| BackgroundIdiopathic nephrotic syndrome?INS?is the most common glomerular disease in childhood with age between 2 and 8 years old.The managentment of INS always faces with challenges of repeated infection,frequently relapsing nephrotic syndrome,steroid-dependent and steroid-resistent nephrotic syndrome,however,the pathogenesis of INS still remains unclear.The efficacy of immunosuppressive treatments indicates that INS may be associated with dysfunctions of the immune system.Recognition of pathogen-associated molecular patterns?PAMPs?by T cells bearing Toll-like receptors?TLRs?induces their activation and the secretion of proinflammatory cytokines.TLRs also induce B cell differentiation and Ig production via nuclear factor-kappa B?NF-?B?in a T cell-independent manner.Recognition of PAMPs by TLRs triggers the NF-?B,activator protein 1?AP-1?and interferon?IFN?pathways,resulting in the translocation of transcription factors and cytokine modulation leading to inflammatory responses via T cell stimulation.TLRs can provide constimulatory signals for acquired immune system,and eventually,induce a series of immune-mediated inflammatory response.Dysfunction of ILRs?such as TLR-2,4,7 etc.?is involved in many kidney diseases.But it is not clear whether TLR-1,3,9 is involved in the pathogenesis of INS in children.So it is necessary to explore the pathogenesis of TLR-1,3,9 in children INS.ObjectiveWe hypothesize that TLRs in PBMC might be involved in INS disease.The aim of this study was to investigate the expression and function of TLR-1,3,9 in PBMC of INS patients at various stages of the disease,and to explore the possible roles of TLR-1,TLR-3and TLR-9 in the pathogenesis of INS in children.MethodsPeripheral venous blood was collected from the INS children and healthy control children with empty stomach in the early morning.PBMC was separated by Ficoll-hypzue and the total RNA was extracted by RNA extraction kit.The real-time polymerase chain reaction?RT-PCR?was used to detect the expressions of TLR-1mRNA,TLR-3mRNA and TLR-9mRNA in 45 children with INS and 30 healthy children?normal control group?.Meantime,the relation of TLR-1mRNA,TLR-3m RNA and TLR-9mRNA and serum albumin and cholesterol levels were confirmed.ResultsThere were no significant difference in age and sex between experimental group and control group(t=-1.660,x?17?=0.009,P all>0.05).The expression of TLR-1mRNA/?-actin in PBMC of children with INS before treatment?activity group?was significantly higher than that in remission group?2.432±0.231 vs 1.675±0.627,t=7.599,P<0.05?and control group?2.432±0.231 vs 0.512±0.228,t=35.446,P<0.05?,and the differences were statistically significant in the expression of TLR-1mRNA/?-actin between remission group and control group?1.675±0.627 vs0.512±0.228,t=9.722,P<0.05?.The expression of TLR-3 mRNA/?-actin in PBMC of children with INS before treatment?activity group??0.987±0.124?was significantly higher than that in remission group?0.501±0.016?and control group?0.021±0.001?,the differe-nces were statistically significant?t=26.076,42.571,P all<0.05?,and the remission group was significantly higher than that in control group?t=163.732,P<0.05?.The expression of TLR-9 mRNA/?-actin in PBMC of children with INS before treatment?activity group??1.965±0.952?was significantly higher than that in remission group?1.336±0.282?and control group?0.790±0.731?,the differences were statistically significant?t=4.249,5.734,P all<0.05?,and the remission group was significantly higher than that in control group?t=4.541,P<0.05?.The serum levels of albumin in children with INS before treatment?activity group??23.62±11.67?g/L was significantly lower than that in remission group?42.19±16.3?g/L and control group?46.88±14.80?g/L,the differences were statistically significant?t=-6.20,-7.58,P all<0.05?.The serum levels of cholesterol in children with INS before treatment?activity group??9.54±2.53?mmol/L was significantly higher than that in remission group?3.01±1.72?mmol/L and control group?2.89±1.66?mmol/L,the differences were statistically significant?t=14.32,12.677,P all<0.01?.The serum levels of albumin and cholesterol in children with INS after treatment were not statistically significantly difference compared with the those of the control group?t=-1.264,0.30,P all>0.05?.The correlation analysis showed that there was a negative correlation between the expressions of TLR-1mRNA,TLR-3m RNA and TLR-9m RNA and the serum level of albumin in PBMC of children with INS before treatment?r=-0.457,-0.891,-0.125,respectively,P all<0.05?.However there was a positive correlation between the expressions of TLR-1mRNA,TLR-3m RNA and TLR-9m RNA and the serum level of cholesterol?r=0.445,0.911,0.872,P all<0.05?.Conclusions1.TLR-1mRNA,TLR-3mRNA and TLR-9mRNA in PBMC of children with INS were significantly higher,suggesting that the abnormal activation of TLR-1,TLR-3 and TLR-9 may participate in the pathogenesis of INS.2.TLR-1mRNA,TLR-3mRNA and TLR-9mRNA were correlated with the serum level of the albumin and cholesterol in children with INS,suggesting that the TLR-1,TLR-3 and TLR-9 may be related to the activity of INS. |
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