Experimental Study On The Gene Expression Profiles Of Triptolide Affect AD Joint Tissue Of Rats

Objective To study the molecular biological foundation for the mechanism on the inhibition of newborn RA synovial pannus by triptolide,We detect and verify differential expressed genes from synovial tissues of AA model group and triptolide treatment group by gene transcripts produced by Agilent.Methods 20 male Wistar rats were randomly assigned into model group and triptolide treatment group,10 in each group.To create AA model,all the rats were injected with 0.1ml,FCA(ip)everyday for 18 days,and scored by arthritis index assessment.After the AA model was achieved,rats of 2 groups were injected with triptolide 0.4mg/kg,and equivalent amount of saline respectively for 14 days.After the treatment,synovial tissues in ankle joint of rat’s hind legs were obtained for gene express detection.Testing requirements for processing as follows:Total RNA was extracted and purified by reverse transcription synthesis of a fluorescent molecule(Cy3/Cy5)cDNA probes labeled using rat genome Agilent microarray hybridization with the hybridization signal by scanning and digitizing presumed triptolide on gene expression profiles of rheumatoid arthritis and possible signaling pathways through bioinformatic data analysis.And treated with RT-PCR detection of synovial tissue Vegfa,Clqtnf3 mRNA expression of.Results data SPSS 17.0 software for processing and statistical analysis.Conclusion(1)According to AI(arthritis index)scoring assessment,the difference between 2 groups is unsignificant with P>0.05 in 18th day,while significant in 32th day with P<0.05.After treated with triptolide,there is a positive effect on rats of treatment group with P<0.05.(2)Press differential gene screening criteria,select Ratio value>2 is up-regulated genes,Ratio value<0.5 to down-regulate genes.There were 48 genes differently expressed after selected by gene screening and 32 genes up-regulated,while 16 genes.down-regulated.(3)RT-PCR detection reveals that genes Vegfa,Clqtnf3 down-regulated.in treatment guoup compared with model group with P<0.05.Results Firstly,Treatment of triptolide has a influence on gene expression of AA model rats.After detection.Compared with model group,there were 48 genes differently expressed after selected by gene screening and 32 genes up-regulated,while 16 genes.down-regulated.in triptolide treatment guoup.PI3K-AKT signaling pathway plays a significant effect on proliferation and apoptosis of synovial cell in RA joint after analized by GO and Pathway.Secondly,After the expression level of 2 genes are detected by RT-PCR,and the result proved to be consistant with gene chip detetion,it means Our data is reliable.Thirdly,triptolide could play an inhibition role in the expression of genes Vegfa and Clqtnf3,which are closely related to occurrence and development of RA.