| Osteoarthritis(OA)is a chronic, retrogressive article disease that seriously does harm to patient quality of life.The incidence of Knee Osteoarthritis(KOA)is the highest in all joints of humanbody, and the pathological changes of arthrodial cartilage and surrounding soft tissues are very important in KOA. At present, Western medicine is lack of effectively therapeutic method.Besides,the effects of the traditional Chinese medicine on soft tissue is firmly, but the persuasively experimental study is still blank. So, by optical microscopy, electron microscopy,biochemistry method, immunohistochemistry method, and other advanced means,the empirical study uses combinative methods of TCM and Western medicine, systematically and comprehensively explore therapeutic effects of gubixiao on KOA. From the whole (the KOA rabbit),cell (Apoptosis), molecule (SOD, MDA, NO) to gene level(DNA),The study enhances the scientific research capability,evaluates therapeutic effects quantificationally,which gives the study scientific nature, comparability,reliability, and at the same time it also illuminates the mechanism of therapeutic effects of"gubixiao"on impaired periarticular soft tissues of knee(for example:synovium).Objective: To explore the therapeutical effects of gubixiao on KOA of the empirical rabbits.Methods: 60 Rabbits were randomly divided into five groups:control group,model group,celecoxib group,gubixiao group,zhuangguguanjiewan group,Each group has twelve Rabbits. They are raised in the same environment,one week later,except control group Other four groups of rabbits are made animal model of KOA by extended fixation technique.medication:From the first day of being fixed,all rabbits are fed the same food and different medicines respectively. control group and model group: normal saline 2ml/Bid. p.o.;celecoxib group, celecoxib liquor 2ml/Bid.p.o.(55mg/d);gubixiao group, gubixiaoliquor 2ml/Bid.p.o.(688mg/d); zhuangguguanjiewan group, zhuangguguanjiewan liquor 2ml/Bid.p.o.(1650mg/d ).Examination indexes:â‘ Plasm SOD,MDA and NO of all groups of the rabbits were dividedly measured after the rabbits being treated for 6 weeks and 10 weeks by biochemistry method, and analyzed by spss 11.0 statistic software.â‘¡After being treated for 6 weeks and 10 weeks,the samples are took from arthrodial cartilage of tibia tuberosity and center synovium of knee .At first the samples are observed by naked eye,then they are fixed,dehydrated,embeded,sliced, stained for electron microscopy and optical microscopy observation.â‘¢At the same time,samples from center synovium of knee are homogenated .The clear supernatant liquid are taken to test SOD and MDA level, and then analyzed by spss 11.0 statistic software.â‘£After being treated for 6 weeks and 10 weeks arthrodial cartilage were taken randomly from five groups and ananlyzed apoptosis incidence (AI) by TUNEL. The expression rate of p53 and Bcl-2 of cartilage cell of all groups of rabbits wered analyzed by CIAS-1000 image analysis system software to explore the mechanism of action.Results:KOA animal model were made the by extended fixation technique,which is received worldwide. Models are proved to be successed. Biochemical index demonstrates that Plasm SOD,MDA,NO and synovium SOD and MDA of the rabbits change afer being treated for 6 weeks and 10 weeks. Plasm SOD level and synovium SOD level in Model group are lower than that of other groups,but plasm MDA,NO level and synovium MDA level are completely the opposite. Post-treatment the Plasm and synovium SOD activity increase,MDA and NO level decrease differently in degree,and group comparison has statistical significance.Macroscopic observation indicates that after being treated for 6 weeks,the model rabbits ,articular cartilage lose the original gloss,turn yellow,have few anabrosis dots,and the synovium tissues proliferate.After being treated for 10 weeks,the articular cartilages obviously lose the original gloss,turn yellow and gloom,the cartilage surfaces are uneven,periphery synovium obviously proliferate.But pathological changes of all treating groups are better than that of model group,among which the gubixiao group is the best.Electron microscopy and optical microscopy observation indicates that after being treated for 6 weeks ,cartilage cell of model rabbits pyknosis,cell is similar to cone,nucleus is highly pyknosised ,cellule is seen surround nucleus,cell organs disappear,cell membrane damaged,and patches are found in matrix under electron microscope.Apoptotic bodies are seen in the samples of model group after being treated for 10 weeks.Under optical microscope,cartilage cell disorder,cluster, swell,karyopyknosis and even dissolved,cellular content decrease,the collagen fibers of matrix are lightly fibronolysis after being treated for 6 weeks,while cartilage cell cluster obviously,karyopyknosis,nucelus dissolve,matrix stain unevenly or lightly,the collagen fibers are obvious after being treated for 6 weeks .Comparison of terms of the scores with cartilage pathological assessment have statistical significance. Under optical microscopy,the changes of synovium of model rabbits are obvious:epitheliums proliferate, phlegmasia cells infiltrate,vasculars invade below synovium after being treated for 6 weeks. After being treated for 10 weeks,the synovium epitheliums proliferate heavily,cells hypertrophy,large quantities of granulational tissues hyperplasia below synovium,a great deal of vasculars invade,a lot of phlegmasia cells infiltrate.Pathological changes of treating groups are lessener in different degree compared with model group,but gubixiao group is significantly lightener compared with that of celexibu group and zhuangguguanjiewan group.Expression of p53and Bcl-2: Under optical microscope,the cell image analytical system demonstrates that the expression of p53and Bcl-2 of cartilage cell of model group increased; Expression of Bcl-2 increases gradually in gubixiao,celexibu and zhuangguguanjiewan group following treating,while expression of p53 decreases.After being treated for 6 weeks and 10 weeks,the expression of p53 of gubixiao group descend compared with that of model group(P <0.01). While after being treated for 6 weeks and 10 weeks,the expression of Bcl-2 of gubixiao Group significantly increases. Group comparison between gubixiao Group and model group has statistical significance (P <0.01).TUNEL for apoptosis: Under optical microscope,The AI of the model group is significantly higher than that of other groups.Post-treatment the AI of the treating groups is lower in different degree.Gubixiao group is the lowest. Compared with model group the AI of gubixiao group is obviously lower (P <0.01) after being treated for 10 weeks.Conclusions:Gubixiao can clear surplus of free radicle, retrieve metabolic disorder of oxyradical;Gubixiao can facilitate metabolism of cartilage cell and help cartilage to recover which can relatively keep the integrity of cartilage structure; Gubixiao can inhibit phlegmasia changing of synovium, reduce phlegmasia liquid delivering into joint cavity,reduce destruction of phlegmasia synovium to cartilage;To some extent, gubixiao can block up path of inducing cartilage cell apoptosis, decrease apoptosis quantity of cartilage cells and help cartilage to recover. |
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