基于VEGF/ VEGFR-2、PLC-γ/ MAPK 信号转导通路探讨痰瘀致痹与类风湿关节炎血管增生的关系

Arthrosynovitis of joints is a significant pathology of Rheumatoid Arthritis(RA), which bases on abnormal angiogenesis. Vascular endothelial growth factor(VEGF) and its signal pathways regulate the development of angiogenesis. According to the comprehendsion of traditionnal Chinese medicine physician about RA,which was based on rheumatic arthralgia induced by phlegm and blood stasis, we concluded that phlegm conglutinating with blood stasis was a key pathogeny throughout the progression of RA. We also found that the satisfactory therapeutic effect of RA by resolving phlegm and dredging collaterals. In the past research, we found the therapy of resolving phlegm and dredging collaterals could alleviate the inflammation of joints, and downregulate the expressions of VEGF in serum of human and rats. Therefore, We conjectured that the therapy way of resolving phlegm and dredging collaterals would depress vascular proliferation through reglating VEGF, VEGF receptor and endothelial cells' signaling pathways. The scores of phlegm syndrome and blood stasis syndromn will be observed as our Clinical study of indexs.The human serum, human synovial fluid of joint, synovium of collagen-induced arthritis(CIA) rat's joint and human umbilical vein endothelial cells (HUVEC) will be chosen as objects to investigate the inhibiting effect of VEGF, VEGFR-2 and angiogenesis, as well as the VEGF/VEGFR-2, PLC-γ/MAPK Signal Pathways, respectively, by resolving phlegm and dredging collaterals. From these study, we will identify the key mechanism of resolving phlegm and dredging collaterals therapy to RA, explore the scientific connotation of phlegm and blood stasis and prove a new basis for investigational new TCM medicine.

类风湿关节炎(RA)的基本病理表现为关节滑膜炎，血管异常增生是滑膜炎形成的基础，而血管内皮生长因子（VEGF）及效应细胞的信号转导过程决定着血管增生的发展方向。根据历代医家对痰、瘀致痹的认识，我们总结出痰浊、瘀血作为共性的病理变化贯穿于RA的发病过程，并以化痰通络法治疗RA取得了满意的疗效。前期，我们发现化痰通络法可降低人、大鼠血清VEGF 的表达，并减轻关节炎症，推测化痰通络法可能通过抑制血管增生治疗RA。 本课题以痰证、瘀证积分为主要临床研究指标，并以人血清、关节滑液、大鼠的关节滑膜及人脐静脉内皮细胞为实验研究对象，检测化痰通络法对VEGF、VEGF受体及微血管增生的作用，并探讨化痰通络法对VEGF 相关内皮细胞主要的信号转导通路VEGF/VEGFR-2、PLC-γ/MAPK 影响，从而明确化痰通络法的治疗机制，探讨痰瘀致痹的科学内涵，为开发治疗RA 的新药提供依据。

类风湿关节炎(RA)的基本病理表现为关节滑膜炎，血管异常增生是滑膜炎形成的基础，而血管内皮生长因子（VEGF）及效应细胞的信号转导过程决定着血管增生的发展方向。根据历代医家对痰、瘀致痹的认识，我们总结出痰浊、瘀血作为共性的病理变化贯穿于RA的发病过程，并以化痰通络法治疗RA取得了满意的疗效。本课题以痰证、瘀证积分为主要临床研究指标，并以人血清、关节滑液、 大鼠的关节滑膜及人脐静脉内皮细胞为实验研究对象，检测化痰通络法对VEGF、VEGF受体及微血管增生的作用，并探讨化痰通络法对VEGF 相关内皮细胞主要的信号转导通路VEGF/VEGFR-2、PLC-γ/MAPK 影响。化痰通络方含药血清对VEGF诱导的HUVEC细胞增殖具有抑制作用。设计的siRNA 可针对性的敲降HUVEC 中VEGFR-2/PLCG1/MAPK1表达，雷公藤多苷组抑制VEGF诱导的HUVEC细胞增殖依赖于VEGFR2和PLCG1表达，而化痰通络方对VEGF刺激的HUVEC细胞的增殖的抑制效应则不仅仅依赖VEGF/VEGFR2/PLCG1/MAPK1信号通路。从而明确化痰通络法的治疗机制，探讨痰瘀致痹的科学内涵，为开发治疗RA 的新药提供依据。