Hu Yiping, Li Jinchao, Qin Ling, Cheng Wenxiang, Lai Yuxiao, Yue Ye, Ren Peigen, Pan Xiaohua, Zhang Peng
Department of Orthopaedics, Bao'an People's Hospital, Shenzhen, People's Republic of China.
1068 Xueyuan Avenue, Shenzhen University Town, Shenzhen, 518055, China.
Curr Pharm Des. 2016;22(46):6975-6981. doi: 10.2174/1381612822666161025150403.
This work aimed to evaluate the effects of genistein treatment in Collagen Induced Arthritis (CIA) mouse model.
CIA was elicited in DBA/1 Mice by an intradermal injection of 100 μL of an emulsion of bovine type II collagen (CII) in isovolumic incomplete Freund's adjuvant (IFA) at the base of the tail. Twenty-one days later, a second injection of CII in IFA was administered at the base of the tail. After the symptoms of arthritis showed in mouse model, we divided animals into two groups according to their clinical symptom scores. The treatment group was intraperitoneally injected daily with genistein (based on the pre-experiment data and literature reported, 5 mg/kg dose was selected and tested) for 12 days, while the control group was injected with phosphate buffered saline. Inflammatory cytokines titer, radiological, and histological observations were completed at different time's points after treatment. CT analysis was conducted 3 months after the treatment to observe the articular structures. Immunohistochemical analysis was performed to investigate the expression and distribution of VEGF in joint tissues.
Genistein suppressed the expressions of IL-1β, IL-6 and TNF-α in the serum. Radiological results showed that bone degradation was inhibited by the treatment. Moreover, hematoxylin and eosin staining showed that the degree of inflammation was relieved. In the cartilage area, TRAP stain-positive cells were detected, which was notably reduced in the treatment group compared to the control group. Micro-CT 3D images clearly exhibited that the joint adhered and structures destroyed in the control group with less destruction in the treatment group. Furthermore, genistein suppressed VEGF expression, and blocked angiogenesis in the synovial tissue.
Our work provides further data regarding the effects of genistein as a potential treatment drug for RA, as well as the role of genistein in the anti-inflammatory pathway in RA therapy.
本研究旨在评估染料木黄酮治疗对胶原诱导性关节炎（CIA）小鼠模型的影响。
通过在DBA/1小鼠尾部基部皮内注射100 μL牛II型胶原（CII）与等体积不完全弗氏佐剂（IFA）的乳剂诱发CIA。21天后，在尾部基部再次注射CII与IFA的乳剂。在小鼠模型出现关节炎症状后，根据临床症状评分将动物分为两组。治疗组每天腹腔注射染料木黄酮（根据前期实验数据和文献报道，选择并测试5 mg/kg剂量），持续12天，而对照组注射磷酸盐缓冲盐水。在治疗后的不同时间点完成炎症细胞因子滴度、放射学和组织学观察。治疗3个月后进行CT分析以观察关节结构。进行免疫组织化学分析以研究VEGF在关节组织中的表达和分布。
染料木黄酮抑制血清中IL-1β、IL-6和TNF-α的表达。放射学结果显示治疗可抑制骨破坏。此外，苏木精-伊红染色显示炎症程度减轻。在软骨区域，检测到抗酒石酸酸性磷酸酶（TRAP）染色阳性细胞，与对照组相比，治疗组显著减少。显微CT三维图像清楚地显示对照组关节粘连且结构破坏，而治疗组破坏较少。此外，染料木黄酮抑制VEGF表达，并阻断滑膜组织中的血管生成。
我们的研究提供了关于染料木黄酮作为类风湿关节炎潜在治疗药物的作用的进一步数据，以及染料木黄酮在类风湿关节炎治疗抗炎途径中的作用。
