维药刺山柑对类风湿性关节炎滑膜成纤维细胞Th17/Treg免疫失衡调节作用机制的研究

RA is a serious harm to human health, especially in Xinjiang. Capparis Spinosa is the preferred treatment of RA in Uygur Medicine, and has broad prospects.The previous animal experimental study of this project has confirmed that Capparis Spinosa is an effective treatment for RA, which can improve the inflammatory performance of synovium, reduce the release of inflammatory factors, regulate the balance of Th1/Th2 and the expression of IL-17. Based on the interaction between Th17 and Treg cells regulated by Thl and Th2 cells, we proposed a scientific hypothesis that caper citrus can not only regulate Thl/Th2 balance, but also regulate Th17/Treg immune balance. Capparis Spinosa can not only regulate Thl/Th2 balance, but also regulate Th17/Treg immune balance. The aim of this，We'll proceed with the cell experiment，to isolate and culture primary FLS from synovial tissue in vitro, to detect cell proliferation, migration and apoptosis，and to observe the expression of Th17/Treg transcription factor Foxp3, RORrt mRNA and related cytokines IL-17, IL-6, TGF- beta, IL-10, IL-23 and IL-21. To explore the mechanism of Capparis Spinosa on the biological activity, morphology and Th17/Treg immunoregulation of RA-FLS cells, so as to provide reliable experimental basis and theoretical basis for its clinical application, and provide a broader space for the development of traditional ethnic medicine in Xinjiang.

RA严重危害人类健康，在新疆尤其多发。刺山柑是维药治疗RA的首选，有广阔的前景。本项目前期体内实验研究已确证刺山柑是RA有效的治疗手段，能改善滑膜的炎症表现，降低炎性因子的释放，调节Th1/Th2平衡和IL-17的表达。基于Th17和Treg细胞受Thl、Th2细胞的调节，和二者的相互作用关系，我们提出科学假说：刺山柑不仅能调节Thl/Th2平衡，还能调节Th17/Treg免疫平衡。本课题拟从体外实验入手，滑膜组织分离培养原代FLS，检测细胞增殖、迁移能力和凋亡水平，观察Th17/Treg转录因子Foxp3、RORrt mRNA和相关细胞因子IL-17、IL-6、TGF-β、IL-10、IL-23、IL-21的表达。探讨刺山柑对RA-FLS细胞生物学活性、形态学，和Th17/Treg免疫调节的作用机制，为其临床应用提供可靠的实验依据和理论基础，为新疆民族传统医学的发展提供了更广阔的空间。

背景：新疆多发RA，刺山柑是维药治疗RA的首选，有广阔的前景。主要研究内容：检测细胞增殖、迁移能力和凋亡水平，观察Th17/Treg转录因子Foxp3、RORrt mRNA和相关细胞因子IL-17、Ⅱ-6、TGF-β、IL-10、1L-23、IL-21的表达。探讨刺山柑对RA-FLS细胞生物学活性、形态学，和Th17/Treg免疫调节的作用机制。结果：最佳浓度分别为刺山柑粗提取100ng/mL、水苏碱50ng/mL、山萘酚-3-O-芸香糖苷15ng/mL、芦丁30ng/mL和甲氨蝶呤20ng/mL，在最佳浓度下，细胞未出现显著死亡，形态梭形、多角形,具有突起，细胞胞体较大，边界不清晰，大小不均匀，形态正常。与对照细胞组（NC）相比较，水苏碱、山萘酚-3-O-芸香糖苷对RA-FLS细胞均未发现显著作用，而甲氨蝶呤（methotrexate，MT）刺山柑粗提取物（Caper）和芦丁组（Rutin）则具有明显的抑制细胞增殖的作用，在处理后24h-72h，通过MTT法检测细胞的增殖活性，刺山柑粗提取物和芦丁组细胞增殖活性均低于NC组，在72h时，刺山柑粗提取物和芦丁组细胞数量分别为NC组的50.5%和46.5%，在所检测的7个炎性因子中，IL-17和 IL-21在组间无差异，其他5个因子，均存在显著差异性，且在NC中表达高，在处理组中表达显著降低，提示刺山柑发挥抑制RA-FLS细胞增殖、迁移的功能可能是通过降低炎性因子的表达而实现的。意义：可为之后应用于临床提供可靠和理论基础，为新疆民族传统医学的发展提供了更广阔的空间。