乌头汤联合甲氨蝶呤治疗大鼠类风湿关节炎的疗效及其与p38MAPK通路的相关性  

Effect of Wutou Decoction(乌头汤)Combined with Methotrexate on Rheumatoid Arthritis in Rats and Its Correlation with p38MAPK Pathway

作　　者：吴丹 李洁芳[1] 刘君 WU Dan;LI Jiefang;LIU Jun(The Fourth Hospital of Changsha,Changsha Hunan 410006,China)

机构地区：[1]长沙市第四医院,湖南长沙410006

出　　处：《中医药导报》2024年第2期26-31,共6页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：湖南省中医药科研计划项目(2019134)。

摘　　要：目的:探讨乌头汤(WTD)联合甲氨蝶呤治疗大鼠类风湿关节炎(RA)的效果及其与p38MAPK通路的相关性。方法:将75只雄性Wistar大鼠随机分为空白组、RA模型组、甲氨蝶呤组、WTD组、WTD+甲氨蝶呤组,每组15只。除空白组外,其余各组通过足跖面皮下注射弗氏佐剂及牛Ⅱ型胶原建立大鼠胶原性关节炎(CIA)模型。甲氨蝶呤组大鼠予甲氨蝶呤灌胃,WTD组予WTD灌胃,WTD+甲氨蝶呤组大鼠予WTD、甲氨蝶呤灌胃,空白组及RA模型组灌胃等量生理盐水。28 d后对大鼠踝关节结构和病理改变进行评分,采用聚合酶链反应(PCR)检测Ⅱ型胶原mRNA及p38MAPK mRNA表达水平,Western blotting法检测滑膜组织p38MAPK蛋白表达水平,酶联免疫吸附试验(ELIAS)检测血清TNF-α、IL-6水平。结果:RA模型组、甲氨蝶呤组、WTD组、WTD+甲氨蝶呤组大鼠Pelletier和Mankin评分均高于空白组(P<0.05);甲氨蝶呤组、WTD组、WTD+甲氨蝶呤组大鼠Pelletier和Mankin评分均低于RA模型组(P<0.05);WTD+甲氨蝶呤组大鼠Pelletier和Mankin评分低于WTD组、甲氨蝶呤组(P<0.05)。RA模型组大鼠软骨细胞Ⅱ型胶原mRNA相对表达量低于空白组(P<0.05),软骨细胞p38MAPK mRNA相对表达量、滑膜组织p38MAPK蛋白相对表达量及血清TNF-α、IL-6水平均高于空白组(P<0.05);甲氨蝶呤组、WTD组、WTD+甲氨蝶呤组大鼠软骨细胞Ⅱ型胶原mRNA相对表达量高于RA模型组(P<0.05),软骨细胞p38MAPK mRNA相对表达量、滑膜组织p38MAPK蛋白相对表达量及血清TNF-α、IL-6水平均低于RA模型组(P<0.05);WTD+甲氨蝶呤组大鼠软骨细胞Ⅱ型胶原mRNA相对表达量高于WTD组、甲氨蝶呤组(P<0.05),软骨细胞p38MAPK mRNA相对表达量、滑膜组织p38MAPK蛋白相对表达量及血清TNF-α、IL-6水平均低于WTD组、甲氨蝶呤组(P<0.05)。Pearson相关分析显示,各组大鼠血清TNF-α、IL-6水平与关节软骨中p38MAPK mRNA相对表达量呈正相关(P<0.05)。结论:WTD联合甲氨蝶呤治疗大�Objective:To investigate the effect of Wutou decoction(WTD)combined with methotrexate on rheumatoid arthritis(RA)in rats and its correlation with the p38MAPK pathway.Method:Totally 75 male Wistar rats were randomly divided into blank group,RA model group,methotrexate group,WTD group,and WTD+methotrexate group,with 15 rats in each group.Except for the blank group,the rats were established model of collagen induced arthritis(CIA)by subcutaneous injection of Freund's adjuvant and bovine typeⅡcollagen into the plantar surface in other groups.Rats were given intragastric administration of methotrexate in the methotrexate group,WTD in WTD group,WTD and methotrexatein in WTD+methotrexate group,and the same amount of physiological saline in blank group and RA group.The scores of ankle joint structure and pathological changes were evaluated in 28 days.Polymerase chain reaction(PCR)was used to detect the expression levels of typeⅡcollagen mRNA and p38MAPK mRNA in chondrocytes.Western blotting was used to detect the expression level of p38MAPK protein in synovial tissue,and enzyme linked immunosorbent assay(ELIAS)was used to detect serum TNF-αand IL-6 levels.Results:The RA model group,methotrexate group,WTD group,and WTD+methotrexate group showed higher Pelletier and Mankin scores than blank group(P<0.05);The methotrexate group,WTD group and WTD+methotrexate group showed lower Pelletier and Mankin scores than RA model group(P<0.05);The WTD+methotrexate group showed lower Pelletier and Mankin scores than WTD group and methotrexate group(P<0.05).The RA model group showed lower relative expression level of typeⅡcollagen mRNA than blank group(P<0.05),while higher relative expression level of p38MAPK mRNA,p38MAPK protein in synovial tissue,serum TNF-αand serum IL-6 than blank group(P<0.05).The methotrexate group,WTD group and WTD+methotrexate group showed higher relative expression level of typeⅡcollagen mRNA than RA model group(P<0.05),while lower relative expression level of p38MAPK mRNA,p38MAPK protein in synovial t

关 键 词：类风湿关节炎 乌头汤 甲氨蝶呤 P38MAPK通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]