| Objective:1.The study is to investigate the signal pathway of Toll-like receptor signal(TLR-9)in B lymphocyte stimulating factor(BLyS)induced systemic lupus erythematosus(SLE)transgenic mice model,detect anti-dsDNA antibody titer,C3 and C4 complement levels,inflammation index IL-10 and disease activity ESR,CRP level.2.We detect the expression of TLR9 mRNA in SLE patients and the serum level of IL-10 and illuminate the association between these indexes and the activity of SLE.3.We also discuss the modulation of 1,25(OH)2D3 on SLE patients.All of the three parts are in order to provide basis for the pathogenesis and new therapeutic targets of SLE.Methods:1.The transgenic female mice of EB virus membrane antigen BLLFl were composed by 21 animals,with age of 8-10-week old and weighting 30-40g.The mice were randomly divided into blank control group,BLyS blocker group and TLR-9 blocker group,which formed by 7 animals each,while the blank control group was received intraperitoneal injection of equal amount of normal saline;Anti-BR3 monoclonal antibody 500μg/d/time was applied in BLyS blocker group,with continuous intraperitoneal injection for 10d;Anti-human TLR-9 antibody 250ng/d/time was adopted in TLR-9 blocker group,with continuous intraperitoneal injection for 10d.Peripheral venous blood should be taken after keep feeding for 10d.The relative expression of TLR-9 mRNA was tested by RT-PCR;BLyS protein concentration and IL-10 level were detected by ELISA.2.We collected 106 SLE patients to detect some clinical indexes.The relative expression of TLR-9 nRNA was tested by RT-PCR;BLyS protein concentration and IL-10 level were detected by ELISA.The immunization spots method is for detection of anti dsDNA,and conventional biochemical method is for detecting C3,C4 and ESR and CRP level.3.The study was carried out on patients with SLE,diagnosed according to American College of Rheumatology(ACR)revised classification criteria.18 SLE patients with clinical suspicion of were collected.The concentration of 25(OH)D of these patients and the expression of TLR 9 by immunohistochemistry.Results:1.The difference in comparison of TLR-9 mRNA of three groups before intervention has no statistical significance(P>0.05);there was no obvious change in control group after intervention,while the TLR-9 mRNA of BLyS blocker group and TLR-9 blocker group have decreased distinctly,and TLR-9 mRNA of TLR-9 blocker group was lower than that of BLyS blocker group,with statistical significance(P<0.05).The difference in comparison of BLyS protein concentration and IL-10 level of three groups before intervention has no statistical significance(P>0,05);there was no obvious change in control group after intervention,while BLyS protein concentration and IL-10 level of BLyS blocker group and TLR-9 blocker group have decreased distinctly,with statistical significance(P<0.05);but the difference in comparison between BLyS blocker group and TLR-9 blocker group has no statistical significance(P>0.05).The difference in comparison of anti-dsDNA antibody titer and C3,C4,ESR,CRP level of three groups before intervention has no statistical significance(P>0.05);there was no obvious change in control group after intervention,while above indexes of BLyS blocker group and TLR-9 blocker group have decreased distinctly,with statistical significance(P<0.05);but the difference in comparison between BLyS blocker group and TLR-9 blocker group has no statistical significance(P>0.05).2.Compared with SLE patients in stable condition,the ones with active situation,the expression of TLR9 mRNA,IL-10 was significantly higher(P<0.01),while 25(OH)D levels decreased significantly(P<0.01).3.LN sections showed a high amount of TLR9 expression.We found a positive correlation between TLR9 and disease activity.We detected a negative correlation between glomerular TLR9 and the concentration of 25(OH)D(r=-0.764/P<0.01),as well as tubular TLR9(r=-0.741 p<0.01).We also detected a positive correlation between tubular TLR9 expression and SLEDAI-INDEX(r=0.61 p=0.008),as well as tubular TLR9(r=0.61 p=0.007).Conclusion:1.It implied that TLR-9 may be an important signaling pathway of BLyS induced SLE,regulating inflammatory immune level,so taking intervention for TLR-9 or BLyS expression can inhibit autoimmune injury process of SLE.2.The increased expression of TLR9 mRNA and IL-10 implied the increased activity of the disease.3.1,25(OH)2 D3 may dcrease the expression of TLR9 and play a role in the treatment of SLE. |
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