白术多糖对类风湿性关节炎大鼠的抗炎作用及TLR4/NF-κB信号通路的影响

Effect of polysaccharide of atractylodes macrocephala koidzon anti-inflammatory effects and TPLR4/NF-κB signaling pathwayin rats with rheumatoid arthritis

目的探讨白术多糖(PAMK)对类风湿性关节炎(RA)大鼠的抗炎作用及其对Toll样受体4/核因子κB(TLR4/NF-κB)信号通路的影响。方法采用Ⅱ型胶原诱导建立RA大鼠模型,造模成功后将大鼠分为模型组、阳性药物组(雷公藤多苷片)、PAMK低、高剂量组,同时设置正常对照组,每组12只。给予雷公藤多苷片或PAMK灌胃给药,1次/d,连续4周。测量大鼠体质量、足趾容积和关节炎指数;计算胸腺指数和脾脏指数;HE染色观察踝关节滑膜组织病理学变化;ELISA法检测血清及踝关节滑膜组织中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)及肿瘤坏死因子-α(TNF-α)水平;Western blot检测踝关节滑膜组织中总蛋白Toll样受体4(TLR4)、髓样分化因子88(MyD88)、磷酸化核因子κB p65[p-NF-κB p65(Ser536)]及核蛋白NF-κB p65表达水平。结果与正常对照组比较,模型组大鼠可见滑膜增生、层次不清等病理学改变,体质量降低(P<0.05),足趾容积、关节炎指数以及胸腺、脾脏指数升高(P<0.05),血清及踝关节滑膜组织中IL-1β、IL-6和TNF-α水平以及踝关节滑膜组织中TLR4、MyD88、p-NF-κB p65和NF-κB p65蛋白表达水平升高(P<0.05)。与模型组比较,阳性药物组和PAMK高剂量组大鼠踝关节滑膜组织病理学改变明显改善,体质量升高(P<0.05),足趾容积、关节炎指数以及胸腺、脾脏指数降低(P<0.05),血清及踝关节滑膜组织中IL-1β、IL-6和TNF-α水平以及踝关节滑膜组织中TLR4、MyD88、p-NF-κB p65和NF-κB p65蛋白表达水平降低(P<0.05),而PAMK低剂量组大鼠以上各指标与模型组比较差异均无统计学意义(P>0.05)。结论PAMK可减轻机体炎症反应,改善RA,其机制可能与抑制TLR4/NF-κB信号通路的活化有关。

Objective To investigate the anti-inflammatory effect of polysaccharide from atractylodes atractylodes(PAMK)on rheumatoid arthritis(RA)rats and its effect on Toll like receptor 4/nuclear factor kappa-B(TLR4/NF-κB)signaling pathway.Methods RA rat model was induced by typeⅡcollagen,after successful modeling,the rats were divided into model group,positive drug group(Tripterygium wilfordii polyglycoside tablet),PAMK low-dose and high-dose groups,normal control group was also set,with 12 rats in each group.Tripterygium wilfordii polyglycoside tablets or Atractylodes macrocephala polysaccharide were administered intragastric administration,once a day for 4 weeks.The body weight,paw swelling rate and arthritis index of rats were measured;the thymus index and spleen index were calculated;HE staining was used to observe the histopathological changes of synovial membrane of ankle joint;The levels of interleukin-1β(IL-1β),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in serum and synovial tissue of ankle were determined by ELISA;The expression levels of total protein TLR4,MyD88,phosphorylated NF-κB p65(Ser536)and nuclear protein NF-κB p65 in synovial tissue of ankle were detected by Western blot.Results Compared with the normal control group,synovial hyperplasia and unclear layers were observed in the model group,body weight decreased(P<0.05),paw swelling rate,arthritis index,thymus index and spleen index increased(P<0.05),the levels of IL-1β,IL-6 and TNF-αin serum and synovial tissue of ankle,and the protein expressions of TLR4,MyD88,p-NF-κB p65 and NF-κB p65 in synovial tissue of ankle increased(P<0.05).Compared with model group,the histomathological changes of synovial of ankle joint in positive drug group and PAMK high-dose group were significantly improved,body weight increased(P<0.05),paw swelling rate,arthritis index,thymus index and spleen index decreased(P<0.05),the levels of IL-1β,IL-6 and TNF-αin serum and synovial tissue of ankle,and the protein expressions of TLR4,MyD88,p-NF-κB p65 and NF-κB p65 in synovial tissue of ankle decreased(P<0.05),however,there were no significant differences in the above indexes between PAMK low-dose group and model group(P>0.05).Conclusion PAMK can reduce the inflammatory response and improve RA,and the mechanism may be related to the inhibition of TLR4/NF-κB signaling pathway activation.