【作者】 余强；

【导师】 傅睿；

【作者基本信息】 南昌大学 ， 儿科学， 2013， 硕士

【摘要】 实验目的：本研究旨在检测各临床类型过敏性紫癜(HSP)患儿不同治疗时期血清中IgA1异常糖基化与肝细胞生长因子(HGF)的水平，及HSP患儿胃粘膜和肾脏组织中IgA、IgA1、HGF的表达情况，综合分析它们与HSP疾病不同时期以及相关脏器损伤的关系，以进一步探讨IgA1与HGF在HSP疾病发生发展过程中的作用机制，并对临床诊断、鉴别诊断和治疗提供新思路。实验研究方法：1、研究对象为2011年7月至2012年11月我院肾内科收治的72例HSP初发急性期住院患儿，病例均符合2005年欧洲风湿病防治委员会和儿童肾病防治委员会共同制订的HSP新诊断标准，并排除有重症感染、严重的心脏、肝脏、血液系统性疾病，以及长期服用肾上腺皮质激素、免疫抑制剂等病例。2、分组将HSP患儿按不同治疗时期及临床表现分为8组：单纯组20例，腹痛组20例，过敏性紫癜(HSPN)组20例，消化道出血组12例，以及与以上四组对应的病情缓解组；诊断非HSP的急性胃肠炎或消化性溃疡患儿20例及儿童保健门诊体检的健康儿童20例作为对照组；HSP患儿胃粘膜组织20例，急性胃肠炎患儿胃粘膜组织10例及胃肿瘤患儿远端正常胃粘膜组织4例作为对照组；HSPN患儿肾脏组织40例，肾脏肿瘤患儿远端正常肾组织10例作为正常对照组。3、观察指标：血清IgAl-蚕豆凝集素(VVL)结合实验方法、ELISA法检测各临床类型HSP患儿急性期与缓解期及对照组血清IgA1异常糖基化和HGF的水平；免疫组织化学非生物素二步法测定IgA、IgA1、HGF在胃粘膜、肾脏组织中的表达情况；间接免疫荧光法观察IgA1在肾脏组织中的表达情况。结果：1、HSP患儿血清IgA1异常糖基化水平(mg/L)：①急性期单纯组(3.38±1.35)、腹痛组(3.47±1.59)、HSPN组(5.06±0.96)及消化道出血组(3.90±1.06)之间比较，HSPN组明显高于其它三组，但HSP四组均高于非HSP胃肠炎组(2.53±1.06)及正常对照组(2.51±0.83)，差异有统计学意义(P<0.05)；②缓解期单纯组(2.45±1.29)、腹痛组(2.37±1.38)及消化道出血组(2.60±1.00)较急性期明显下降(P<0.05)，但HSPN组(4.66±1.38)与急性期比较差异无统计学意义(P>0.05)；2、HSP患儿血清HGF水平(ng/L)：①急性期单纯组(2.86±1.31)、腹痛组(2.57±1.90)、HSPN组(3.91±0.91)及消化道出血组(3.85±0.71)均明显高于正常对照组(1.04±0.55)，组间比较HSPN组及消化道出血组高于其他两组，差异有统计学意义(P<0.05)，与非HSP胃肠炎组(2.28±1.26)比较，仅HSPN组和消化道出血组高于非HSP胃肠炎组(P<0.01)，其余两组与其比较差异均无统计学意义(P>0.05)；②缓解期单纯组(1.21±1.26)、腹痛组(1.37±0.87)、HSPN组(1.91±0.76)及消化道出血组(1.84±1.24)较急性期明显下降，但HSPN组及消化道出血组仍高于其他两组，差异有统计学意义(P<0.05)；③HSP患儿血清IgA1异常糖基化水平与HGF水平之间存在正相关关系(P<0.05)；3、HSP患儿免疫组化结果，观察到均有IgA、IgA1、HGF蛋白在胃粘膜及肾脏组织中大量沉积，表达水平显著高于非HSP胃肠炎组及正常对照组，差异有统计学意义(P<0.05)，且相同组织中IgA1与HGF表达呈正相关关系(P<0.05)。结论：（1）HSP各临床类型急性期血清IgA1异常糖基化及HGF水平显著高于非HSP胃肠炎组及正常对照组，且缓解期较急性期有所下降，提示异常糖基化的IgA1及HGF在HSP的发生、发展中起到一定作用。（2）在腹型HSP患儿胃粘膜组织及HSPN患儿肾脏组织中均可以观察到IgA、IgA1的大量沉积，且IgA1/IgA免疫分数比值明显增高，远高于非HSP胃肠炎组，结合HSP患儿血清IgA1异常糖基化水平的升高，提示异常糖基化的IgA1沉积参与了HSP胃粘膜及肾脏组织的损伤过程；HGF不但在胃粘膜及肾脏组织中大量沉积，且与组织中的IgA1呈正相关关系，提示HGF可能在HSP胃粘膜及肾脏受损过程中起到拮抗IgA1的作用，并且对于无典型皮疹的腹型HSP患儿，IgA1及HGF可能作为新的病理学指标对HSP诊断及鉴别提供新的依据。更多还原

【Abstract】 Objective:To investigate the relation of aberrant glycosylation of IgA1andhepatocyte growth factor(HGF) to different stage and visceral organ injure inchildren Hen ch-Sch nlein purpura(HSP) through detecting the concentration of inserum of in children HSP in acute-stage and remittent-stage，and observing theexpression of IgA,IgA1,HGF in gastrointestinal tract and renal.To comprehensiveanalysis those two factors’ effect on the pathogenesis of HSP,and to provide newideas for clinical diagnosis,differential diagnosis,remedy of HSP.Methods:1.72children with HSP in nitialdose happen symptom and at acute stagewho were in Nephrology department of Children Hospital from2011Jul to2012Nov,were enrolled in the study.The diagnosis of HSP was made according to theEuropean League against Rheumatism criteria in2005.None of patients had seriousinfection,severe disease of heart,kidney and blood system，received glucocoritcoidsand immunosuppressants.According to the clinical manifestation,all patients were classified into8groups:simple group(20children),abdominal group(20children),Hen ch-Sch nleinpurpura nephrology(HSPN)(20children),gastrointestinal tract hemorrhage group(20children),and the other groups were72children at remittent stage throughthe treatment;20children in acute gastroenteritis or peptic ulcer and to20he-althy children were divided into control group.The gastric mucosa of HSP(20children) were experimental group,non-HSP gastroenteritis(20children) and sto-mach neoplasm’s(4children) gastric mucosa were control group;meanwhile thekidney tissue of HSP(40children) were experimental group,and renal carcino-ma’s(10children) kidney tissue were control group.To detect the concentration of aberrant glycosylation of IgA1and HGF inserum of four HSP groups and controll gourps at the acute stage and the remittent stage through IgAl-Vicia Villosa Lectin hinge method and enzyme-labeledimmunosorbent assay method,and with immunohistochemistry method to observethe expression of IgA,IgA1,HGF in gastrointestinal tract and renal,in addition useindirect immunofluorescence to investigate the express of IgA1in renal.Results:1.The concentration of aberrant glycosylation of IgA1in serum(mg/L):at acutestage,compare with simple group(3.38±1.35),abdomina group(3.47±1.59),HSPNgroup(5.06±0.96),gastrointestinal tract hemorrhage group(3.90±1.06),HSPN groupwas all significantly higher than the other three groups,but four HSP groups were allhigher than non-HSP gastroenteritis group(2.53±1.06) and normal control(2.51±0.83)(P<0.05);at remittent stage,simple HSP group(2.45±1.29),abdominalgroup(2.37±1.38),gastrointestinal tract hemorrhage group(2.60±1.00) wereobviously lower than the acute stage,but HSPN group(4.66±1.38)was assame(P>0.05);2.The concentration of HGF in serum(ng/L):at acute stage,simple group(2.86±1.31),abdominal group(2.57±1.90),HSPN group(3.91±0.91),gastrointestinal tracthemorrhage group(3.85±0.71) were obviously higher than normal controlgroup(1.04±0.55),compare with four HSP groups,HSPN group and gastrointestinaltract hemorrhage group were higher than the other two groups,and also higher thannon-HSP gastroenteritis group(2.28±1.26)(P<0.05);at remittent stage,simplegroup(1.21±1.26),abdominal group(1.37±0.87),HSPN group(1.91±0.76),gastrointestinal tract hemorrhage group(1.84±1.24) were obviously lowerthan the acute stage，but HSPN group and gastrointestinal tract hemorrhage groupwere still higher than the other two groups(P<0.05);HSP children’s level of aberrantglycosylation IgA1and HGF were positive correlation(P<0.05);3.Aberrant glycosylation IgA1and HGF in HSP group were significantlyincreased than control group in gastrointestinal tract and renal(P<0.05),and aberrantglycosylation IgA1and HGF in the same tissue were positive correlation(P<0.05).Conclusion:1.The concentration of aberrant glycosylation of IgA1and HGF in serum of allkinds of HSP group obviously higher than non-HSP gastroenteritis group and normal control，however the remittent stage were lower than the acute stage，point out thataberrant glycosylation of IgA1and HGF play the important role in HSP,and thosefactors are maybe the new serum indexes of HSP for diagnosis,differential diagnosiswith other disease;2.The expression of IgA,IgA1were increased in gastrointestinal tract andrenal,and IgA1/IgA were more higher than non-HSP gastroenteritis group,with thelevel of aberrant glycosylation IgA1in serum,point out that aberrant glycosylationIgA1was participation in the process of gastrointestinal tract and renal injury inHSP;as the same,The expression of HGF were increased in gastrointestinal tract andrenal，and was positive correlation with IgA1,point out that HGF may be protect thetissue and antagonism the function of IgA1,in addition those two factors are maybethe new pathological indicators for diagnosis and differential diagnosis of HSP.更多还原