杜仲三萜、环烯醚萜类成分对胶原诱导性关节炎大鼠的影响

Effects of triterpenoid and iridoid of Eucommiae Cortex on collagen-induced arthritis in rats

杜仲醇提物乙酸乙酯部位能有效改善胶原诱导性关节炎大鼠(collagen-induced arthritis,CIA)的关节炎症以及骨破坏情况,对类风湿关节炎有潜在的治疗作用。杜仲三萜及环烯醚萜是从杜仲醇提物乙酸乙酯部位进一步分离得出的萜类物质,其对CIA大鼠的关节炎症和骨侵蚀是否具有抑制作用尚不明确。该研究通过诱导CIA大鼠模型,观察杜仲2种萜类物质及其两者合用对大鼠关节炎的影响,进一步明确传统中药杜仲抗关节炎的物质基础,同时在给药后2、4周分别取材,观察CIA大鼠关节炎的不同阶段病理改变。对于模型对照组大鼠,随着病程的延长,足容积、关节炎评分与组织病理学病变加重,与模型对照组相比,各时期萜类给药组均可以明显降低大鼠足容积与关节炎的评分以及改善关节病变与软骨破坏。此外,各萜类给药组可以显著下调各时期脾脏中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-17(interleukin-17,IL-17)、白细胞介素-23(interleukin-23,IL-23)的mRNA表达量。在体外,杜仲总萜、三萜在质量浓度为160、320μg·mL^(-1)可以显著抑制人类风湿关节炎成纤维样滑膜细胞(human fibroblast-like synoviocyte cells-RA,HFLS-RA)增殖,在10~80μg·mL^(-1)可以显著抑制脂多糖(lipopolysaccharide,LPS)诱导的RAW264.7释放一氧化氮(nitric oxide,NO);杜仲环烯醚萜对HFLS-RA细胞生长无影响,但在质量浓度为40、80μg·mL^(-1)可以显著抑制RAW264.7细胞NO释放量。综上结果表明杜仲萜类成分对于类风湿关节炎的治疗具有较大的潜力。

The ethyl acetate fraction of ethanol extract of Eucommiae Cortex can effectively inhibit joint inflammation and bone destruction in rats with collagen-induced arthritis(CIA)and has a potential therapeutic effect on rheumatoid arthritis.The triterpenoid(EU-Tid)and iridoid(EU-Idd)of Eucommiae Cortex are derivatives isolated from the ethyl acetate fraction of the ethanol extract of Eucommiae Cortex,and it is not clear whether they have inhibitory effects on joint inflammation and bone erosion in CIA rats.Therefore,based on the CIA model,the effects of EU-Tid,EU-Idd,and their combination(EU-TP)on arthritis in rats were observed,and the material basis of Eucommiae Cortex against arthritis was further clarified.The samples were collected two and four weeks after administration to observe the pathological changes in different stages of arthritis in CIA rats.For the rats in the model control group,with the prolongation of the disease course,the paw volume and arthritis score increased and histopathological lesions aggravated.Compared with the model control group,the drug administration groups showed reduced paw volumes and arthritis scores,and improved joint lesions and cartilage destruction.Additionally,the mRNA expression levels of tumor necrosis factor-α(TNF-α),interleukin-17(IL-17),and interleukin-23(IL-23)in the spleen were down-regulated in the drug administration groups.EU-TP and EU-Tid at concentrations of 160 and 320μg·mL^(-1) could significantly inhibit the proliferation of human fibroblast-like synoviocytes-RA(HFLS-RA)and nitric oxide(NO)release in the supernatant of RAW264.7 cells induced by lipopolysaccharide(LPS)at the concentration range of 10-80μg·mL^(-1) in vitro.EU-Idd had no effect on the proliferation of HFLS-RA but could reduce the NO release at concentrations of 40 and 80μg·mL^(-1).The results indicated that the terpenoids of Eucommiae Cortex had great potential in the treatment of rheumatoid arthritis.