祛痰化瘀利湿方对大鼠膝骨关节炎滑膜组织和血清IL-1β,MMP-1及COMP,CTX-Ⅱ的影响  

The Effect of Qutanhuayulishi Decoction on Synocial IL-1β,MMP-1 and Serum IL-1β,MMP-1,COMP and CTX-Ⅱin Rats with Knee Osteoarthritis

作　　者：姚丽[1] 赵婧[1] 周强[1] 阚卫兵[1] 肖志锋[1] 王德刚[1] 张玉婷[1] 姜玉祥[1] 谢殿洪[1] 于为国[1] 

机构地区：[1]上海中医药大学附属普陀医院伤骨科,上海200062

出　　处：《中国中医骨伤科杂志》2015年第11期5-8,14,共5页Chinese Journal of Traditional Medical Traumatology & Orthopedics

基　　金：国家中医药管理局中医药重点学科建设项目-中医全科基金(2012;32号);上海市卫生局课题资助(20114059);上海市教委创新课题资助(14YZ058);上海中医药大学预算内科研项目(2014YSN64)

摘　　要：目的:观察祛痰化瘀利湿方对大鼠膝骨关节炎(OA)模型关节滑膜组织和血清IL-1β,MMP-1以及血清软骨降解标志物COMP,CTX-Ⅱ的影响。方法:采用Hulth方法建立膝骨关节炎大鼠模型,造模成功后,将造模组随机分为模型组和祛痰化瘀利湿方组。正常组和模型组大鼠予以生理盐水灌胃,祛痰化瘀利湿方组予以祛痰化瘀利湿方中药灌胃,连续灌胃4周,腹主动脉取血方法抽取大鼠血液,离心后获取大鼠血清;外科手术方法打开大鼠膝关节,摘取滑膜组织。免疫印迹法(Western blot)观察大鼠膝关节滑膜组织中IL-1β,MMP-1的表达,采用酶联免疫吸附法(ELISA)检测大鼠血清IL-1β,MMP-1,COMP,CTX-Ⅱ的表达。结果:成功建立大鼠膝骨关节炎模型,免疫印迹法结果显示模型组大鼠滑膜组织中IL-1β,MMP-1的蛋白表达显著高于正常组,两组比较差异有统计学意义(P<0.01);祛痰化瘀利湿方组对OA滑膜组织中IL-1β,MMP-1的蛋白表达有明显下调作用,与模型组相比较差异有统计学意义(P<0.01)。酶联免疫吸附法结果显示模型组大鼠血清中IL-1β,MMP-1,COMP,CTX-Ⅱ的蛋白表达显著高于正常组,两组比较差异有统计学意义(P<0.01);祛痰化瘀利湿方组对血清中IL-1β,MMP-1,COMP,CTX-Ⅱ的蛋白表达有明显下调作用,与模型组相比较差异有统计学意义(P<0.01)。结论:1)大鼠膝骨关节炎模型膝关节滑膜组织和血清中IL-1β,MMP-1的表达升高,提示该变化与滑膜炎症有关,MMP-1表达的升高参与诱导了软骨降解;而血清中COMP,CTX-Ⅱ的升高提示软骨降解,符合骨关节炎的特征。2)祛痰化瘀利湿方对IL-1β,MMP-1,COMP,CTX-Ⅱ的表达均有下调作用,提示祛痰化瘀利湿方能够抑制滑膜炎症,抑制滑膜炎症诱导的软骨降解,促进软骨的修复,这是祛痰化瘀利湿方治疗膝骨关节炎的机制之一。Objective.. To detect the effect of Qutanhuayulishi decoction on synoeial IL-1β, MMP-1 and serum IL-1β, MMP-1, COMP and CTX-Ⅱ in rats with knee osteoarthritis. Methods： The rats＇ model of knee osteoarthritis were established by Hulth methods, and they were randomly divided into model group and Qutanhuayulishi decoction group. The rats were intragastrie administrated with normal saline in the model group and normal group; and were intragastric administrated with Qutanhuayulishi decoction in the Qutanhuayulishi decoction group, for 4 weeks. The blood was got from abdominal aortic vessel and centrifuged to obtain serum sample. The synovial tissue of knee was obtained though surgery. The expression of the synocial IL-1β and MMP- 1 were detected by Western blot method, and the serum IL-1β, MMP-1, COMP and CTX-Ⅱ were detected by enzyme-linked im-unosorbent assay （ELISA） method. Results： The rats＇ model of knee osteoarthritis was established successfully. The results of Western blot showed that the expression of the synocial IL-1β and MMP-1 in the model group was significantly higher than normal group （P〈0.01）. The expression of the synocial IL-1β and MMP-1 in the Qutanhuayulishi decoction group was significantly decreased than the model group （P〈0.01）. The results of ELISA show that the expression of serum IL-1β, MMP-1, COMP and CTX-Ⅱ were higher than the normal group （P〈0.01）. The expression of the serum IL-1β, MMP-1, COMP and CTX-Ⅱ in the Qutanhuayulishi decoction group was significantly decreased than the model group （P〈0.01）. Conclusion： 1 ）The higher expression of IL-1β and MMP-1 in synovial tissue and serum of rats with knee osteoarthritis may suggest that this change may be related to inflammation of the synovial. The increasing expression of MMP-1 may be involved in the degration of cartilage, while the increasing expression of COMP, CTX-Ⅱ in serum imply cartilage degradation, which meets the characteristics of osteoarthritis. 2）Qutanhuayulishi decoct

关 键 词：膝骨关节炎 祛痰化瘀利湿方 白介素-1β(IL-1β) 基质金属蛋白酶-1(MMP-1) 软骨寡聚基 质蛋白(COMP) Ⅱ型胶原C-端肽(CTX-Ⅱ) 

分 类 号：R-33[医药卫生]