CAJ | 학술논문

目的研究过敏性紫癜(HSP)患儿急性期外周血单核细胞(PBMC)TLR2,4,6的表达及其与血浆白细胞介素(IL)-17的相关性,探讨TLR2,4,6在 HSP 发病机制中的作用。方法选取2013年4月至12月青岛大学附属医院收治入院的 HSP 急性期患儿42例为研究对象(纳入 HSP 组),年龄为4~13岁,男性患儿为25例,女性为17例。另选取同期于本院儿童保健科进行健康体检的30例儿童纳入对照组。采用流式细胞术检测 HSP 患儿 PBMC TLR2,4,6蛋白表达情况,采用实时荧光定量 PCR(RT-PCR)法检测 HSP患儿PBMC髓样分化蛋白88(MyD88)mRNA的相对表达水平,采用酶联免疫吸附测定(ELISA)法检测血浆白细胞介素(IL)-17水平。组间比较采用独立样本t检验,相关性分析采用Pearson相关检验。本研究遵循的程序符合青岛大学附属医院人体试验委员会所制定的伦理学标准,得到该委员会批准,并征得受试对象监护人的知情同意,并与之签署临床研究知情同意书。结果①HSP 患儿 PBMC TLR2,4,6蛋白表达均显著高于对照组,差异均有统计学意义(t=9.432,P<0.01；t=8.876,P<0.01；t=10.486,P<0.01)。②HSP 组 MyD88 mRNA 相对表达水平(1.279±0.419)较对照组水平(0.995±0.251)显著升高,二者比较,差异有统计学意义(t=2.463,P<0.05)。③HSP组患儿血浆 IL-17水平较对照组显著增高[(13.168±2.841)pg/mL vs (11.091±1.768)pg/mL],差异有统计学意义(t=2.641,P<0.05)。④HSP组PBMC TLR2,4,6蛋白表达均与MyD88 mRNA的相对表达水平呈显著正相关关系(r=0.850,P<0.01；r=0.669,P<0.01；r=0.788,P<0.01)；与血浆IL-17水平均呈正相关关系(r=0.438,P<0.01；r=0.404,P<0.05；r=0.357,P<0.05)。结论 TLR2,4,6信号途径的异常活化可能参与 HSP发生发展,活化的TLR2,4,6可能通过上调辅助性T细胞(Th17)介导 HSP的免疫发病机制。
목적연구과민성자전(HSP)환인급성기외주혈단핵세포(PBMC)TLR2,4,6적표체급기여혈장백세포개소(IL)-17적상관성,탐토TLR2,4,6재 HSP 발병궤제중적작용。방법선취2013년4월지12월청도대학부속의원수치입원적 HSP 급성기환인42례위연구대상(납입 HSP 조),년령위4~13세,남성환인위25례,녀성위17례。령선취동기우본원인동보건과진행건강체검적30례인동납입대조조。채용류식세포술검측 HSP 환인 PBMC TLR2,4,6단백표체정황,채용실시형광정량 PCR(RT-PCR)법검측 HSP환인PBMC수양분화단백88(MyD88)mRNA적상대표체수평,채용매련면역흡부측정(ELISA)법검측혈장백세포개소(IL)-17수평。조간비교채용독립양본t검험,상관성분석채용Pearson상관검험。본연구준순적정서부합청도대학부속의원인체시험위원회소제정적윤리학표준,득도해위원회비준,병정득수시대상감호인적지정동의,병여지첨서림상연구지정동의서。결과①HSP 환인 PBMC TLR2,4,6단백표체균현저고우대조조,차이균유통계학의의(t=9.432,P<0.01；t=8.876,P<0.01；t=10.486,P<0.01)。②HSP 조 MyD88 mRNA 상대표체수평(1.279±0.419)교대조조수평(0.995±0.251)현저승고,이자비교,차이유통계학의의(t=2.463,P<0.05)。③HSP조환인혈장 IL-17수평교대조조현저증고[(13.168±2.841)pg/mL vs (11.091±1.768)pg/mL],차이유통계학의의(t=2.641,P<0.05)。④HSP조PBMC TLR2,4,6단백표체균여MyD88 mRNA적상대표체수평정현저정상관관계(r=0.850,P<0.01；r=0.669,P<0.01；r=0.788,P<0.01)；여혈장IL-17수평균정정상관관계(r=0.438,P<0.01；r=0.404,P<0.05；r=0.357,P<0.05)。결론 TLR2,4,6신호도경적이상활화가능삼여 HSP발생발전,활화적TLR2,4,6가능통과상조보조성T세포(Th17)개도 HSP적면역발병궤제。
Objective To study the expression of TLR2,4,6 protein in peripheral blood mononuclear cell (PBMC)and the relationship with helper T cell (Th)17 of children with Henoch-Schonlein purpura (HSP),and to investigate the roles of the TLR2,4,6 in the pathogenesis of HSP.Methods Forty-two children with acute phrase HSP who were hospitalized from April 2013 to December 2013 in the Affiliated Hospital of Qingdao University were enrolled (HSP group),aged 4-13.Among them,there were 25 males and 17 females.At the same time,another 30 healthy children from Child Health Division of the same hospital were served as control group.TLR2,4,6 protein expression levels were detected by flow cytometric analysis.MyD88 mRNA expression levels in PBMC were detected by real-time fluorescent polymerase chain reaction (RT-PCR).The levels of plasma interleukin (IL)-1 7 were detected by enzyme linked immunosorbent assay (ELISA).In this study,t test and Pearson correlation test of statistical analysis were used between two groups.The study protocol was approved by the Ethical Review Board of Investigation of Affiliated Hospital of Qingdao University. Informed consent was obtained from all participates′parents.Results ①Compared with control group,the expression level of TLR2,4,6 protein were remarkably increased in HSP group (t=9.432,P<0.01;t=8.876,P<0.01;t=10.486,P<0.01).② The level of MyD88 mRNA in HSP group (1.279±0.419)was higher than that of control group (0.995±0.251)with significant difference (t=2.463,P<0.05).③The levels of plasma IL-17 of HSP group was significantly higher than that of control group [(13.168±2.841)pg/mL vs (11.091±1.768) pg/mL]with significant difference (t=2.641,P<0.05).④ The TLR2,4,6 protein expression of PBMC and the level of MyD88 mRNA expression of HSP group showed a significant positive correlation (r=0.85, P<0.01;r=0.669,P<0.01;r=0.788,P<0.01);and its also had positive correlation with the level of plasma IL-17.Conclusions The aberrant activation of the TLR2,4,6 may participate in the pathogenesis of HSP.The hyper-activation of TLR2,4,6 might upregulate the expression Th17 and mediate the immunology mechanism.