| Part I Diffrential expression of TLR/My D88 signaling pathway is associated with microscopic polyangiitis in peripheral blood mononuclear cells? Objective ? To identify the differentially expressed genes of TLR/My D88-related gene m RNA expression levels in peripheral blood mononuclear cells(PBMC)of active stage patients with microscopic polyangiitis(MPA)and healthy volunteers.?Methods?A case-control study was performed among 10 active stage patients with MPA(case group)and 10 healthy volunteers(control group)and collect the the data of clinical manifestations.The total RNA was extracted from peripheral blood mononuclear cells of the collected blood samples and reversely transcribed in c DNA.A gene array(Human TLR for Autoimmunity &Inflammation Gene Array)analysis was performed to study TLR/My D88-related gene m RNA expression levels.We then used quantitative real-time q PCR(RT-PCR)to validate the array test.?Results?(1)The active stage patients with MPA was compared to the healthy volunteers,the expression profiles of 23 of the 84 genes showed significant change.Thirteen genes were up-regulated,up-regulated genes encompassed IRAK3(1.5 times,P=0.0205),TLR4(1.57 times,P=0.0165),TOLLIP(1.65 times,P=0.0191),CXCL11(1.67 times,P=0.0461),FADD(1.83 times,P=0.0239),TLR6(1.93 times,P=0.0213),IRF7(1.97 times,P=0.0498),CD14(2.02 times,P=0.0454),BCL3(2.02 times,P=0.0312),TLR1(2.42 times,P=0.0280),CXCL9(2.51 times,P=0.0498),MUC1(3.11 times,P=0.0132),IL10(10.17 times,P=0.0367),the difference expression of MUC1(3.11 times,P=0.0132)and IL10(10.17 times,P=0.0367)is more apparent.Ten genes were down-regulated,down-regulated genes encompassed CXCL10(-1.51 times,P=0.0404),SARM1(-1.56 times,P=0.0121),TLR10(-1.57 times,P=0.0499),ATF3(-1.58 times,P=0.0034),CIITA(-1.69 times,P=0.0237),JUN(-1.86 times,P=0.0127),RIPK2(-1.91 times,P=0.0131),CHUK(-15.26 times,P<0.0001),IFNA7(-16.13 times,P<0.0001),SOCS1(-22.35 times,P<0.0001),the difference expression of CHUK(-15.26 times,P<0.0001),IFNA7(-16.13 times,P<0.0001)and SOCS1(-22.35 times,P<0.0001)is more apparent.(2)The data of real-time q PCR Six genes were randomly selected from the 23 differential expression gene for validation.The fold change of the gene in the RT-PCR was as follows: MUC1(2.78 times,P=0.0392),BCL3(1.99 times,P=0.0138),IRF7(1.88 times,P=0.0116),IL10(2.91 times,P=0.0125),TLR10(-1.86 times,P=0.0203),JUN(-1.81 times,P=0.0408).The resulting RT-q PCR was overall consistent with the findings in gene array.? Conclusion ? In conclusion,our study showed the significant differentially expression levels of multiple genes related to TLR/My D88 signaling pathway in PBMC of active stage patients with MPA.These genes include toll-like receptors,negative regulation molecules,adaptors and TLR interacting proteins,effectors,TLR downstream pathway and target genes,activating transcription factors and inflammatory cytokines.TLR signaling pathway may be an important immune signaling pathway in the pathogenesis of MPA.Part II Differential expression of TLR/My D88 signaling pathway is associated with microscopic polyangiitis in peripheral blood Neutrophils? Objective ? To identify the differentially expressed genes of TLR/My D88-related gene m RNA expression levels in peripheral blood neutrophils of active stage patients with microscopic polyangiitis(MPA)and healthy volunteers.?Methods?A case-control study was performed among 20 active stage patients with MPA(case group)and 12 healthy volunteers(control group)and collect the the data of clinical manifestations.The total RNA was extracted from peripheral blood neutrophils of the collected blood samples and reversely transcribed in c DNA.A gene array(Human TLR for Autoimmunity &Inflammation Gene Array)analysis was performed to study TLR/My D88-related gene m RNA expression levels.We then used quantitative real-time q PCR(RT-PCR)to validate the array test.?Results?(1)The active stage patients with MPA was compared to the healthy volunteers,the expression profiles of 18 of the 84 genes showed significant change.Thirteen genes were up-regulated,up-regulated genes encompassed TRAF3(1.52 times,P=0.0237),NF?B1(1.65 times,P=0.0086),LY96(1.85 times,P=0.0002),IRF7(1.93 times,P=0.0188),IRAK4(1.93 times,P=0.0224),TLR6(2.00 times,P=0.0092),CASP1(2.01 times,P=0.0032),TBK1(2.06 times,P=0.0145),IRAK3(2.50 times,P=0.0066),TLR10(2.55 times,P=0.0048),IL10(2.70 times,P=0.008),JUN(2.90 times,P=0.0006),MAPK14(3.06 times,P=0.0015),the difference expression of MAPK14(3.06times,P=0.0015)is the most apparent.Five genes down-regulated genes encompassed TNRC5(-1.53 times,P=0.0003),TLR7(-1.85 times,P=0.0284),SIGIRR(-2.10 times,P=0.0227),CIITA(-2.26 times,P=0.0205),MAP3K7IP1(-2.28 times,P<0.0001),the difference expression of MAP3K7IP1(-2.28 times,P<0.0001)is the most apparent.(2)The data of real-time q PCR Four genes were randomly selected from the 18 differential expression gene for validation.The fold change of the gene in the RT-PCR was as follows: IL10(1.80 times,P=0.0478),TLR10(2.50 times,P=0.0224),NF-?B1(5.27 times,P=0.0057),TLR7(-10.65 times,P=0.0027).The resulting RT-q PCR was overall consistent with the findings in gene array.? Conclusion ? In conclusion,our study showed the significant differentially expression levels of multiple genes related to TLR/My D88 signaling pathway in peripheral blood neutrophils of active stage patients with MPA.These genes include toll-like receptors,negative regulation molecules,apoptosis-related genes,TLR interacting proteins,TLR downstream pathway and target genes,activating transcription factors and TLR accessory molecules.TLR signaling pathway may be an important immune signaling pathway in the pathogenesis of MPA. |
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