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Study of Effects and Mechanism of Strengthening Spleen, Resolving Dampness and Activating Collaterals Herb on Neuro-endocrine-immune Network of Adjuvant Arthritis Rats

Author: WangYuan
Tutor: LiuJian
School: Hubei University of Chinese Medicine
Course: Chinese medical science
Keywords: neuro-endocrine-immune Strengthening spleen, resolving dampness and activating collaterals Rheumatoid arthritis Xinfeng capsule Adjuvant arthritis
CLC: R285.5
Type: PhD thesis
Year: 2010
Downloads: 156
Quote: 0
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Abstract


1.Objective:According to the theory of Tranditional Chinese Medicine (TCM), summing up changes of neuro-endocrine-immune(NEI)network of Rheumatoid Arthritis (RA) and analyzing impacts of Chinese medicine on it.Observing the effect of Strengthening the spleen-Resolving dampness-Promoting collaterals method on neurotransmitters in adjuvant arthritis (AA) rats,and the function of hypothalamus pituitary adrenal (HPA) axis,the changes of vascular neuronpeptide, regulatory T cells and Foxp3 expression, exploring the mechanism of strengthening spleen, resolving dampness and activating collaterals herb on NEI network of AA rats,to enrich the scientific connotation of the treatment of RA based on "Spleen"2.Methods2.1 Theory researchThrough literature review, expounded the relationship between Chinese medicine theory and the NEI network theory,described the changes of RA NEI network and impacts by Chinese medicine. 2.2 Experimental researchEighty-four rats were averagely randomized into normal control(NC) group, model control (MC) group, methotrexate (MTX) treated group, Tripterygium wilfordii polyglycoside Tablet (TPT) treated group, XFC low-dose, middle-dose, high-dose treated groups,12 in each. Except for rats in NC group, the remaining rats in other groups were induced to AA model by intracutaneous injection of 0.1 ml of Freund’s complete adjuvant in the right hindlimb. Serum levels of 5-hydroxytryptamine(5-HT),dopamine(DA),adrenocor ticotro-pic hormone(ACTH),cortisol(CORT),tumor necrosis factor-α(TNF-a),and interleukin-10(IL-10),the levels of endoth-elin-1(ET-1)and calcitionin gene related peptide(CGRP)in serum and brain tissue in all groups were assayed using enzyme linked immunosorbent assay(ELISA).The frequency of CD4+ CD25+ Treg were measured by flow cytometry;The expressions of glucocorticoid receptor(GR) in hypothalamic were measured by immunohistochemistry;The expressions of corticotrophin relea-seeing hormone(CRH) mRNA in hypothalamic,GR mRNA in the cerebral cortex and Foxp3 mRNA in spleen cell were measured by reverse transcription-polymerase chain reaction(RT-PCR);The levels of Foxp3 protein expression in rat thymus tissue were detected by Western blot.3.1 The results of theory researchNEI network theory and TCM theory have very close 1 inks,two-way adjustment and dynamic equilibrium of NEI network were an important manifestation of Yin-Yang theory and the overall concept of TCM, NEI network play an important role in the pathogenesis of RA, while the Spleen in Chinese medicine is an important regulation center of NEI network. The treatment of RA based on "Spleen" can regulate the function of NEI network effectively.3.2 The results of experimental research3.2.1 The treatment based-on "Spleen" can inhibit the neuron-transmitter 5-HT secretion of AA ratsCompared with those in NC group, serum level of 5-HT in AA rats was significantly increased (P<0.05).XFC could signify-cantly decrease serum level of 5-HT,its effect being signify-cantly better than that of MTX and TPT (P<0.05 or P<0.01).No significant difference in DA levels was found among seven groups (P>0.05).3.6.2 The treatment based on "Spleen" can adjust the dys-function of HPA axis in AA ratsCompared with normal group, serum levels of ACTH, CORT and the expression of GRmRNA of cerebral cortex in AA rats were significantly increased (P<0.05,or P<0.01),while the expre-ssion of CRHmRNA in hypothalamic was decreased (P<0.01). Compared with control group,XFC could significantly reduced the levels of ACTH and CORT in serum, and the expression of GRmRNA in cerebral cortex,advanced the expression of CRHmRNA in hypothalamus,better than MTX and TPT group (P<0.05 or P<0.01).3.6.3 The treatment based on "Spleen" can regulate the levels of vascular peptide ET-1,CGRP in AA ratsCompared with the control,ET-1 level increased markedly (P<0.01),and CGRP level decreased markedly (P<0.01);XFC middle and high doses groups can reduce the release of ET-1 and increase the level of CGRP in serum significantly (P<0.01);XFC high and middle dose groups can reduce the release of ET-1 and increase the level of CGRP in brain tissue significantly,better than the control group and XFC low-dose group(P<0.05 or P<0.01),showed a definite dose-effect relationship.3.6.4 The treatment based on "Spleen" can upregulate CD4+CD25+Treg, promote’the expression of Foxp3Compared with normal group,the proportion of CD4+CD25+Treg was significantly decreased (P<0.01),the expression of Foxp3 mRNA in spleen and thymus was significantly lower (P<0.01);Compared with control group,XFC high and middle dose groups can significantly increased CD4+CD25+Treg ratio (P<0.01),increased expression of Foxp3 (P<0.01),significantly better than the MTX group and the XFC low dose group (P<0.05,or P<0.01).3.6.5 Hierarchical cluster analysis of relevant indicators of NEI networkAll indicators is divided into two categories,CORT, TNF-α,5-TH, ACTH, ET in tissue, AI,ET-1, Treg, GRmRNA, joint swelling, and dopamine were elevated in AA rats,they are aggregated priority.In these indicators,the relationships of CORT, TNF-αand 5-TH are most closely; CGRP in tissue and serum, F0Xp3 in thymus and spleen,IL-10 and CRH showed reduced in AA rats,in which 6 indicators CGRP, IL-10 and F0Xp3 are most closely.4.conclusionNEI network theory have very close links with YinYang balance theory and the overall concept of TCM, NEI network play an important role in the pathogenesis of RA.The treatment of strengthening spleen, resolving dampness and activating collaterals can regulated the dysfunction of NEI network. Its effective mechanism relate to the following factors: 4.1 The treatment of strengthening spleen, resolving dampness and activating collaterals can inhibit the secretion of neurotransmitter 5-HT and reduce pain caused by 5-HT,relieve joint symptoms.4.2 The treatment of strengthening spleen, resolving dampness and activating collaterals can maintain the steady-state of HPA axis from the hypothalamus or higher central structures.4.3 The treatment of strengthening spleen, resolving dampness and activating collaterals can effectively adjust the ET-1 and CGRP balance, adjust the synovial angiogenesis systolic and diastolic function, improve joint local vascular dysfunction, play an anti-inflammatory role.4.4 The treatment of strengthening spleen, resolving dampness and activating collaterals by improving CD4+CD25+Treg ratio,enhancing the spleen and thymus Foxp3mRNA and protein expression, inhibit secretion of proinflammatory cytokines,inhibit autoimmune T-cell activation, to achieve the effect of anti-inflammatory and anti-immune.

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