| Objective: Establishment of expelling wind, removing heat and promoting bloodcirculation to remove meridian obstruction method. Preparation containing serumcontaining JBY, and use serum containing to stimulate synovial cells, using serumcontaining synovial cells stimulated by MTT assay, flow cytometry and other methodsto detect the drug-containing serum on synovial cells which play a vital role with acolorimetric detection of different concentrations of JBY the role of the drug in theserum-containing synovial cell activity impact on Caspase-3.Arthralgia further explanation JBY treatment efficacy, as JBY application provides areliable experimental basis in clinical practice.Methods:(1) Establish JBY treatment Arthralgia therapies, theoretical discussionexpelling wind and removing heat and promoting blood circulation to removemeridian obstruction method.(2)30breeding female SD rats used to extract thedesired serum containing drug.(3) The30female SD rats were randomly divided intothree groups, each fed with different concentrations of JBY and controls as Saline. SDrats were observed activity,diet,sleep,etc. In general, the eye after taking blood, getJBY containing serum.(4) And it will be our hospital joint replacement patients withRA synovium and synovial tissue of OA patients were cell culture experiments using3-4synovial cells with different concentrations of serum containing synovial cellsstimulated simultaneously with normal saline control group.(5) Impact on thedetection of synovial cell proliferation, flow cytometry, detection of synovial cellcycle and apoptosis by MTT assay. Confirmed JBY containing serum can promoteapoptosis of synovial cells in rheumatoid arthritis, and osteoarthritis synovial cellsineffectively.(6) With a colorimetric assay JBY serum containing differentconcentrations of the drug in the role of synovial cells (Caspase-3activity), and toexplore aspects of the induction mechanism and the role of apoptosis.Results:(1)MTT results showed that: different concentrations of JBY on RA synovialfibroblast proliferation was inhibited, especially in the high-dose group wassignificantly(P<0.01). RA inhibition of each group over time,and gradually increasingthe dose increased,showing the time and patience by the dose(P<0.05). OA synovialcell growth inhibition at low dose is not obvious, but as the dose is increased and prolonged effect on synovial cells lighter inhibition(P <0.05).(2) Flow cytometryshowed: JBY drug containing serum concentration of20μl/mL,30μl/mL,40μl/mL,when50μl/mL could induce apoptosis in synovial fibroblasts (P<0.01). Comparedwith the control group, apoptotic cells JBY containing serum group was significantlyincreased, and there was a significant difference(P <0.01) increased.JBY induced byRA synovial cells is superior to induce apoptosis in synovial cells.(3) JBY make cellcycle arrest in G0/G1phase and has a concentration and time dependent.(4) AssayShow: With JBY with increased serum concentrations, Caspase-3activity in synovialcells of each group were gradually increased,the concentration of the effective rolefrom20umol/L began to stimulate activity increased the longer, the more obvious.This results suggest that Caspase-3may be involved in the process, JBY synovialfibroblasts induced apoptosis,the process may be by way of Caspase-3.Conclusion:JBY can inhibit the growth of synovial cell proliferation, in a certainrange of time, dose showed some correlation. JBY make cell cycle arrest in G0/G1phase and has a concentration and time dependent. JBY can induce apoptosis ofsynovial cells, and its mechanism may be associated with activation of Caspase-3pathway. The RA clinical trials and drug screening reference dose, in order to promotebetter service for the majority of patients. |
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