| Chapter1IntroductionRheumatoid Arthritis is a autoimmune diseases that results in a chronic, systemic inflammatory disorder that presents with joint inflammation and synovitis. Chinese Medicine name RA as WangBi and "Wang" is referred as weakening of hind limb and body, disability to walk and damage to joint and bone. Traditional Chinese Medicine is effective in the treatment of RA. Previous studies also showed that Bu Shen Tong Du Capsule (BS) is effective in treating RA and osteoarthritis. Therefore, our study aimed to investigate the therapeutic mechanism of Bu Shen Tong Du Capsule on RA.Chapter2Literature ReviewReview1The literature review of RA in Chinese Medicine mainly introduced the definition, etiology, pathology and treatment progress of RA in Chinese Medicine.Review2The literature review of RA in Western Medicine mainly introduced the definition, etiology and treatment progress of RA in Western Medicine and provided the pathology of specific immunity and osteoclasia in RAReview3The literature review of animal model used in RA mainly introduced several commonly used animal model involved in the study of etiology, pathology and drug mechanism of RA.Review4The literature review of Chinese Medicine mechanism in treating RA mainly introduced the action mechanism of single herbs or herbal remedy and suggested immune modulation and bone protection as the research key.Chapter3Laboratory ResearchResearch1The Therapeutic Effect of BS on CIA rat modelAim:To investigate the effect of BS on CIA rat modelMethod:42female SD rats were randomly assigned to six groups, normal control group(C), model group(M), low dose of BS group(BSL), medium dose group(BSM), high dose group (BSH) and tripterygium glycosides group (TG). Arthritis would be induced on day1and7by injection of incomplete freund’s adjuvant and bovine type Ⅱ collagen. After induction of arthritis on day1, the weight,foot thickness and arthritis scores were measured every2to3days. From Day13, the rat was administrated with BSL120mg/d/kg, BSM240mg/d/kg, BSH480mg/d/kg, TG24mg/d/kg and saline for18days. Results:BS could significantly reduced foot thickness and arthritis scores of CIA rat(p<0.05). Also, BS is effective in relieving the effect of arthritis on weight.Conclusion:BS could significantly relieve arthritis. Research2The Modulation Effect of BS on Thl and Th2cells of CIA rat modelAim:To investigate the effect of BS on spleen pathology, Thl cell and Th2cell related cytokines of CIA rat modelMethod:42female SD rats were randomly assigned to six groups, C group, M group, BSL group, BSM group, BSH group and TG group. Arthritis would be induced on day1. From Day13, the rat was administrated with BSL120mg/d/kg, BSM240mg/d/kg, BSH480mg/d/kg, TG24mg/d/kg and saline for18days. At day31, rats were sacrificed to perform(1)HE stain of spleen;(2)Flow cytometry to analyze peripheral percentage of Thl and Th2cell;(3)RT-PCR to analyze RNA quantity of IFN-γ and IL-4;(4)ELISA to detect serum IFN-y, TNF-a,IL-4and IL-10.Results:(1)BS could relieve white pulp hyperplasia of spleen.(2)BS could down regulate Th1%and up regulate Th2%(p<0.01) to restore balance of Th cell.(3)BS could down regulate IFN-Y RNA presentation and up regulate IL-4RNA presentation.(4) Serum levels of IFN-γ、TNF-α presented downward trend and IL-4and IL-10presented upward trend while the dose of BS was raised.Conclusion:BS could suppress humoral immune response, restore the balance of Th cells and suppress production and action of inflammatory cytokines to relieve joint inflammation and osteoclasia. Research3The Modulation Effect of BS on Treg and Th17cells of CIA rat modelAim:To investigate the effect of BS on Treg cell, Treg cell related cytokines and Th17cell related cytokines of CIA rat modelMethod:42female SD rats were randomly assigned to six groups, C group, M group, BSL group, BSM group, BSH group and TG group. Arthritis would be induced on day1. From Day13, the rat was administrated with BSL120mg/d/kg, BSM240mg/d/kg, BSH480mg/d/kg, TG24mg/d/kg and saline for18days. At day31, rats were sacrificed to perform (1) Flow cytometry to analyze peripheral percentage of Treg cell;(2) RT-PCR to analyze RNA quantity of Foxp-3, TGF-0, IL-6and IL-17RNA;(3) ELISA to detect serum IL-6and IL-17.Results:(1) BS could up regulate presentation of CD4+CD25+Treg cells and CD4+CD25+Foxp3+activated Treg cells in peripheral circulation;(2)BS could up regulate Foxp3, TGF-βRNA presentation and down regulate IL-6, IL-17RNA presentation;(3) Serum levels of IL-6and IL-17presented downward trend while the dose of BS was raised.Conclusion:The up regulation of Treg cell presentation would result in suppression of Th17cell which would lower the production of inflammatory cytokines to relieve joint inflammation. Research4The Modulation Effect of BS on Wnt Signaling pathway of CIA rat modelAim:To investigate the effect of BS on Wnt Signaling pathway of CIA rat model Method:42female SD rats were randomly assigned to six groups, C group, M group, BSL group, BSM group, BSH group and TG group. Arthritis would be induced on day1. From Day13, the rat was administrated with BSL120mg/d/kg, BSM240mg/d/kg, BSH480mg/d/kg, TG24mg/d/kg and saline for18days. At day31, rats were sacrificed and serum was obtained for ELISA to detect serum DKK1.Results:Serum levels of DKK1presented downward trend while the dose of BS was raised. The level is significantly lower than M group (p<0.01). Conclusion:BS could have modulation effect on Wnt signaling pathway to relieve joint inflammation and osteoclasia. Research5The Modulation Effect of BS on osteoclasia of CIA rat modelAim:To investigate the effect of BS on osteoclasia of CIA rat model Method:42female SD rats were randomly assigned to six groups, C group, M group, BSL group, BSM group, BSH group and TG group. Arthritis would be induced on day1. From Day13, the rat was administrated with BSL120mg/d/kg, BSM240mg/d/kg, BSH480mg/d/kg, TG24mg/d/kg and saline for18days. At day30, X-ray was performed. At day31, rats were sacrificed to perform (1)HE stain for ankle joint;(2) ELISA to detect serum IL-1, type II Collagen antibodies, cartilage oligomeric matrix protein, matrix metalloproteinase-3, C-telopeptide of type I collagen、Cathepsin K, propeptide of type-I procollagen and osteocalcin. Results:(1) BS could relieve radiological score of soft tissue edema and osteoclasia(p<0.05);(2) BS could relieve HE score of cellular infiltration, synovial hyperplasia and osteoclasia(p<0.05);(3)BS could down regulate serum level of pro-inflammatory cytokines(IL-1, TNF-a), type I collage degradation markers(MMP-3, ICTP, Cath-K), type Ⅱ collagen degradation markers(anti-Coll II, COMP)and bone remodeling markers(OC, PINP).Conclusion:BS could relieve joint inflammation, osteoclasia, suppress degradation of type I and type II collagen and modulate bone remodeling to prevent the occur of osteoclasia. Chapter4ConclusionBased on clinical observation and study, RA patients often present with deficiency of kidney. Therefore, Prof. Hou Liping formulated Bu Shen Tong Du capsule based on personal experience and experience of Qing famous Chinese Medicine practitioner Shi Guangji. Bu Shen Tong Du capsule is constituted of Yang He Tang and Semen Strychni which are effective in treating RA. The major compositions of the formula are Rehmannia Glutinosa, Colla Cornus Cervi, Cinnamomum Cassia Presl, Semen Sinapis, Semen Strychni and Ephedra Sinica Stapf. The formula applied pathogens dispersing herbs with enhancing and nourishing herbs to expel pathogens and enhance deficiency at the same time. The experiment showed that BS could relieve collagen-induced arthritis through modulation of immunity and bone and cartilage metabolism. The results elaborated and explained the therapeutic mechanism of BS. Since clinical study indicated that the combination of TG and Yang He Tang could effectively relieve clinical symptoms, we could also consider the combination of TG and BS in the future treatment of RA. |
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