小花清风藤醇提液对RA大鼠相关蛋白的影响

Effect of Sabia parviflora ethanol extract on related proteins in rheumatoid arthritis rats

目的:探讨小花清风藤醇提液对类风湿关节炎(Rheumatoid arthritis,RA)的影响及对TNF-ɑ、HIF-1α、MMP-3及MMP-9表达的影响。方法:选择60只SPF级雄性Wistar大鼠,随机分为6组。除正常组外,余5组大鼠建立RA模型20d。而后灌胃给药干预20d,其中分别给予小花清风藤醇提液120mg/kg、90mg/kg、60mg/kg、阳性对照地塞米松7. 5mg/kg,模型组予等体积生理盐水。40d后处死大鼠,取血清、膝关节滑膜,分析各组大鼠足爪图片、X射线影像及足跖肿胀度;膝关节滑膜HE染色分析组织形态改变;免疫组化检测滑膜中MMP-9、HIF-1α、MMP-3及TNF-ɑ的表达变化; ELISA法检测血清中TNF-ɑ、HIF-1α、MMP-3及MMP-9的含量。结果:模型组较正常组大鼠足爪见明显肿胀、畸形及溶骨现象,给药后各组症状均有减轻,正常组未见异常; HE染色显示,正常组大鼠滑膜组织形态正常,模型组大鼠滑膜组织中有大量炎细胞浸润、血管翳增生,给药后各组炎细胞浸润和血管翳增生均有减少,尤其是90mg/kg、120mg/kg组;模型组血清中TNF-ɑ、HIF-1α、MMP-3及MMP-9的表达均显著高于正常组,小花清风藤组和阳性组较模型组均有明显减少,尤其是90mg/kg组;免疫组化结果显示,模型组大鼠滑膜组织中TNF-α、HIF-1α、MMP-9及MMP-3的阳性表达均显著升高,用药后各组较之模型组均显著下降,尤其是90mg/kg组。结论:小花清风藤醇提液对RA具有治疗作用,其机制可能与下调HIF-1α、MMP-3、MMP-9、及TNF-ɑ表达,抑制炎症发展与血管新生相关。

Objective: To explore the effect of Sabia parviflora ethanol extract on the expressions of tumor necrosis factor-α( TNF-α),hypoxia-inducible factor-1ɑ( HIF-1α),matrix metalloproteinase-3( MMP-3) and matrix metalloproteinase-9( MMP-9) in rheumatoid arthritis( RA) rats and investigate its mechanism. Methods: Sixty SPF male Wistar rats were randomly divided into the control group,the model group,120mg/kg high dose group,90mg/kg medium dose group,60mg/kg low dose group and 7. 5 mg/kg dexamethasone positive control group. RA model was established in the first 20 days. After that,rats in all doses of Sabia parviflora ethanol extract groups and dexamethasone group were intragastrically administered for constantly 20 days,other rats were administered with same volume of saline( 2 ml),once daily. After the last administration,all animals were executed to analyze the appearance of paws by X-ray and macroscopic observation. The contents of TNF-α,HIF-1α,MMP-3 and MMP-9 in serum and the synovial tissues were measured by ELISA and immunohistochemistry,respectively,and the tissue morphology of the synovial tissues were observed after HE staining under microscope. Results: In the model group,soft tissue swelling,joint deformity and osteolysis of rat paws were more obvious than those in the control group,all symptoms were alleviated in Sabia parviflora groups and positive control group. The synovial tissues of rats in the control group showed a normal morphology in HE straining,while inflammatory cells and pannus hyperplasia were observed in the model group. Both inflammatory cell infiltration and pannus hyperplasia were alleviated after drug administration in high dose group. The serum levels of the TNF-α,HIF-1α,MMP-3 and MMP-9 were significantly increased in the model group,and was decreased in Sabia parviflora groups and positive control group,there was the significant difference in 90mg/kg dose group( P < 0. 05). In immunohistochemical staining,expressions of TNF-α、HIF-1α、MMP-9 and MMP-3 in the model group were significantly enhanced than those in the control group,and was significantly decreased after drug administration. Conclusions: Sabia parviflora has therapeutic effect on RA,the mechanism may be related to down-regulating expressions of TNF-α,HIF-1α,MMP-3 and MMP-9,inhibition of inflammation and angiogenesis.