleukin-10 in peripheral serum and synovial fluid were the lowest in the arthritis model group, and obviously increased after treatment of triptolide [（17.53±2.07）, （21.23±2.91） ng/L; （10.59±2.96）, （14,74±1.85） ng/L; P 〈 0.01], which were similar to those in the normal control group （P 〉 0.05）. CONCLUSION： Triptolide can treat arthritis by modulating the contents of cytokines.