Study On The Treatment Of Rheumatoid Arthritis By Jia-weiniu-bang-zi-tang And Calycosin To Reduce Inflammation Of Lymphatic Endothelial Cells

Objective: To study the anti-inflammatory and immune effects and its pharmacological mechanism of Jia-Wei-Niu-Bang-Zi-Tang and active ingredient calycosin in inhibiting LECs inflammation induced by macrophage inflammation and regulating structure and function fo lymphatic system in the prevention and treatment of rheumatoid arthritis.Methods: An animal model of TNF-α overexpression in spontaneous rheumatoid arthritis(TNF-Tg mice)was established in vivo,and the mice were treated with Jia-WeiNiu-Bang-Zi-Tang or calycosin by ingastric administration.The changes of body weight,RA symptom score and behavior of heat pain threshold of the mice after treatment were observed.The effects of drugs on RA inflammation by regulating lymphatic system were evaluated from the aspects of three-dimensional imaging volume of popliteal lymph nodes,function of lymphatic system and pathological morphological changes of ankle joint.The primary macrophages were isolated in vitro and cultured with 20ng/m L MCSF,and lymphatic endothelial cell lines were cultured.Flow cytometry,RT-q PCR,Western blot,immunofluorescence and other methods were used to study the inflammatory effect of calycosin on macrophages stimulated by TNF-α and its effect on inflammatory response of lymphatic endothelial cells,so as to explore the pharmacological mechanism of its prevention and treatment of RA.Results: The experimental results showed that Jia-Wei-Niu-Bang-Zi-Tang could reduce the number of M1-type macrophages in TNF-Tg mice,promote lymphatic angiogenesis,reduce the inflammatory immune response of m1-type macrophage aggregation in the inflammatory site of join,and promote cartilage and bone repair in ankle joint.Calycosin can improve the lymphatic reflux function of TNF-Tg mice,promote the regeneration of lymph-vessel,improve the inflammatory local microcirculation,and reduce RA.Calycosin inhibited macrophage inflammation stimulated by TNF-α(5ng/m L),inhibited the expression of JNK protein in NF-κB signaling pathway p65,upstream signaling molecule IKKα/β and MAPK signaling pathway,and inhibited the phosphorylation of p65 protein into the nucleus.Macrophage inflammation stimulated by TNF-α(5ng/m L)can induce inflammation of lymphatic endothelial cells and form an inflammatory cycle,but the inflammatory stimulation of macrophage inflammation on lymphatic endothelial cells was weakened after 50μM calycosin intervention.Conclusion: Jia-Wei-Niu-Bang-Zi-Tang can repair lymphatic structure in RA state,and its active ingredient Calycosin can reduce macrophages inflammation by inhibiting NF-κB and JNK signaling pathway of macrophages,reduce its pro-inflammatory effect on lymphatic endothelial cells,improve lymphatic reflux function,and reduce joint inflammation in TNF-Tg mice.These results further elucidate the pharmacological mechanism of Jia-Wei-Niu-Bang-Zi-Tang and its active ingredient in regulating lymphatic function in the treatment of rheumatoid arthritis,and provide preclinical evidence for its clinical application.Figure [43] Table [2] Reference [87]...