摘要
目的:观察实验性关节炎大鼠应用中药痹肿消汤干预后,促进滑膜血管翳形成的细胞因子肿瘤坏死因子α与血管内皮生长因子表达的变化,并与甲氨喋呤做标准对照。方法:实验于2003-05/11在中西医结合研究所实验室进行。取健康SD大鼠95只,随机分为4组:①正常组(n=5):不进行任何处理,于第25天处死。②模型组(n=30):采用牛II型胶原与完全福氏佐剂注射免疫诱导大鼠实验性关节炎模型,不给药。③甲氨喋呤组(n=30):同前造模,造模后即灌服甲氨喋呤0.43mg/kg,每周1次。④痹肿消汤组(n=30):同前造模,造模后即灌服痹肿消汤(白花蛇舌草、肿节风、薏米、络石藤、丹参、骨碎补等组成,2.07g/mL)10mL/kg,2次/d。后3组于造模后21d重复免疫1次,在造模后第25,30,35,40,45天分别处死6只大鼠,各组大鼠不同时间点血浆肿瘤坏死因子α水平用放射免疫法测定,滑膜组织血管内皮生长因子表达水平以免疫组化法检测,两种因子之间的关系用直线相关分析。结果:纳入的95只大鼠全部进入结果分析。①血浆肿瘤坏死因子α水平:造模后第25天模型组、痹肿消汤组与甲氨喋呤组均明显高于正常组犤(51.90±6.02),(32.68±8.76),(41.10±6.87),(10.37±1.29)μg/L,P<0.01犦;模型组高于痹肿消汤组及甲氨喋呤组(P<0.01或0.05),甲氨喋呤组高于痹肿消汤组(P<0.05)。随着时间的延长,模型组肿瘤坏死因子α水平逐渐升高,而痹肿消汤组及甲氨喋呤组则逐渐下降,且痹肿消汤组降低较甲氨喋呤组明显(P<0.05)。②滑膜血管内皮生长因子阳性细胞表达:正常组未见,其他3组均可见。在各时间点,模型组均明显高于痹肿消汤组和甲氨喋呤组(P<0.01),随着时间延长,症状加重,其表达逐渐升高;而痹肿消汤组和甲氨喋呤组则逐渐降低,且痹肿消汤组降低较甲氨喋呤组明显(P<0.05)。③肿瘤坏死因子α和血管内皮生长因子的相关性:两者呈正相关(r=0.893,2.478,P<0.01,0.05)。结论:痹肿消汤可降低关节炎大鼠血浆中肿瘤坏死因子α和关节滑膜组织中血管内皮生长因子的表达,从而阻抑滑膜血管翳形成;且痹肿消汤阻断肿瘤坏死因子α和血管内皮生长因子的作用优于甲氨喋呤。
AIM： To observe the changes of the expressions of cytokines of tumor necrosis factor-or in plasma and vascular endothelial growth factor, which can accelerate the formation of synovial pannus; in experimental rats with arthritis after intervention of the Chinese herb of bizhongxiao decoction, and compare with methotrexat. METHODS： The experiment was carried out in the laboratory of Institute of Integrated Chinese and Westem Medicine from May to November 2003. Ninety-five healthy SD rats were randomly divided into 4 groups： ① normal group （n=5）： the rats did not receive any treatment, and were killed/on the 25^th day; ② model group （n=30）： the rats were immunized and induced to models of arthritis by subcutaneous injection of collagen Ⅱ and complete Feud＇s adjutant, and they were not administrated. ③ methotrexat group （n=30）： modal establishment was the same as that mentioned above, and then the rats were perfused with methotrexat （0.43 mg/kg）, once a week; ④bizhongxiao decoction group （n=30）： model establishment was the same as that mentioned above, and then the rats were perfused with 10 mL/kg bizhongxiao decoction （composed of spreading hedyotis herb, glabrous sarcandra herb, coix seed, confederate-jasmine stem, danshen root and fortune＇s drynaria rhizome, 2.07 g/mL）, twice a day. The rats in the latter 3 groups were immunized again on the 21^st day after model establishment, and then 6 rats were killed respectively in each group on the 25^th 30^th, 35^th, 40^th and 45^th days after model establishment, the level of tumor necrosis factor-α in plasma at different time point was determined with radioimmunoassay, the expressive level of vascular endothelial growth factor in synovial tissue was detected with immunohistoassay, and the association between the two kinds of cytokines was analyzed with the linear correlation analysis. RESULTS： All the 95 rats were involved in the analysis of results. ① The level of tumor necrosis factor-α in plasma： On the 25^th day after model establishment, it was obviously higher in the model group, bizhongxiao decoction＇group and methotrexat group than in the normal group [（51.90±6.02）, （32.68±8.76）, （41.10±6.87）, （10.37±1.29） μg/L, P 〈 0.01], higher in the model group than in the bizhongxiao decoction group and methotrexat group （P 〈 0.01 or 0.05）, also higher in the methotrexat group than in the bizhongxiao decoction group （P 〈 0.05）. With the prolongation of time, the level of tumor necrosis factor-α was gradually increased in the model group, but gradually decreased in the bizhongxiao decoction group and methotrexat group, and the decrease was more obvious in the bizhongxiao decoction group than in the methotrexat group （P 〈 0.05）. ② The expression of positive cells of vascular endothelial growth factor in synovial tissue： The expression was not observed in the normal group, but observed in the other 3 groups. It was obviously higher in the model groupthan in the bizhongxiao decoction group and methotrexat group at each time point （P 〈 0.01）; As time prolonged, the symptoms were aggravated, and the expression was gradually increased in the model group, but gradually decreased in the bizhongxiao decoction group and methotrexat group, and the decrease was more obvious in the bizhongxiao decoction group than in the methotrexat group （P 〈 0.05）. ③ The association between tumor necrosis factor-or and vascular endothelial growth factor：. There was positive correlation between them （r=0.893, 2.478, P 〈 0.01, 0.05）. CONCLUSION： Bizhongxiao decoction can decrease the expressions of tumor necrosis faetor-a in plasma and vascular endothelial growth factor in the synovial tissue of joint in rats with arthritis, and then inhibit the formation of synovial pannus. Bizhongxiao decoction hasa better effect in inhibiting tumor necrosis factor-α and vascular endothelial growth factor than methotrexat.
出处
《中国临床康复》
CSCD
北大核心
2005年第27期118-120,共3页
Chinese Journal of Clinical Rehabilitation
基金
国家自然科学基金资助课题(30371833)
湖南省科技基金(02SSY4028)
湖南省中医管理局(20053)~~
