秦艽不同配伍对风寒湿痹模型大鼠MMP-3及TIMP-1的影响  

Effect of Different Combinations of Gentianae Macrophyllae Radix on MMP-3 and TIMP-1 in Rheumatoid Arthritis Rats with Wind-cold-dampness Arthralgia

作　　者：苏杉 王蓉[1] 赵敏[1] 吴晨 高慧琴[1] SU Shan;WANG Rong;ZHAO Min;WU Chen;GAO Hui-qin(Gansu University of Chinese Medicine,Lanzhou 730000,China)

机构地区：[1]甘肃中医药大学

出　　处：《中国实验方剂学杂志》2019年第3期8-14,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金地区项目(81360648)

摘　　要：目的:观察秦艽寒热不同配伍药对对风寒湿痹型类风湿关节炎(rheumatoid arthritis,RA)模型大鼠踝关节基质金属蛋白酶-3 (matrix metalloproteinase-3,MMP-3),基质金属蛋白酶组织抑制物-1 (matrix metalloproteinase tissue inhibitor-1,TIMP-1)的影响。方法:选取80只SD大鼠并随机分为正常组,Ⅱ型胶原组,风寒湿病证模型组,雷公藤多苷片组(6 mg·kg^-1),单味秦艽组(25 g·kg^-1),秦艽配伍威灵仙(秦威组,25 g·kg^-1),秦艽配伍桑寄生(秦桑组,25 g·kg^-1)、秦艽配伍防己(秦防组,25 g·kg^-1),共8组,每组10只。采用注射Ⅱ型胶原乳剂诱导并给予风寒湿刺激制备风寒湿RA大鼠模型。模型建立完成后正常组,Ⅱ型胶原组及风寒湿病证模型组大鼠给予生理盐水,各给药组给予相应药液灌胃。实验过程中,每3 d测量1次大鼠左后足跖厚度,计算足跖肿胀度;实验第38天对各组大鼠进行关节炎指数(AI)评分;酶联免疫吸附测定(ELISA)法检测大鼠血清类风湿因子(RF)含量;蛋白免疫印迹法(Western blot)检测踝关节MMP-3,TIMP-1的表达水平;实时荧光定量聚合酶链式反应(Real-time PCR)技术检测踝关节MMP-3,TIMP-1 mRNA的表达。结果:与正常组比较,Ⅱ型胶原组及风寒湿病证模型组足跖肿胀度,AI评分,血清RF含量,踝关节MMP-3蛋白表达水平,MMP-3 mRNA明显升高(P<0. 05,P<0. 01),踝关节TIMP-1蛋白表达水平,TIMP-1 mRNA显著降低(P<0. 01);与风寒湿病证模型组比较,各给药组足跖肿胀度,AI评分及血清RF含量均有不同程度降低,其中秦威组降低最显著(P<0. 01);Western blot及Real-time PCR结果显示,与风寒湿病证模型组比较,各给药组踝关节MMP-3蛋白表达水平及MMP-3 mRNA表达水平均显著降低(P<0. 01),以秦威组降低最明显,雷公藤多苷片组、秦威组及秦桑组大鼠踝关节TIMP-1蛋白表达水平及TIMP-1 mRNA表达水平均有所升高,以秦威组升高最显著(P<0. 01)。结论:对于风寒湿痹型类风湿关节炎,平温相�Objective: To observe the effects of different combinations of Gentianae Macrophyllae Radix( Qinjiao) on the ankle joint matrix metalloproteinase-3( MMP-3) and tissue inhibitor of matrix metalloproteinase-1( TIMP-1) of rheumatoid arthritis( RA) model rats with wind-cold-dampness arthralgia. Method: Eighty healthy SD rats were randomly divided into 8 groups,namely blank control group,collage Ⅱ model group,wind-colddampness syndrome model group, positive control group, single-taste Gentianae Macrophyllae Radix group,Gentianae Macrophyllae Radix-Clematidis Radix et Rhizoma group( GC group),Gentianae Macrophyllae RadixTaxilli Herba group( GT group),Gentianae Macrophyllae Radix-Stephanlae Tetrandrae Radix group( GS group),with 10 rats in each group. Rat model of wind-cold-dampness RA was induced through the injection with type Ⅱcollagen emulsion and wind-cold-dampness stimulation. After the establishment of the model,the blank control group,collage Ⅱ model group and wind-cold-dampness syndrome model group were given normal saline,and the corresponding liquid medicine was given to each administration group. In the experiment,the thickness of the left posterior metatarsal of rats was measured every 3 days,and the swelling degree of metatarsal was calculated. The arthritis index( AI) was evaluated on the 38 thday of the experiment. The serum rheamatoid factor( RF) content of rats was detected by enzyme linked immunosorbent assay( ELISA). The expressions of MMP-3 and TIMP-1 in ankle joint were detected by Western blot. The expressions of MMP-3 and TIMP-1 mRNA in ankle joint were detected by real-time fluorescence quantitative PCR( Real-time PCR). Result: Compared with the blank group,the swelling degree,AI score,serum RF content,MMP-3 protein expression and MMP-3 mRNA expression in ankle joints of coll age Ⅱ model group and model wind-cold-dampness syndrome group were significantly increased( P< 0. 05,P< 0. 01),while the TIMP-1 protein expression and TIMP-1 mRNA expression in ankle joints were significantly decrea

关 键 词：秦艽 风寒湿痹 类风湿关节炎 类风湿因子(RF) 基质金属蛋白酶-3(MMP-3) 基质金属蛋白酶组织抑制剂-1(TIMP-1) 

分 类 号：R20[医药卫生—中医学] R22