The Effection Of KJA To Rat IL-6 Gene At MRNA Level And Its Construction Of SiRNA Expression Vector

Objective:To investigate the expression of intedeukin 6(IL-6)in intestinal tissue of rat with Ulcerative Colitis(UC)and further studies the pathogenesis of UC.To construct the specific small interfereing RNA(siRNA)expression vector that can block the rat IL-6 gene by RNA interference(RNAi)technique,which provides a new approach to determine the onset gene of Ulcerative Colitis and investigate the target of the Uygur medicine Kuijie'an(KJA)that can treat the UC.Methods:Wistar rats were derided into 5 groups:control group,UC groups,NS group,5-ASA group,UC rats induced by DNCB and acetic acid.Observe and assay the expression of IL-6 gene by RT-PCR.According to the encoding sequence of mRNA of rat IL-6 provided by Genbank,two pieces of specific hairpin structure short chain oligonucleotide sequences were designed and synthesized by using the siRNA software.The annealed oligonucleotide fragment were subcloned into pSilencer ^TM1.0-U6 expression vector by gene clone technique and transformed into DH5αE.coli.Finally the plasmid evaluated by using enzyme cutting and sequencing. Results:Expression of IL-6 gene remarkably increased in intastinal tissue of rats with UC as compared with control group;but it remarkably decreased after theraphy of KJA. Identification using enzyme cutting and sequencing showed that the IL-6 siRNA expression vector was constructed successfully.Conclusion:It demonstrated tentavtively that KJA had an effect on UC.Successful construction of rat IL 6 siRNA expression vector will facilitate further studies of determine the onset gene of Ulcerative Colitis and investigate the target of the Uygur medicine KJA that can treat.the Ulcerative Colitis.