摘要
研究獐牙菜苦苷(Swe)对佐剂性关节炎(AA)大鼠的抗炎机制。60只SD大鼠随机分为6组,正常组、模型组、Swe低、中、高剂量组、甲氨蝶呤(MTX)组。AA模型构建方法为大鼠右后趾皮内注射完全弗氏佐剂(CFA) 0.1 m L。各组给药剂量:Swe低、中、高剂量组分别是25,50,100 mg/kg,MTX组为2 mg/kg,空白组与模型组给予等量的生理盐水,第18 d各组按照给药剂量灌胃给药,1次/d,连续给药12 d。酶联免疫试剂盒(ELISA)测定大鼠血清TNF-α、IL-1β水平;荧光定量聚合酶链式反应(RT-PCR)测定大鼠滑膜组织TNF-α、IL-1β的mRNA相对表达量;Western blot法检测大鼠NLRP3、NF-κB通路相关蛋白表达。与正常组比较,模型组大鼠血清TNF-α、IL-1β水平、滑膜组织TNF-α、IL-1β的mRNA相对表达量、p-NF-κB p65、NLRP3、ASC蛋白表达均增多,差异具有统计学意义(P <0.01)。与模型组比较,Swe各剂量组AA大鼠的足体积、关节炎指数评分降低。与模型组比较,Swe中高剂量组TNF-α、IL-1β、TNF-α、IL-1βmRNA的水平降低,差异具有统计学意义(P <0.05)。与模型组比较,Swe中高剂量组滑膜组织p-NF-κB p65、NLRP3、ASC水平明显下调,差异具有统计学意义(P <0.05)。Swe通过抑制NLRP3炎症小体、NF-κB信号通路,发挥对AA大鼠的保护作用。
To study the anti-inflammatory mechanisms of swertiamarin( Swe) on adjuvant-induced arthritis( AA) in rats. There were 60 SD rats randomly divided into 6 groups: the normal group,the AA group,Swe groups( low,medium and high dose)( 25,50,100 mg/kg),the methotrexate( MTX) group( 2 mg/kg). The AA model was induced by intradermal injection of complete Freund’s adjuvant( CFA) for 0. 1 m L into the right hind paw of the rats. On the 18 th day,each rat received drug by intragastric administration for 12 days. The dosages for each group: corresponding doses for the low,medium and high dose Swe groups were 20 mg/kg,10 mg/kg,50 mg/kg. The dose for MTX group was 2 mg/kg. The normal group and the model group were given equal volume of saline. The serum TNF-α and IL-1β levels in rats were determined by Enzyme Linked Immunosorbent Assay( ELISA) Kits. Realtime fluorescent quantitative polymerase chain reaction( RT-PCR) was used to determine the expression of TNF-α,IL-1β mRNA levels in the synovial tissues of rats. The expression of proteins involved in NLRP3 inflammasome,NF-κB signaling pathways were measured by western blot. Compared with the normal group,the serum level of TNF-α and IL-1β increased significantly in rats,the relative expression of TNF-α and IL-1β mRNA was significantly increased and the expression of p-NF-κB p65、NLRP3 and ASC proteins was significantly increased( P < 0. 01) in synovial tissue after modeling. Compared with the model group,the paw volume and the arthritis index scores of AA rats were decreased in each dose Swe group. Compared with the model group,the level of serum TNF-α,IL-1β,TNF-α mRNA,IL-1β mRNA and p-NF-κB p65,NLRP3,ASC levels were significantly reduced( P < 0. 05) in the medium and high dose Swe groups. Therapeutic effects of swertiamarin on adjuvant-induced arthritis rats were by inhibiting NLRP3 inflammasome and NF-κB signaling pathways.
作者
王新强
吴良邦
章月红
顾增辉
WANG Xin-qiang;WU Liang-bang;ZHANG Yue-hong;GU Zeng-hui(Department of Orthopedics,903 Hospital,Joint Logistics Support Force,Chinese People’s Liberation Army,Hang-zhou 310004,China)
出处
《药物生物技术》
CAS
2021年第2期123-128,共6页
Pharmaceutical Biotechnology
基金
浙江省医药卫生科技计划项目(No.2019329727)。
