小分子SYK抑制化合物HL131078对胶原诱导性关节炎小鼠模型的治疗作用

发布时间：2018-11-08 20:19

【摘要】：类风湿性关节炎(rheumatoid arthritis,RA)是一种病因尚不明确,以多发性和对称性关节病变为主的非特异性炎症,主要病理变化是关节滑膜的慢性炎症和增生,滑膜边缘增生形成血管翳,关节软骨和软骨下骨被破坏、吸收。RA在治疗方面虽然有了很大的进步,但是目前仍然没有完全根治的治疗办法。治疗RA的传统药物主要有三大类:传统的非甾体类抗炎药、糖皮质激素和生物制剂,这些药物可以改善症状,减缓病情的进展,但不能根治RA,且药物的不良反应也较多。因此,开发安全有效的新型药物将是治疗RA的主要研究方向之一。通过我们前期细胞实验的研究结果表明,小分子SYK抑制化合物HL131078对TNF-α诱导的类风湿关节炎成纤维滑膜细胞的增殖具有良好的抑制作用,且能够抑制成纤维滑膜细胞分泌炎性介质IL-6和MMP-3。目的本课题拟从动物实验水平,观察小分子SYK抑制化合物HL131078对CIA小鼠膝关节滑膜增生、软骨和骨质破坏及分泌IL-6和MMP-3的影响,探讨此化合物对RA的治疗效果,为今后开发治疗RA的新型药物提供依据。研究方法采用CIA小鼠模型,将小鼠分为空白组、模型组、阳性组、实验组(高、低浓度),分别给予灌胃治疗,通过观察小鼠爪子肿胀程度、MIC-CT分析、小鼠膝关节组织石蜡切片HE染色、番红O-固绿染色、免疫组化来评价此小分子化合物对CIA小鼠膝关节滑膜的增生、软骨和骨质的破坏和分泌IL-6、MMP-3的影响。统计学处理数据处理采用SPSS19.0软件,结果以?x±s表示,组间比较采用两样本t检验,P0.05表示差异有统计学差异。结果1、经过灌胃治疗后的实验组小鼠后爪肿胀程度较模型组相比有明显消褪,小鼠后肢平均关节炎指数出现明显下降趋势。2、MIC-CT结果显示,模型组小鼠膝关节骨质破坏严重,实验组小鼠膝关节关节表面未遭到明显侵蚀,骨质仍维持较好完整性。3、在关节组织病理学的结果中,实验组小鼠膝关节组织切片HE染色及番红O-固绿染色可见,滑膜细胞增生不明显,关节腔内仅有少量炎性细胞浸润,膝关节表面软骨侵蚀不严重,与模型组相比,病理学改变明显减轻。4、在免疫组化结果中,实验组小鼠膝关节中IL-6和MMP3的分泌量明显减少。结论小分子SYK抑制化合物HL131078可以有效缓解CIA小鼠膝关节中滑膜的增生,减轻关节表面软骨侵蚀及骨质破坏情况,减少关节腔中IL-6、MMP-3的分泌及炎性细胞的浸润。
[Abstract]:Rheumatoid arthritis (rheumatoid arthritis,RA) is a non-specific inflammation with multiple and symmetrical joint lesions. The main pathological changes are chronic inflammation and hyperplasia of synovial membrane and pannus of synovial edge hyperplasia. Articular cartilage and subchondral bone are destroyed and absorbed. Although RA has made great progress in treatment, there is still no complete cure. There are three main categories of traditional drugs for RA: traditional non-steroidal anti-inflammatory drugs, glucocorticoids and biological agents. These drugs can improve symptoms and slow down the progress of the disease, but can not cure RA, and the adverse drug reactions are more. Therefore, the development of safe and effective new drugs will be one of the main research directions in the treatment of RA. The results of our previous cell experiments showed that the small molecule SYK inhibitor HL131078 had a good inhibitory effect on the proliferation of fibroblasts from rheumatoid arthritis induced by TNF- 伪. It also inhibited the secretion of inflammatory mediators IL-6 and MMP-3. by fibroblast synoviocytes. Objective to observe the effects of small molecule SYK inhibitor HL131078 on synovial hyperplasia of knee joint, cartilage and bone destruction and secretion of IL-6 and MMP-3 in CIA mice at the animal experimental level, and to explore the therapeutic effect of the compound on RA. To provide the basis for the development of new drugs for the treatment of RA in the future. Methods CIA mice were divided into three groups: blank group, model group, positive group, experimental group (high and low concentration), which were treated by gastric perfusion respectively. The degree of paw swelling in mice was observed and analyzed by MIC-CT. The effects of this small compound on synovium proliferation, cartilage and bone destruction and secretion of IL-6,MMP-3 in CIA mice were evaluated by using paraffin sections of mouse knee joint with HE staining, phenanthrene O-solid green staining and immunohistochemistry. SPSS19.0 software was used to process the statistical data. The results were expressed as? X 卤s, and two samples t test were used for the comparison between groups. The difference was statistically significant (P0.05). Results 1. The swelling degree of hind paw in the experimental group was obviously decreased compared with that in the model group, and the average arthritis index of the hind limb of the experimental group was obviously decreased. 2 the results of MIC-CT showed that the swelling degree of the hind paw of the experimental group was significantly decreased than that of the model group. In the model group, the knee joint bone destruction was serious, the surface of the knee joint in the experimental group was not obviously eroded, and the bone still maintained a good integrity. 3 in the results of joint histopathology, In the experimental group, the synovial cell proliferation was not obvious, only a few inflammatory cells infiltrated in the articular cavity, and the cartilage erosion on the surface of the knee joint was not serious, compared with the model group. The pathological changes were obviously alleviated. 4. In the immunohistochemical results, the secretion of IL-6 and MMP3 in the knee joint of the experimental group was significantly decreased. Conclusion small molecule SYK inhibitor HL131078 can effectively relieve synovium proliferation, cartilage erosion and bone destruction, decrease IL-6,MMP-3 secretion and inflammatory cell infiltration in the articular cavity of CIA mice.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R593.22

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