Observation On Therapeutic Effects Of Tongbi 2 Mixture And Research Its Contribution On B7/CD28

Objective:Rheumatoid Arthritis(RA)is a disease characteristic with uncertain etiopathogenisis, chronic,symmetric and polyarthritis,which could involve in other organ and tissue,such as skin,muscle,blood vessel,nerve,membrana pleuralis,cardiac muscle and lymphoid node. RA invades and destroys the joints,which is similar to tumor,induces various arthrocleisis, anisotrophy and functional incapacitation ultimately during the development of disease.RA is a common disease with high mutilation rate.It has severe influence on quality of life. The doctors seek its source of pathopoiesis and effective therapeutic regimen actively in order to lessen labor force loss and mutilation,radical cure the RA and benefit for mankind. There is not such a disease named RA in traditional Chinese medicine.RA can be category into "Bi Bing","Lijie Bing","Fengshi","Hexifeng","Baihu Lijie" according to its clinical manifestations.It's considered that Bi Bing was caused by wind,cold and dampness invaded into body together by the theory of traditional Chinese medicine.The Golden Cabinet created the decoction of Guizhi Shaoyao Zhimu and the decoction of Wutou to cure Lijie which sufficiently reflected dominance of treat the RA with Chinese medicine.Tongbi 2 mixture is composed by Nansheteng and Chuanxuduan which promote therapeutic effect mutually.It can warm meridians and disperse coldness,expel wind-evil and remove wetness,tonify liver and kidney,activating meridians to ease pain.As a means to treat RA, the mixture is not only belongs to China but also to the world.Thus,it's necessary to discuss the mechanism on treat RA of Tongbi 2 mixture.T lymphocyte activation,synovial cell multiplication,inflammatory cell infiltrate, inflammatory factor anomal secretion causes synovial multiplication and forms pannus in the pathogenesy of RA.A series of clinical manifestations,such as arthrocele,arthrodynia and arthrodial cartilage damage,occur and even multisystem is involved in.Autoantibody and some inflammatory eytokine are correlated with occurrence and development of RA. B7/CD28 molecule becomes the key signal in occurrence and development of autoimmune disease since it participate in the composition and expressing regulation of autoantibody and inflammatory cytokine with its costimulation.It prognosticates B7/CD28 might be ideal target in treating autoimmune disease.The topic,which reveals effective target and mechanism of traditional Chinese medicine from the view of molecular biology,systemically analyzes mechanism of Tongbi 2 mixture on treating RA via studying its influence on co-stimulating molecule,cytokine,synovial cell,arthrodial cartilage damage and clinical effect.It offers reliable data for developing new medicine to treating RA and applying the mixture systemically and normally.Methods:Part of experimental studyingForty specific-pathogen free rats were divided randomly into normal control group,Tongbi 2 mixture-treated group,MTX-treated group,TWP-treated group and untreated group,8 rats in every group.Except the normal control group(injection with normal saline at the same dose),the other groups were raised collogen induced arthritis(CIA)model through subcutaneous injection with collagen proteinⅡ.The groups were treated respectively with normal saline,Tongbi 2 mixture,MTX tablets and TWP tablets for 36 days.1.All of rats get blood from abdominal aorta.The expression of CD28 and CD152 was detected by flow cytometer,2.All of rats get 2ml blood from abdominal aorta and separate serum.Every sample set up 2 ambi-hole.Tumor necrosis factor-αwas analyzed by enzyme linked immunosorbent assay.3.Hind limb was token X-ray photo with 60kV voltage,0.04ms expose time,7mA current. Checked and analyzed imaging with score of radiology standardization recommended by American College of Rheumatology(ACR).4.Synovium was retained by 4%polyoxymethylene,imbedded by paraffin,4um slice, stained with hematoxylin and eosin and detected pathology with optical microscope.Part of clinical research1.Diagnostic criteria.Reference the separate criteria of ACR 1987.2.Experimental medication.Tongbi 2 mixture was decocted at 0.625g dried medicinal herb per mini-liter by manufacturing laboratory of first affiliated hospital,Guangzhou University of TCM.45g Nansheteng was decocted in advance for 3.5h while with Chuanxuduan for 0.5h more.Take medicine twice a day.3.All 60 selected cases,which was separated 40 cases for treat group and 20 cases for control group,were the patients in the department of rheumatism and immunization,first affiliated hospital,Guangzhou University from Apr,2004 to Jan,2008.The treat group was treated with Tongbi 2 mixture 100ml twice a day while control group was treated with MTX 10mg once a week correspondingly for 30 days.Recorded observed index.4.Observed index of clinical effect.(1)Biological parameter include in gender,age,height and weight.(2)Safe parameter contains blood,urine routine,hepatic function and renal function.(3)Effective index.Cardinal symptoms and physical sign:number of tender and swelling joints,morning stiffness,average grasp force of both hand,walking time for 20 m, health assessment questionnaire(HAQ)and visual analogue scale(VAS).Laboratory test: rheumatoid factor(RF),erythrocyte sedimentation rate(ESR),C-reactive protein(CRP).ResultsThe expression of CD28 among drug treated groups is same(P＞0.05),while higher than normal control group(P＜0.01)and lower than untreated group(P＜0.01).The expression of CD152 on Tongbi 2 mixture treated group is lower than MTX and TWP treated group (P＜0.050),but is same to normal control group(P＞0.05).It's same on expression of TNF-αbetween drug treated and normal control groups(P＞0.05),lower than untreated group coincidently.Synovium of joint,cartilage pathology and imaging of joint among drug treated groups is same(P＞0.05),while different from normal control group and untreated group(P＜0.008).Total effective rate of treated group is 80%,meanwhile 70%of control group.It's significant difference with pretherapy in number of tender and swelling joints,moming stiffness,average grasp force of both hand,walking time for 20 m,HAQ and VAS (P＜0.001 or P＜0.05).Compared with control group,physical signs are same except the number of tender joints.ESR,CRP and RF are obviously lower than that of pretherapy of treated group(P＜0.001 or P＜0.05).ESR and CRP are obviously lower than that of pretherapy of control group(P＜0.001)except RF has no significant difference(P＞0.05).It has no significant difference in improving ESR,CRP and RF compared with control group (p＞0.05).Conclusion1.It cues that Tongbi 2 mixture has favorable therapeutic effect on RA since it can improve the number of tender and swelling joints,morning stiffness,average grasp force of both hand,walking time for 20 m,HAQ and VAS of RA patients.2.Tongbi 2 mixture has satisfied therapeutic effect on treating inflammatory of synovial tissue of joint.It can inhibit activation and differentiation of T lymphocytes through regulating expression of CD28/CD152,and regulate expression of TNF-αdown to physiological level,thus decrease expression of other inflammatory cytokine.It can reduce production of pannus and arthromeningitis,lessen osteolysis,promote tissue repair.That may be the major mechanisms in curing RA.3.It's considered that the pathogenesis of RA is wind,cold and dampness which invading the body to cause phlegm and stagnant blood hinder into tendon and bones mutually,results in tendon injury and bone damage ultimately.Tendon injury and bone damage is tendency of RA.Thus,therapeutic method of RA is to warm meridians and disperse coldness,expel wind-evil and remove wetness,tonify liver and kidney,activating meridians to ease pain.4.Based on the theory of traditional Chinese medicine,it's regarded the compatibility feature of Tongbi 2 mixture is expelling wind and remove wetness,deoppilation meridians to ease the arthralgia spasm;tonify liver and kidney to prevent tendon injury and bone damage;refined compatibility and great clinical effect.