痰瘀型类风湿性关节炎与淋巴关系的基础研究

Rheumatoid arthritis (RA) a systemic autoimmune disease with severe joint lesions. Phlegm and blood stasis play an important part in the RA process, and their characteristic also leads to complexity of RA. There is no effective treatment on RA. Therefore, it is urgent to in-depth study of the biological characteristics of the onset of phlegm and blood stasis. .Our study group observed that lymphangiogenesis in RA and some other inflammatory joint, and proved that promoting lymphangiogenesis and lymphatic function are targets for the treatment of such diseases, and Arctium soup usually achieved significant results..Based on the above ideas and research base, we will further use transgenic mice, using ICG-near infrared, MRI, molecular biology and technology to in-depth study of the regulation effect of lymphatic function on inflammatory arthritis, and explore the effect of Jiawei Arctium soup and its effective component on the regulation of Notch signaling pathway, VEGF-C expression and lymphatic function changes. .Based on the clinical research of phlegm and blood stasis type RA patients, we will explore the relationship between the ZhuTanQuYu law and lymphatic function, discuss the biological basis of the traditional Chinese medicine (TCM) “Phlegm and blood stasis” theory between inflammation and lymphoid hyperplasia and functions, guide the increase of clinical levels and the development of the “phlegm and blood stasis” theory of TCM .

类风湿关节炎（RA）是以关节病变为主的全身慢性自身免疫性疾病，痰瘀在该病的发生发展中具有重要作用，其粘滞凝涩的特性决定了RA复杂难愈的特点，临床上也缺乏有效的治疗药物。因此，急需深入研究RA痰瘀证发生的生物学基础。.本项目组前期研究观察到RA等炎症关节局部均有淋巴组织的增生，并证明了促进淋巴增生和功能是减轻慢性炎症反应、治疗该类疾病的靶点，临床上采用加味牛蒡子汤逐痰祛瘀治疗RA也取得明显疗效。.基于上述思路和研究基础，本项目将通过临床试验和应用已建立的TNF-Tg炎性关节炎小鼠模型和淋巴体内成像技术，进一步研究痰瘀型关节炎的发病机理、病理变化与淋巴功能之间的相关性，寻找新的治疗靶点；并进一步研究“加味牛蒡子汤”调控Notch 信号通路、血管内皮生长因子C表达及淋巴管功能变化，探讨中医“逐痰祛瘀”与淋巴功能之间的关系，发展中医痰瘀理论。

在本重点项目的资助下，我们完善了类风湿关节炎（RA）淋巴回流功能检测技术平台：1. 建立吲哚菁绿—近红外(Indocyanine Green—Near Infrared，ICG-NIR)大动物绵羊踝、膝关节淋巴回流功能检测技术；2. 建立了转基因斑马鱼淋巴管生成中药筛选技术；3. 建立胶原诱导关节炎（Collagen induced arthritis，CIA）小鼠踝、膝关节滑液体积与血液流动多普勒超声检测技术；进一步阐释了RA淋巴回流功能障碍的分子机制，证明了“淋巴系统是参与RA痰瘀病理变化的关键环节”：1. 发现TNF直接刺激淋巴管平滑肌细胞 (lymphatic smooth muscle cells, LSMCs) 标记基因减少和凋亡；2. 发现慢性RA炎症刺激淋巴管内皮细胞 (Lymphatic endothelial cells, LECs) 产生NO损伤 LSMCs，造成淋巴回流功能障碍；3. 发现M1巨噬细胞引起LECs炎症反应，造成淋巴回流功能障碍，加重关节炎症发生的程度。阐释加味牛蒡子汤调控淋巴回流功能和淋巴管生成治疗痰瘀型RA的疗效和机制：1. 加味牛蒡子汤调控淋巴回流功能和淋巴管生成，发挥治疗痰瘀型RA的独到作用；2. 三七总皂苷促进LECs表达VEGF-C促进淋巴管生成；3.人参皂苷Rg1通过刺激LECs表达VEGF-C，促进淋巴回流功能，减轻CIA小鼠关节炎症；4. 三七皂苷R1通过刺激LECs表达VEGF-C，促进淋巴回流功能，减轻CIA小鼠关节炎症；5. 龙胆苦苷抑制巨噬细胞向M1方向极化；6. 蛇床子素抑制TNFα诱导的滑膜细胞炎症因子表达和降解酶的表达，减轻CIA小鼠关节炎症状；7. 紫云英苷抑制TNFα诱导的滑膜细胞炎症因子表达和降解酶的表达，减轻CIA小鼠关节炎症状。完成了加味牛蒡子汤颗粒联合甲氨蝶呤治疗RA多中心、随机、双盲、安慰剂对照试验，结果显示加味牛蒡子汤颗粒改善RA患者的关节疼痛，肿胀等临床症状。