胶原诱导型关节炎模型大鼠血浆血管内皮生长因子与肿瘤坏死因子α表达及反应停的干预效应

Thalidomide interferes the expressions of vascular endothelial growth factor and tumor necrosis factor-alpha in rats with collagen-induced arthritis

目的:观察胶原诱导型关节炎大鼠病程及致病过程中两种重要炎性因子血管内皮生长因子与肿瘤坏死因子α在反应停干预后的影响。方法:实验于2006-01/07在河北医科大学第二医院完成。成年雄性Wistar大鼠112只,体质量140～160g,随机数字表法分为为正常对照组、单纯造模组、反应停治疗组和甲氨蝶呤治疗组,每组28只。除正常对照组外,其余3组多点皮内注射Ⅱ型胶原与完全弗氏佐剂的乳化剂,诱导出关节炎模型。正常对照组作为阴性对照组每只大鼠背部及尾根部多点皮内注射等量的生理盐水。从免疫后第10天开始,正常对照组和单纯造模组大鼠给予蒸馏水0.5mL/(只·d)灌胃;反应停治疗组给予反应停200mg/(kg·d)灌胃;甲氨蝶呤治疗组给予甲氨蝶呤2.7mg/(kg·周)灌胃。测定造模后7,14,21,28,35,42,60d不同时间点各组大鼠的足爪厚度、血浆肿瘤坏死因子α及血管内皮生长因子的水平,进行统计学分析。结果:纳入大鼠112只,均进入结果分析。①各组大鼠足爪肿胀程度:实验前各组大鼠之间足爪厚度无显著差异[(5.47±0.14),(5.33±0.10),(5.92±0.24),(5.35±0.23)mm,P>0.05]。与正常对照组比较,单纯造模组、反应停治疗组及甲氨蝶呤治疗组大鼠足爪厚度均于造模后7d开始增高,21d左右达到肿胀高峰,差异有显著性意义[7d:(5.42±0.24),(5.81±0.15),(6.59±0.31),(5.54±0.33)mm;21d:(5.73±0.23),(10.78±0.53),(9.82±0.34),(10.37±0.57)mm,P<0.01]②各组大鼠血浆肿瘤坏死因子α及血管内皮生长因子水平及两者关系:实验前各组血浆肿瘤坏死因子α及血管内皮生长因子水平均无显著差异[(2.351±0.208),(2.331±0.213),(2.280±0.171),(2.283±0.125)nmol/L;(1.807±0.094),(1.811±0.099),(1.819±0.101),(1.813±0.090)nmol/L;P>0.05],与正常对照组相比,单纯造模组、反应停治疗组及甲氨蝶呤治疗组大鼠两者水平于造模后7d开始增高,差异具有显著性意义[(2.245±0.234),(2.450±0.026),(2.419±0.069),(2.467±0.032)nmol/L;(1.803±0.099),(2.024±0.049),(2.017±0.101),(2.055±0.056)nmol/L;P<0.05]。两者水平呈直线相关。结论:反应停可以降低关节炎模型大鼠血浆肿瘤坏死因子α及血管内皮生长因子浓度,可以有效减轻其足爪肿胀程度,疗效与甲氨蝶呤相同。

AIM： To observe the effects of thalidomide on vascular endothelial growth factor （VEGF） and tumor necrosis factor α （TNF-α） as two important inflammatory factors in the course of collagen-induced arthritis of rats. METHODS： The experiment was performed in the Second Hospital of Hebei Medical University from January to July 2006. 112 adult male Wistar rats of 140-160 g were randomly divided into normal group, model group, thalidomide group and methotrexate （MTX） group with 28 rats in each group. All rats except the normal group were intradermally injected with the emulsion of collagen 11 and complete Freund＇s adjuvant to induce arthritis models. The normal group as negative control was intradermally injected matching normal sodium in the back and tail. From the tenth day, rats of the normal group and model group were given an intragastric administration with 0.5 mL daily distilled water; rats of the thalidomide group with 200 mg/kg daily thalidomide, and rats of the MTX group with 2.7 mg/kg per week MTX. The thickness of claws, and the concentration of TNF-α and VEGF were measured on 7, 14, 21, 28, 35, 42, and 60 days after modeling for statistical analysis. RESULTS： A total of 112 rats were involved in toe result analysis. ①Claw swelling of rats： There were no obvious differences in the thickness of claw before experiment [（5.47±0.14）, （5.33±0.10）, （5.92±0.24）, （5.35±0.23） mm, P 〉 0.05]. Compared with the normal control group, the thickness of claw of the model group, thalidomide group and MTX group began increase from the 7^th day after modeling, and reached peak at day 21, which had significant differences [day 7： （5.42±0.24）, （5.81±0.15）, （6.59±0.31）, （5.54±0.33） mm; day 21： （5.73±0.23）, （10.78±0.53）, （9.82±0.34）, （10.37±0.57） mm, P 〈 0.01]. ②Association between TNF-α and VEGF： There were no significant differences in the concentration of TNF-α and VEGF among each group [（2.351±0.208）, （2.331±0.213）, （2.280±0.171）, （2.283±0.125） nmol/L, （1.807±0.094）, （1.811± 0.099）, （1.819±0.101）, （1.813±0.090） nmol/L;P 〉 0.05]. Compared with the normal control group, the TNF-α and VEGF levels of the model group, thalidomide group and MTX group began increase from the 7th day after modeling, and the differences were significant [（2.245±0.234）, （2.450±0.026）, （2.419±0.069）, （2.467±0.032） nmol/L, （1.803±0.099）, （2.024± 0.049）, （2.017±0.101）, （2.055±0.056） nmol/L;P 〈 0.05]. The levels of TNF-α and VEGF were related linearly. CONCLUSION： Thalidomide could reduce the concentration of TNF-α and VEGF in blood plasma and lessen effectively the early swelling of rats with collagen-induced arthritis, which displays the same effect as MTX.