Study On The Influence Of Nrdp1 And SOCS-1 For Brucella Intracellular Survival And Macrophage Apoptosis Induced Bybrucella Infecting

Brucella is kind of pathogenic bacterium that can cause zoonotic disease. The main goal of brucella infection is professional and non-professional macrophages,to survival and reproduction in the cells.The molecular mechanism of interaction between pathogen and host cells is complicated, also the cause of brucellosis.So intracellular survival of brucella and molecular interactions mechanism research between the host and the pathogenic bacterium is significance. The ubiquitin-proteasome pathway is involved in the regulation of variety intracellular physiological and biochemical reactions.This study concentrate on the influence of E3 ubiquitin ligase Nrdp1 and SOCS-1 among brucella infection.Objective:The study based on genes of mice macrophage Nrdp1 and SOCS-1,to interfere and overexpressed the genes,and then brucella melitensis 16 M infects the cells,to explore their influence to intracellular survival of brucella and host cell apoptosis caused by brucella.Methods:(1)Brucella melitensis 16 M infect mice macrophage RAW264.7 and then q RT-PCR to detective the expression of Nrdp1 and SOCS-1 after 0h、2h、4h、8h、12h and 24 h by the infection.(2)Interfering and overexpressing the expression of Nrdp1 and SOCS-1 in mice macrophage RAW264.7,and then brucella melitensis 16 M infect the cells to detective CFU, percentage of apoptotic cells and expression of genes Bax,Bcl-2,TNF-α.(3)Interfering and overexpressing the expression of Nrdp1 and SOCS-1 in mice macrophage RAW264.7,and then detective the expression of genes JAK1、JAK2、JAK3、STAT1、STAT2 and STAT3.Results:(1)The expression of Nrdp1 and SOCS-1 shows a regular change as time goes on.As the cells infected by brucella melitensis,expression of the genes were up-regulated rapidly,then were down-regulated effected after 4h.Up-regulated at 4-8h,down-regulated at 8-24h(P<0.05).(2)Pick one better from each two interfering models,which percentage of interfering is 66% for PLL3.7-N1 and 54% for PLL3.7-S1.And the percentage of overexpressing is 228% for Plex-Nrdp1 and 332% for Plex-SOCS-1.CFU experiment shows that the log CFU of Plex-Nrdp1 and Plex-SOCS-1 are remarkable lower than control groups.But difference from PLL3.7-N1 or PLL3.7-S1 and the control groups is not remarkable.The expression of Bax and Bcl-2 for Plex-SOCS-1 is remarkable rised,but Bax/Bcl-2 is not.Bax is remarkable lower and Bcl-2 is not for PLL3.7-S1,and Bax/Bcl-2 is remarkable lower.Bax and Bcl-2 for Plex-Nrdp1 is remarkable rised,and the Bax/Bcl-2 too.Bax for PLL3.7-N1 is remarkable lower and Bcl-2 is not,and Bax/Bcl-2 is lower.TNF-α for Plex-SOCS-1 is rised,and PLL3.7-S1 is lower.And difference of Plex-Nrdp1 and PLL3.7-N1 is not remarkable.The percentage of apoptotic cells experiment shows that Plex-SOCS-1 is rised,and PLL3.7-S1 is lower,and Plex-Nrdp1 is rised and PLL3.7-N1 is not remarkable.(3)The expression of JAK1、JAK2、STAT1、STAT2、STAT3 for Plex-SOCS-1 is lower and JAK3 is no difference. JAK1、JAK2、STAT1、STAT2、STAT3 for PLL3.7-S1 is rised and JAK3 is no difference.JAK1、JAK2、STAT1、STAT2、STAT3 for Plex-Nrdp1 is lower and JAK3 is no difference.JAK1、JAK2、JAK3、STAT1、STAT2、STAT3 for PLL3.7-N1 is no difference.The results show that:In the infection of brucella,Nrdp1/SOCS-1 inhibit brucella to intracellular survival,and induce apoptosis of the host cells,at the same time, play a negative regulatory role to the signal path of JAK/STAT.To sum up,Nrdp1/SOCS-1 is important for brucella to intracellular survival and apoptosis induced by brucella among brucella infecting.