Study On Ikkγ Gene Changes Of Marc-145 Cells Infected By PRRSV

Objective: To study whether the IKKy participate in the immune reaction mechanism of anti-PRRSV process, we established the model of Marc-145 Cells infected with PRRSV and detected the variations of IKKy gene expression.Methods: We used the Indirect immunofluorescent assay (IFA) to evaluate whether the model has been successfully established, detected LY294002 inhibition rate towards the model cells using MTT method. The model cells were divided into 4 groups: Cells control group (treat with nothing), LY294002 group (treat with LY294002), PRRSV group (treat with PRRSV) and virus+LY294002 group (treat with PRRSV and LY294002). We also detected IKKy gene and protein changes of Marc-145 cells infected with PRRSV when treated with or without LY294002 using qPCR and Western blot, thus we studied that the molecular immune mechanism of IKKy participate in anti-PRRSV process.Results: ①We used the Indirect immunofluorescent assay (IFA) to make sure the model has been successfully established.②MTT method results proved that compared with cell control, PI3K inhibitor LY294002 can inhibit the growth of Marc-145 cells, the inhibition rate and the concentration of the inhibitor are positively correlated. The inhibition rate raised in 48h, however, after 48h, the inhibition rate has decreased. ③The results of the qPCR showed that, compared with the control group, the IKKy mRNA expression of the PRRSV group has raised (P<0.05), the IKKy mRNA expression of the LY294002 group has decreased (P<0.01), the IKKy mRNA expression of the LY294002+PRRSV group has decreased (P <0.01).④The results of Western blot shows that the IKKy protein expression of the PRRSV group has raised, the IKKy protein expression of the LY294002 group has decreased,the IKKγ protein expression of the LY294002+PRRSV group has decreased.Conclusion: ①The upstream signaling pathway affected the signal transduction of the downstream signaling pathway.② LY294002 inhibitor affected the transcription and replication of PRRSV. ③IKKγ took part in the reaction of host anti-PRRSV.