The Primary Study Of Antisense Upar/pcdna3.1 (-) Expressing Plasmid To The Synovial Extracellular Matrix Of Collagen-induced Arthritis In Rats

Object:To investigate the effect of the uPAR Antisense RNA expressing plasmid to the synovial extracellular matrix of collagen-induced arthritis in ratsMethods:1. Extraction plasmid: Using the traditional alkaline method to isolate plasmid DNA from the bacterial solutions.2. 50 female Wistar rats were divided into 5groups equally: normal control group, model control group physiological saline treatment group,vector plasmid treatment group and recombinant plasmid treatment group .The latter 4 groups was injected intradermally chicken type-Ⅱcollagen to establish the CIA modle .After the CIA model was made successfully, physiological saline treatment group by injecting physiological saline, vector plasmid treatment group by injecting pcDNA3.1(-)plasmid and recombinant plasmid treatment group by injecting antisense uPAR/pcDNA3.1(-) expression plasmid.3. Observation of result:All the rats were selected to observe the general condition,arthritis index,pathological changes by H-E stained and the expression of uPAR and MMP-3(Matrix Metalloproteina-3) detected by immunohistochemical method.Results:1 Agarose gel electrophoresis showed that the recycling plasmids and the DNA marker were in the rather right position. The ratio of A260/A280 was 1.9-2.0,and the total plasmids was considered to be in good purity.The concentration of the plasmids was 510ng/ul,500 ng/ul.2 With the time lasting,the arthritis index of model control group,physiological saline treatment group and vector plasmid treatment group rose gradually and recombinant plasmid treatment group rose slowly; Comparing with the normal control group, there were synovium thickened lightly,synovium new vascnlar started to proliferate and inflammatory cells in synoviuar tissues in model controll group。3 The expression of uPAR:there was no expression of uPAR in nomal group.There was no significant difference between model control group,physiological saline treatment group and vector plasmid treatment group(P>0.05) and was significant difference between recombinant plasmid treatment group and other CIA group(sP<0.01);The expression of MMP-3:there was no expression of MMP-3 in nomal group.There was no significant difference between model control group, physiological treatment group and vector plasmid treatment group(P>0.05) and was significant difference between recombinant plasmid treatment group and other CIA groups(P<0.01)。The uPAR,MMP-3 were in positive correlation(r=0.604,P<0.01)。Conclusion:1. The antisense uPAR/pcDNA3.1(-)expressing plasmid could inhibit the expression of uPAR.2. The antisense uPAR/pcDNA3.1(-)expressing plasmid could inhibit the expression of MMP-3 and the degradation of ECM, and could provide an theoretic strategy for RA patient clinically.