The Clinical Observation And Experimental Research Of Knee Osteoarthritis Treated By The BuShenHuoXue Formula

ObjectivesOsteoarthritis of the knee (KOA), also named as hyperplasive gonarthritis, or osteoarthrosis of the knee, is a common degenerative articular disease, characterized with pain and stiffness of the knee, and fractional sound when the knee is moving. KOA is a disease of bone, synovium and the articular supporting structure. The incidence of KOA is upgrading yearly with the aging of world population and the incidence is also high in China.The pathogenesy of KOA is still haven' t been totally explained though some deep research had been done on it. It is also the reason why there is no ideal treatment cause all the cure method is symptomatic treatment. Chinese traditional medicine (TCM) has the better effect of treating early stage of OA and it has the dominace of cheaper, less by-effects and simultaneous treatment of principal and subordinate symptoms. My advisor professor Deng Jinfeng created the principle of invigorating the kidney and promoting blood flow. He drew up the BuShenHuoXue Formula based on the renal deficiency and blood stasis pathomechanism of KOA. The formula has satisfied clinical therapeutic effect in clinical long-term use. The principle of evidence-based medicine was used in this research and the research was designed with randomization and comparation. A systematic observation of clinical curative effect of KOA treated by the BuShenHuoXue Formula had been done in the research, with the study of the effect of drug contained serum of the BuShenHuoXue Formula(DCS-B) on chondrocytes and synoviocytes cultured in vitro was made to explain the curative mechanism of the Formula at the molecular level.Contains 1. Clinical Research1.1 General Data 68 patients with early stage of KOA was seperated into 2 groups by the principle of randomization and comparation. One group is treat group of the BuShenHuoXue Formula; the other group is controlled group of celecoxib.1. 2 Cure Method Patients take according medicine for 4 weeks as one course of treatment, curative effect been analyzed after 2 courses. Index of severity of osteoarthritis (ISOA) and serumal TNF-a and IL-1P level was observed.1.3 Results The patients of treatment group and control group are comparable in age, gender and course of disease, according to the statistical treatment(P>0. 05). Clinical therapeutic effect analyze shows both groups have the effect of obviously relieving pain, improving the knee joint function. The total effective rate of bushenhuoxue formula is 88.89% and celecoxib is 90.62%. The ISOA and serumal TNF-a and IL-13 level of both group before the treatments has significiant defference(P<0.05) when compared with the results after the treatments, while there is no significiant difference between groups (P>0.05). No poison and side effect was found during the treatments.2. Experimental Research2.1 Manufacture of drug contained serum of the BuShenHuoXue Formula (DCS-B) Intragastric administration for 7 days based on the conversion of drug dosage between human and rice, then the blood obtained, its serum separated and reserved. Drug contained serum of Celecoxib was made at the same time.2.2 Separation and culture of primary chondrocyte and synoviocyteMechanical and enzymatic digestive combined method was used to separate and harvest primary cartilage and synovial cells and adherence regulation used to purificate synoviocyte. Cartilage and synovial cell line were successfullybuilt with the adherence culture method.2.3 The effect of DCS-B to the proliferation of chondrocyteCell groups was devided according to the different density of DSC-B, then the chondrocyte get synchronized and MTT was performed to detect the activity and proliferative ability of the chondrocyte. Growth curve of chondrocyte was drawn. The difference level of different density of DSC-B was analyzed based on the 6d' s OD value measured by the enzyme scale meter.2.4 The effect of DCS-B to the TNF-aand IL-1 P level of synoviocyteThe synovoicyte within third generation was used to do the research. LPS was used to induce the generation of TNF-a and IL-13, then DCS-B and drug contained serum of Celecoxib put into intervention. Comparation of the TNF-a and IL-1P level was made among different density of DCS-B and between TCM and west medicine.Results1. The curative effect of the Bushenhuoxue Formula and Celecoxib was compared through the random and controlled experiment of 68 patients with early stage of KOA. The result shows both the BuShenHuoXue Formula and Celecoxib have the obvious effect of reliving pain and improving knee joint function with the total effective rate of the Formula is 88.89% and Celecoxib is 90.62%. There is no significant difference between the two groups due to a Ridit analysis. Significant difference of ISOA and serumal TNF-a and IL-1 P level was found when compared with pretherapy in both groups (P<0.05) while no significant difference between the two groups (P>0.05).2. Seperation and culture of chondrocyte and synoviocyte in vitro was successfully done and cartilage synovial cell line established. Toluidine blue stainning used on cartilage cell and Giemsa stainning used on synovial cell to identify its phenotype led to satisfied results. The affect of passage to cell phenotype was studied with the conclusion that cells within 3 generations are recommended for experimental research.3. With the method of serum pharmacology, MTT way was used to detect the activity and proliferative ability of chondrocyte for the study of the effect of the BuShenHuoXue Formula on the proliferation of cartilage cell. The result...