Study On The Effect And Mechanism Of Tefimide A On Rheumatoid Arthritis

Rheumatoid arthritis is a chronic systemic inflammatory autoimmune diseases,which is characterized by joints and joint tissue inflammation main performance for joint synovitis.Rheumatoid arthritis(RA)disease is a continuous,repeated process which patients are suffering from.The main treatment for RA consists of medicine therapy and surgical therapy.At present,western medicine therapy include nonsteroidal anti-inflammatory drugs(nonsteroidal anti-inflammatory drugs,NSIADS),rheumatism drug(diseases-modifiying antirheumatic drugs,DMADs),glucocorticoid(glucocorticoids)and biological preparations(TNF-α inhibitor,IL-1β blockers),etc.Although NSIADS and glucocorticoids can effectively reduce joint inflammation,but do not prevent bone destruction process.DMADs is limited for its long cycle and side effects.Biological agents considered to have pretty effects are effective for 70%RA patients and expensive.So seeking for compensatory and alternatve drugs is becoming more and more important for the treatment for RA.Chinese materia medica monomers have many advantages,such as little side effects and low price,becoming the impotant source of RA alternative drugs.Tubeimoside-1(TBMS I),a natural compound isolated from Bolbostemma paniculatum,which has been widely used as a traditional Chinese herbal medicine,has anti-tumor effects.Besides,Jiake Xu,laboratory found that TBMS I significantly inhibited the osteoclastogenesis and resorption activity of osteoclast.Osteoclast is the main damage of RA bone damage cells.In view of this,we supposed that TBMS I might be involved in the pathogenesis of RA process and has a certain inhibitory effect.To this end,we analyzed through experiments in vivo and in vitro.TBMS I possible therapeutic effect of RA,and further discussed mechanism.Therefore,we proposed that if TBMS I had a moderating effect on arthritis and investigate the mechanism on the level of cellule,cell,and animal.To this end,we established a type II collagen-induced arthritis(CIA)Rat model,analyzed the incidence of the experimental group,confirmed that TBMS I had the regulatory effect on RA.The main contents and results of the experiment:PartⅠ:In vitro experimentsCultured synovial fibroblsts(fibroblast-like synovial cells,FLS).The toxicity of TBMS I was tested by MTS/PMS assay;The infuences on cell migration and proliferation were tested by cell wound assay and EdU test;The intracellular cytokine in FLS stimulated by TNF-α were detected by RT-PCT.The influence of TBMS I on the MAPK signal pathway was detected by Western blot.Result:TBMS I(OμM、0.5μM、1.0μM、2.0μM、5.0μM)had no inhibition on cell survival;TBMS I had inhibition on the migration of FLS stimulated by TNF-α when their concerntration were more than 0.5μM;TBMS I could down-regulate inflammatory factors,MMP-9;TBMS I had an inhibition on the proliferation of FLS stimulated by TNF-α when the concerntration were 1μM.results showed that cytokines were inhibited by the inhibition of JNK and p38 phosphorylation.Part Ⅱ:In vito experimentsSelected 160g-180g female Wistar rats to induce arthritis model to build rheumatoid arthritis modle by CIA methods.Experiment was divided into five groups:① Normal group(Control):do not induce RA,intraperitoneal injection of saline solution;②CIA control group(CIA+Vehicle):induction of RA,intraperitoneal injection of saline solution;③ low-dose drug experiment(CIA+TBMS I 1mg/kg):induction of RA,intraperitoneal injection of TBMS I(1mg/kg);④ middle-dose drug experiment(CIA+TBMS I 5mg/kg):induction of RA,intraperitoneal injection of TBMS I(5mg/kg);⑤high-dose drug experiment(CIA+TBMS I 10mg/kg):induction of RA,intraperitoneal injection of TBMS I(10mg/kg).Early onset(hind paw score of 1-2 points)began drug treatment,intraperitoneal injection of saline,different concerntration of TBMS I,once a day.The total treatment time was 14 days.After treatment,rats were sacrified by euthanasia.The differences between the experimental and control groups in terms of the degree of swelling,the expression of inflammatory cytokines,joint destruction were compared to identify different dose of TBMS I effects on rheumatoid arthritis.Results:Low dose of TBMS I could not effectively ease the severity of CIA Rat.But middle and high dose of TBMS I were able to alleviate joint swelling as well as the concerntration of TNF-α and IL-β in CIA rat serum in a dose dependent manner.Pathology slice and X-ray results showed that 5mg/kg TBMS I and 10mg/kg TBMSI could ease joint synovial inflammation,and inhibit the formation of the synovial tissue hyperplasia and pannus,as well as cartilage and bone tissue damage in a dose dependent manner.Conclusion:Cell level and the level of animal experiment showed that the TBMS I has therapeutic effect on rheumatoid arthritis by reducing inflammation,restraining synovial cell proliferation and migration,as well as the bone tissue damage.