南蛇藤醇提物对类风湿关节炎滑膜增生和软骨侵蚀及降解的抑制作用

Methanol extract of Celastrus orbiculatu suppresses synovial hyperplasia and cartilage erosion and degradation in rheumatoid arthritis

目的观察南蛇藤醇提物对类风湿类关节炎(RA)滑膜组织增生和软骨侵蚀及降解的作用并探讨其机制,为RA的新药研究提供依据。方法将RA患者的关节滑膜和正常关节软骨移植到NOD/SCID小鼠体内建立人RA滑膜-软骨-NOD/SCID鼠嵌合体模型(NOD/SCID-HuRAg),4周后分别给予蒸馏水、南蛇藤醇提物(30mg/d)及来氟米特(500μg/d)灌胃,每日1次,持续4周。评价移植物中的滑膜增生、滑膜细胞对软骨的侵蚀和软骨细胞周围软骨降解的组织学积分,放免法检测血清TNF-α含量。原位杂交法观察滑膜的TNF-αmRNA表达,TUNEL法观察滑膜的细胞凋亡程度,自动图像分析系统分析结果。结果滑膜和软骨在SCID鼠体内生长良好,南蛇藤醇提物及来氟米特能显著降低滑膜增生(分别为2.00±0.76,2.25±0.89vs3.63±0.52)、软骨侵蚀(1.69±0.80,2.00±1.36vs3.75±0.53)和软骨降解(1.88±0.83,2.13±0.83vs3.63±0.74)的积分,并显著降低血清TNF-α含量(0.84±0.09,0.83±0.12vs0.99±0.11,ng/ml)。二种药物均显著增加了滑膜细胞的凋亡,南蛇藤醇提物还显著下调了滑膜组织中TNF-α的表达水平。结论南蛇藤醇提物能抑制SCID-HuRAg模型的滑膜增生,减轻滑膜的软骨侵蚀和软骨细胞介导的软骨降解,其作用的机制包括抑制RA滑膜组织的TNF-α的产生和促进滑膜细胞凋亡。南蛇藤醇提物的作用效果与来氟米特相似,但在抑制RA滑膜TNF-α表达方面南蛇藤醇提物强于来氟米特。

Objective To investigate the effects of methanol extract of Celastrus orbiculatu （MECO） on synovial hyperplasia and cartilage erosion and degradation in rheumatoid arthritis （RA）, and explore the possible mechanisms to provide clues for new drug development for RA treatment. Methods The articular synovium from patients with RA and normal articular cartilage were co-implanted into the back of severe combined irnmunodeficient （SCID）mice to establish the chimeric model SCID- HuRAg. Four weeks later, the mice were given MECO intragastrically at 30 mg/day, leflunomide at 500 μg/day or distilled water, respectively, for 4 consecutive weeks. After completion of the treatments, the histological scores of the grafts for synovial hyperplasia, cartilage invasion by synoviocyte and cartilage degradation around the chondrocytes were evaluated, and serum level of tumor necrosis factor-α （TNF-α） was measured with radioimmunoassay. The expression of TNF-α mRNA and the cell apoptosis in the synovium were detected with in situ hybridization （ISH） and TUNEL, respectively, and the results were analyzed with the image analysis system. Results The grafts survived in the mice till the end of experiment. MECO and leflunomide, in comparison with distilled water, significantly lowered the scores for synovial hyperlasia （2.00±0.76 and 2.25±0.89 vs 3.63±0.52）, cartilage erosion （1.69±0.80 and 2.00±1.36 vs 3.75±0.53）, cartilage degradation （1.88±0.83 and 2.13±0.83 vs 3.63±0.74） and serum TNF-α level （0.84±0.09 and 0.83±0.12 vs 0.99±0.11 ng/ml）. Cell apoptosis of the synovium increased significantly with MECO and leflunomide treatments, but the expression of TNF-α mRNA in the synovium decreased significantly in MECO group. Conclusion MECO can effectively suppress synovial hyperplasia and cartilage erosion and degradation SCID-HuRAg mice by reducing TNF-α production in the synovium and promoting synovial apoptosis. MECO can be comparable with leflunomide in their effect, but the former is more effective in suppressing TNF-α expression in the synovium.