摘要
目的:探讨高迁移率族蛋白1(HMGB1)对呼吸道合胞病毒(RSV)毛细支气管炎合并肾脏受累型过敏性紫癜(HSP)患儿单核细胞免疫功能的影响。方法:选择初诊为RSV毛细支气管炎合并肾脏受累HSP患儿30例作为观察组,选取同期进行体检的健康足月婴幼儿30名作为对照组,2组婴幼儿入院后采集血浆样本,进行常规血细胞分析和C反应蛋白(CRP)水平检测,并采用ELISA法检测2组婴幼儿血浆中HMGB1的表达水平。采用人重组RSV-A2病毒转染人支气管上皮细胞系16HBE,并收集细胞培养上清检测HMGB1。使用经孔板共培养RSV感染细胞、无感染组的人支气管上皮细胞系16HBE和单核细胞THP1,qRT-PCR法测定细胞总RNA,采用Western blotting检测单核细胞中Toll样受体TLR-4和TLR-7的水平。结果:RSV毛细支气管炎合并肾脏受累HSP患儿外周血较健康对照组较健康对照组中HMGB1的表达水平升高(t=5.84,P<0.05),且外周血单核细胞数量、外周淋巴细胞数量及CRP水平与RSV毛细支气管炎合并肾脏受累HSP患儿外周血中HMGB1表达水平呈正相关关系(r=0.606、0.301、0.394,P<0.05),其中外周血单核细胞数量与外周血中HMGB1表达水平相关性最显著(P<0.05)。经qRT-PCR检测,经病毒转染后的16HBE细胞培养基中HMGB1的表达相较对照组显著增加(t=5.41,P<0.05)。RSV转染的16HBE细胞、感染对照组、HMGB1抑制组分别与单核细胞THP-1进行共培养后,经Western blotting检测单核细胞中TLR-4和TLR-7蛋白的表达水平,结果显示THP-1细胞中TLR-4的表达水平增加(P<0.05),而TLR-7水平无显著变化(P>0.05)。结论:HMGB1的表达水平与儿童RSV毛细支气管炎合并肾脏受累HSP病情的发展有关,HMGB1在单核细胞介导的免疫性炎症中起着重要作用,对掌握儿童RSV毛细支气管炎的发病机制具有重要意义。
Objective:To investigate the effects of high mobility group protein 1(HMGB1)on monocyte immune function in children with respiratory syncytial virus(RSV)bronchiolitis and henoch-schonlein purpura(HSP).Methods:A total of 30 hospitalized children with RSV bronchiolitis complicated with HSP were selected as the observation group,and 30 healthy full-term infants who underwent physical examination during the same period were selected as the control group.Plasma samples were collected from the two groups after admission,the routine blood cell analysis and C reactive protein(CRP)level detection were performed,and the serum expression levels of HMGB1 in two groups were detected using ELISA.The human bronchial epithelial cell line 16HBE was transfected with recombinant RSV-A2 virus,and the HMGB1 level in cell culture supernatant was detected.RSV-infected cells,human bronchial epithelial cell line 16HBE and monocyte THP1 were co-cultured by hole plates,the total RNA of cells was determined by qRT-PCR,and the expression levels of Toll-like receptor TLR-4 and TLR-7 of monocytes were detected using Western blotting.Results:Compared with the healthy control group,the serum expression level of HMGB1 in RSV bronchiolitis children complicated with HSP was significantly higher(t=5.84,P<0.05).The number of monocytes,peripheral lymphocytes and serum CRP level were positively correlated with the serum level of HMGB1 in RSV bronchiolitis children complicated with HSP(r=0.606,0.301,0.394,P<0.05).The number of monocytes in peripheral blood was the most significantly correlated with serum level of HMGB1(P<0.05).The result of qRT-PCR showed that the HMGB1 level in 16HBE cell culture medium after virus transfection significantly increased compared with that in control group(t=5.41,P<0.05).After co-culture of RSV-transfected 16HBE cells,infection control group and HMGB1 inhibited group with monocyte THP-1,the expression levels of TLR-4 and TLR-7 proteins in monocytes were detected by Western blotting,and the results of which showed that the expression level of TLR-4 in THP-1 cells significantly increased(P<0.05),while the change of TLR-7 level was not significant(P>0.05).Conclusions:The expression level of HMGB1 is related to the development of RSV bronchiolitis complicated with HSP.HMGB1 plays an important role in monocyte-mediated immune inflammation,which is of great significance to understand the pathogenesis of pediatric RSV bronchiolitis.
作者
王银洁
蔡花
张菂
黄翠影
陈瑞珊
王彩芳
WANG Yinjie;CAI Hua;ZHANG Di;HUANG Cuiying;CHEN Ruishan;WANG Caifang(Department of Critical Care Medicine,Kailuan General Hospital,Tangshan Hebei 063000,China)
出处
《蚌埠医学院学报》
CAS
2024年第6期758-762,共5页
Journal of Bengbu Medical College
基金
河北省卫健委重点科技研究计划(20191335)。
