黄芪桂枝五物汤对阳虚寒凝型骨关节炎大鼠免疫相关细胞因子的影响  

Effect of Huangqi Guizhi Wuwu Tang on Immune Related Cytokines in Rats with Yang Deficiency and Cold Coagulation Type Osteoarthritis

作　　者：赵乐[1,2] 李艳彦[2] 王永辉[2] 周然[2] 

机构地区：[1]成都中医药大学,成都611137 [2]山西中医学院,太原030024

出　　处：《中国实验方剂学杂志》2017年第7期160-166,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：山西省科技厅攻关项目(2014011047-1);山西中医学院博士科研启动基金项目

摘　　要：目的:研究黄芪桂枝五物汤对阳虚寒凝型骨关节炎大鼠CD4^+/CD8`+及相关细胞因子的作用。方法:SD大鼠随机分为正常组、模型组、塞来昔布组、黄芪桂枝五物汤7 d组和黄芪桂枝五物汤14 d组,每组10只。除正常组外,各组大鼠均予冷固法复合外界环境刺激的方法复制阳虚寒凝型骨关节炎模型,共6周。造模结束前,各给药组分别灌胃给药,黄芪桂枝五物汤7 d组或14 d组分别给药7 d或14 d(30 g·kg-1),塞来昔布组给药7 d(20.82 mg·kg-1),正常组及模型组予生理盐水灌胃7 d,至造模结束。造模结束后,免疫组化法检测各组大鼠膝关节软骨诱导型一氧化氮合酶(i NOS),Ⅱ型胶原及转化生长因子-β1(TGF-β1)表达,取大鼠血液流式细胞法检测CD4^+/CD8^+,酶联免疫吸附测定(ELISA)法检测各组大鼠血液中白细胞介素-4(IL-4),C-反应蛋白(CRP)及胰岛素生长因子(IGF),低氧诱导因子1α(HIF-1α)含量。结果:与正常组比较,模型组CD4^+及CD4^+/CD8^+降低(P<0.05),各用药组与模型组比较,黄芪桂枝五物汤14 d组和塞来昔布组CD4+及CD4+/CD8+升高(P<0.05)。与正常组比较,模型组IL-4,CRP,HIF-1α及i NOS表达升高(P<0.05),Ⅱ型胶原及TGF-β表达降低(P<0.05);与模型组比较,各用药组Ⅱ型胶原及TGF-β表达升高(P<0.05),IL-4水平降低(P<0.05),黄芪桂枝五物汤14 d组CRP,i NOS及HIF-1α表达降低(P<0.05),塞来昔布组CRP及i NOS(P<0.05);各用药组之间,与塞来昔布组比较,黄芪桂枝五物汤7 d组和14 d组Ⅱ型胶原表达降低(P<0.05),黄芪桂枝五物汤7 d组i NOS表达升高(P<0.05)。IGF水平在各组无显著性差别。结论:黄芪桂枝五物可以调节机体免疫低下状态,并通过调节细胞因子IL-4及下游细胞因子i NOS,HIF-1α,TGF-β1等细胞因子水平的变化,对骨关节炎软骨细胞起正向修复作用。Objective： To study the effect of Huangqi Guizhi Wuwu Tang on CD4^＋/CD8^＋ and related cytokines in rats with Yang deficiency cold coagulation type osteoarthritis. Method： The 50 SD rats were randomly divided into normal group, model group, celecoxib group, Huangqi Guizhi Wuwu Tang 7 days group and Huangqi Guizhi Wuwu Tang 14 days group, n = 10 in each group. All the rats except those in the normal group received frozen and physical stimulation methods for 6 weeks to establish Yang deficiency cold coagulation type osteoarthritis models. Before successful modeling, the rats in all groupsreceived gastric administration of corresponding drugs or saline ： 7 days for Huangqi Guizhi Wuwu Tang 7 days group, 14 days for Huangqi Guizhi Wuwu Tang 14 days group （30 g·kg^-1） , and 7 days for Celecoxib group （20. 82 mg·kg^-1）. In addition, the rats in normal group and model group received normal saline for 7 days equally. After modeling, the expression levels of （iNOS） , collagen type Ⅱ and （TGF-β1） in knee joints were detected by immunohistochemical method. Blood was taken to test the levels of CD4 ^＋/CD8^＋ by using flow cytometry, and detect the levels of interleukin-4 （IL-4） , C-reactive protein （CRP） , insulinlike growth factors （IGF） and hypoxia inducible factor-1α（HIF-1α） by using ELISA method. Result： As compared with the normal group, the ratio of CD4^＋/CD8^＋ in model group was decreased （P 〈 0. 05）. As compared with the model group, the expression levels of CD4^＋ and the ratio of CD4^＋/CD8^＋ were increased in Huangqi Guizhi Wuwu Tang 14 days group and celecoxib group （P 〈 0.05）. As compared with the normal group, the expression levels of IL-4, CRP, iNOS and HIF-1α were increased in model group （P 〈 0.05）, while the expression levels of Collagen type 11 and TGF-β1 were decreased （P 〈 0.05）. As compared with the model group, the expression levels of Collagen type 11 and TGF-β1 were increased in various treatment groups （P 〈 0

关 键 词：黄芪桂枝五物汤 骨关节炎 免疫低下 细胞因子 

分 类 号：R285.5[医药卫生—中药学]