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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 observe the effects of alcohol extracts from Nanzha rattan on synovial tissue proliferation and cartilage erosion and degradation of rheumatoid arthritis (RA) and to explore the mechanism for RA drug discovery. Methods The synovial synovium of RA patients and normal articular cartilage were transplanted into NOD/SCID mice to establish the human RA synovial-cartilage-NOD/SCID mouse chimera model (NOD/SCID-HuRAg). Distilled water was given 4 weeks later. Nanzha rattan alcohol extract (30mg/d) and leflunomide (500μg/d) were intragastrically administered once a day for 4 weeks. The histological scores of synovial hyperplasia, cartilage erosion and cartilage cell degradation around the grafts were evaluated, and serum TNF-α content was measured by radioimmunoassay. In situ hybridization was used to observe the expression of TNF-α mRNA in the synovial membrane. The TUNEL method was used to observe the degree of apoptosis in synovial membrane. The results were analyzed by automatic image analysis system. Results The synovial membrane and cartilage grew well in SCID mice. The growth of synovial membrane was significantly reduced by the alcohol extracts of Nepentena rapa and leflunomide (2.00±0.76, 2.25±0.89 vs 3.63±0.52, respectively) and cartilage erosion (1.69). ± 0.80, 2.00 ± 1.36 vs 3.75 ± 0.53) and cartilage degradation (1.88 ± 0.83, 2.13 ± 0.83 vs. 3.63 ± 0.74) scores, and significantly reduced serum TNF-α levels (0.84 ± 0.09, 0.83 ± 0.12 vs 0. 99 ± 0.11, ng/ml). Both drugs significantly increased the apoptosis of synovial cells, and the ethanol extract from Nanzip can significantly down-regulate the expression of TNF-α in synovial tissue. Conclusion The ethanol extract from Nanzip can inhibit the synovial hyperplasia of SCID-HuRAg model, reduce the cartilage erosion of synovium and chondrocyte-mediated cartilage degradation. The mechanism of its action includes inhibiting the production of TNF-α in RA synovial tissue. Promote synovial cell apoptosis. The effect of the ethanol extract of Nanshang was similar to that of leflunomide, but it was stronger than leflunomide in inhibiting the expression of TNF-α in RA synovium.