Cao Qing-Yong, Wang Pei-Yuan, Wang Qiao-Yun, Zhao Feng, Wang Ke-Zhan, Liu Jun-Qi, Liu Tong-Shen, Wang Yuan-Fang, Liu Yong-Jun
Yantai Affiliated Hospital of Binzhou Medical University, Yantai 264100, China.
School of Pharmacy, Binzhou Medical University, Yantai 264003, China.
Zhongguo Zhong Yao Za Zhi. 2018 Mar;43(5):1034-1041. doi: 10.19540/j.cnki.cjcmm.20180104.004.
The purpose of this experiment is to observe the effects of Tongbi capsule on joint lesions in rabbit with rheumatoid arthritis induced by ovalbumin and explore the mechanism in order to provide reference for clinical application of Tongbi capsule. Rheumatoid arthritis in rabbits was induced by subcutaneous injection of emulsions of ovalbumin and Freund's complete adjuvant and intra articular injection of ovalbumin. After successful modeling, 30 New Zealand rabbits with arthritis were randomly divided into model control group, the high, medium and low dose groups of Tongbi capsule (90, 45, 22.5 mg·kg⁻¹) and prednisone group (5 mg·kg⁻¹). Another six normal rabbits were used as normal control group. After 24 hours of modeling, the rabbits in Tongbi capsule groups received intragastric (i.g.) administrations of Tongbi capsule at 90, 45, 22.5 mg·kg⁻¹·d⁻¹, and the rabbits of prednisone group received i.g. administrations of prednisone at 5 mg·kg⁻¹·d⁻¹ for 2 weeks. The rabbits in normal and model groups received the same volume of distilled water at the same time. The swelling degree of rabbit knee joint and local skin temperature were observed daily. After two weeks of administration, pathological changes of rabbit knee joint were examined by magnetic resonance imaging (MRI); the morphological changes of articular cartilage and synovial membrane were observed by microscope; and the contents of interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-α) in serum were detected by enzyme linked immunosorbent assay (ELISA).The results showed that 24 h after modeling, the knee joints of the rabbits were swollen, with red or dark redlocal skin, and fever, elevated local skin temperature and increased diameters of knee joints. Two weeks after modeling, the swelling of rabbit knee joints was obvious in model group; the joint cavities were filled with purulent fluid; joint synovial membranes were obviously thickened, and even joint cavities were fibrotic and cartilage surfaces showed slight defect; the surface of articular cartilage was obvious fibrosis; synovial epithelial cell proliferation was obvious and accompanied by extensive inflammatory cell infiltration; the levels of IL-1 and TNF-α were significantly higher as compared with those seen in model rabbits (<0.05, <0.01). After 1 and 2 weeks of administration, knee joint diameters and local skin temperatures were smaller or lower than thosein model group (<0.05, <0.01); The lesions of joint cartilage and synovial of all rabbits in each group were less than those in model group; IL-1 and TNF-α levels in serum were also lower than those in model group (<0.05, <0.01). The results reveal that high and medium doses of Tongbi capsule can suppress rheumatoid arthritis induced by ovalbumin in rabbits, reduce joint swelling, inhibit synovial epithelial and fiber hyperplasia and inflammatory cell infiltration, and alleviate articular cartilage damage. The mechanism may be associated with decreasing IL-1 and TNF-α levels in serum.
本实验旨在观察通痹胶囊对卵白蛋白诱导的类风湿性关节炎家兔关节病变的影响,并探讨其作用机制,为通痹胶囊的临床应用提供参考。通过皮下注射卵白蛋白与弗氏完全佐剂的乳剂及关节腔内注射卵白蛋白诱导家兔类风湿性关节炎。造模成功后,将30只患有关节炎的新西兰家兔随机分为模型对照组、通痹胶囊高、中、低剂量组(90、45、22.5 mg·kg⁻¹)和泼尼松组(5 mg·kg⁻¹)。另取6只正常家兔作为正常对照组。造模24小时后,通痹胶囊组家兔分别按90、45、22.5 mg·kg⁻¹·d⁻¹灌胃给予通痹胶囊,泼尼松组家兔按5 mg·kg⁻¹·d⁻¹灌胃给予泼尼松,连续给药2周。正常组和模型组家兔同时给予等体积蒸馏水。每日观察家兔膝关节肿胀程度及局部皮肤温度。给药2周后,采用磁共振成像(MRI)检查家兔膝关节病理变化;通过显微镜观察关节软骨和滑膜的形态变化;采用酶联免疫吸附测定(ELISA)检测血清中白细胞介素-1(IL-1)和肿瘤坏死因子-α(TNF-α)的含量。结果显示,造模24小时后,家兔膝关节肿胀,局部皮肤呈红色或暗红色,发热,局部皮肤温度升高,膝关节直径增大。造模2周后,模型组家兔膝关节肿胀明显;关节腔内充满脓性液体;关节滑膜明显增厚,甚至关节腔纤维化,软骨表面有轻微缺损;关节软骨表面明显纤维化;滑膜上皮细胞增生明显,伴有大量炎性细胞浸润;与模型家兔相比,IL-1和TNF-α水平显著升高(<0.05,<0.01)。给药1周和2周后,膝关节直径和局部皮肤温度均小于模型组(<0.05,<0.01);各组家兔关节软骨和滑膜的病变均少于模型组;血清中IL-1和TNF-α水平也低于模型组(<0.05,<0.01)。结果表明,高、中剂量通痹胶囊可抑制卵白蛋白诱导的家兔类风湿性关节炎,减轻关节肿胀,抑制滑膜上皮和纤维增生以及炎性细胞浸润,减轻关节软骨损伤。其机制可能与降低血清中IL-1和TNF-α水平有关。