CAJ | 학술논문

目的：研究Toll样受体（Toll-like receptor，TLR）2、TLR4在过敏性紫癜（HSP）患儿外周血单核细胞的表达及其与Th1和Th2型免疫应答的相关性，探讨TLR2、TLR4在HSP发病机制中的作用。方法选取2011年10月-2012年11月于我院儿内科收治的HSP患儿64例为研究对象，分为HSP无肾损害组（NHSPN，36例）及HSP有肾损害组（HSPN，28例），另选取我院儿保科健康查体儿童30例作为正常对照组。采用流式细胞术检测外周血T淋巴细胞亚群及单核细胞TLR2及TLR4蛋白表达，实时荧光定量PCR技术检测外周血单核细胞TLR2 mRNA及TLR4 mRNA的基因相对表达量，ELISA法检测血浆IFN-γ、IL-4、IL-6水平。结果（1） HSP 患儿外周血单核细胞TLR2 mRNA、TLR4 mRNA相对表达量及TLR2、TLR4蛋白表达均显著高于正常对照组（P均＜0．01），HSPN组外周血单核细胞TLR2 mRNA、TLR4 mRNA相对表达量及TLR2、TLR4蛋白表达均明显高于NHSPN组（P＜0．05；P＜0．01；P＜0．01；P＜0．01）。（2）HSP组CD3＋T细胞、CD3＋CD4＋T细胞显著降低，CD3＋CD8＋T细胞及CD3＋HLADR＋活化T细胞明显升高（P均＜0．01）。（3）HSP组血浆IL-4、IL-6水平显著高于正常对照组（P＜0．01，P＜0．01），IFN-γ水平显著低于正常对照组（P＜0．05），IFN-γ/IL-4比值显著低于对照组（P＜0．01）。（4）HSP组TLR2、TLR4蛋白表达与血浆IL-4、IL-6水平均呈显著正相关（P＜0．01，P＜0．05；P＜0．01，P＜0．01）；与IFN-γ/IL-4比值均呈显著负相关（P＜0．01；P＜0．01）。结论TLR2及TLR4活化可能参与了HSP的免疫发病机制，TLR2及TLR4的过度活化可能与HSP的肾损伤有关。 HSP患儿存在T淋巴细胞亚群失调，功能紊乱，Th1/Th2失衡。活化的TLR2及TLR4可能通过上调Th2型免疫应答介导HSP的免疫发病机制。
목적：연구Toll양수체（Toll-like receptor，TLR）2、TLR4재과민성자전（HSP）환인외주혈단핵세포적표체급기여Th1화Th2형면역응답적상관성，탐토TLR2、TLR4재HSP발병궤제중적작용。방법선취2011년10월-2012년11월우아원인내과수치적HSP환인64례위연구대상，분위HSP무신손해조（NHSPN，36례）급HSP유신손해조（HSPN，28례），령선취아원인보과건강사체인동30례작위정상대조조。채용류식세포술검측외주혈T림파세포아군급단핵세포TLR2급TLR4단백표체，실시형광정량PCR기술검측외주혈단핵세포TLR2 mRNA급TLR4 mRNA적기인상대표체량，ELISA법검측혈장IFN-γ、IL-4、IL-6수평。결과（1） HSP 환인외주혈단핵세포TLR2 mRNA、TLR4 mRNA상대표체량급TLR2、TLR4단백표체균현저고우정상대조조（P균＜0．01），HSPN조외주혈단핵세포TLR2 mRNA、TLR4 mRNA상대표체량급TLR2、TLR4단백표체균명현고우NHSPN조（P＜0．05；P＜0．01；P＜0．01；P＜0．01）。（2）HSP조CD3＋T세포、CD3＋CD4＋T세포현저강저，CD3＋CD8＋T세포급CD3＋HLADR＋활화T세포명현승고（P균＜0．01）。（3）HSP조혈장IL-4、IL-6수평현저고우정상대조조（P＜0．01，P＜0．01），IFN-γ수평현저저우정상대조조（P＜0．05），IFN-γ/IL-4비치현저저우대조조（P＜0．01）。（4）HSP조TLR2、TLR4단백표체여혈장IL-4、IL-6수평균정현저정상관（P＜0．01，P＜0．05；P＜0．01，P＜0．01）；여IFN-γ/IL-4비치균정현저부상관（P＜0．01；P＜0．01）。결론TLR2급TLR4활화가능삼여료HSP적면역발병궤제，TLR2급TLR4적과도활화가능여HSP적신손상유관。 HSP환인존재T림파세포아군실조，공능문란，Th1/Th2실형。활화적TLR2급TLR4가능통과상조Th2형면역응답개도HSP적면역발병궤제。
Objective To explore the role of TLR2 and TLR4 in the pathogenesis of Henoch-Schonlein purpura ( HSP) by investigating their expression at mRNA and protein levels in peripheral blood mononuclear cells ( PBMCs ) and their influences on Th 1/Th2 immune response in children with HSP . Methods 64 hospitalized children with HSP in the Affiliated Hospital of Qingdao University Medical Col -lege from October 2011 to November 2012 were enrolled in the study .They were further divided into non-He-noch-Schonlein purpura nephritis ( NHSPN ) group ( n =36 ) and Henoch-Schonlein purpura nephritis (HSPN) group (n=28).30 age-matched healthy children from Child Health Division of the same hospital were selected as controls .The expression of TLR2 and TLR4 at mRNA level in PBMCs were detected by re-al-time fluorescent polymerase chain reaction .The expression of TLR2 and TLR4 at protein level and T cells subset were detected by flow cytometry .The levels of IFN-γ, IL-4 and IL-6 in plasma were determined by enzyme-linked immunosorbent assay (ELISA).Results (1)Compared with the control group , the expres-sion of TLR2 and TLR4 at mRNA and protein levels were remarkably increased in children with HSP , espe-cially in HSPN group.(2)Compared with the control group, the percentage of CD3+T cells and CD3+CD4+T cells were down-regulated in HSP group , but the percentage of CD 3+CD8+T cells and CD3+HLADR+T cells were up-regulated.(3)The level of IFN-γand the ratio of IFN-γ/IL-4 in plasma from children with HSP were significantly lower than those of the controls , while the level of IL-4 and IL-6 were remarkably higher than those of the controls .(4)The expression of TLR2 and TLR4 at protein level in PBMCs from chil-dren with HSP showed significant positive correlations with the expression of TLR 2 and TLR4 at mRNA level and plasma concentration of IL-4 and IL-6, but a negative correlation with the ratio of IFN-γ/IL-4.Conclu-sions The aberrant activation of TLR 2 and TLR4 might be correlated with the immunological pathogenesis of HSP by enhancing Th2 immune response.The hyper-activation of TLR2 and TLR4 might result in renal injury in patients with HSP .