本文選題：IgA腎病 + 腭扁桃體��； 參考：《中南大學(xué)》2014年博士論文

【摘要】：研究背景：IgA腎病以IgAl/低糖基化IgAl沉積于腎小球系膜區(qū)及毛細(xì)血管袢為病理特征,以血尿、蛋白尿、高血壓及腎功能損害為主要臨床表現(xiàn),是世界范圍內(nèi)最常見(jiàn)的原發(fā)性腎小球疾病。大約20-40%的患者在診斷為IgA腎病的20年內(nèi)進(jìn)展至終末期腎功能衰竭,是導(dǎo)致終末期尿毒癥的最常見(jiàn)原因。 IgA腎病患者在黏膜組織,特別是上呼吸道和腸道黏膜感染后常出現(xiàn)陣發(fā)性肉眼血尿。既往也有研究IgA腎病患者在腭扁桃體摘除后出現(xiàn)一過(guò)性的尿檢惡化,同時(shí)檢測(cè)到血中IgAl一過(guò)性升高。即可以通過(guò)手術(shù)對(duì)腭扁桃體的牽拉、擠壓等刺激,進(jìn)行的腭扁桃體“激惹”試驗(yàn)。 B淋巴細(xì)胞刺激因子(B-cell-activation factor,BAFF),是腫瘤壞死因子(tumor necrosis factor,TNF)配體超家族的重要成員。BAFF通過(guò)對(duì)正常B細(xì)胞的成熟、增殖和分化的影響,對(duì)維持B淋巴細(xì)胞在體內(nèi)的發(fā)展和體內(nèi)的平衡起著重要的作用。但其臨床應(yīng)用還需要進(jìn)一步的研究。 IgA具有IgAl和IgA2兩種亞型,近年的研究證實(shí)低糖基化IgAl是IgA腎病的關(guān)鍵致病因子。IgAl與IgA2的主要區(qū)別點(diǎn)在于IgAl存在鉸鏈區(qū)。在IgAl鉸鏈區(qū),β1,3半乳糖轉(zhuǎn)移酶(C1GALT1)在其特異性分子伴侶(COSMC)的輔助下,通過(guò)UDP-半乳糖載體將半乳糖轉(zhuǎn)移至IgAl的p1,3位乙酰半乳糖胺殘基上,完成IgAl的糖基化修飾,而ST6GalNAcⅡ則將唾液酸轉(zhuǎn)至IgAl的α2,6位乙酰半乳糖胺殘基上,阻止了IgAl糖基化的修飾。IgA腎病外周血B淋巴細(xì)胞C1GALT1及COSMC活性明顯下降,并且與IgAl病理?yè)p傷嚴(yán)重程度呈正相關(guān),但是其活性下降的具體機(jī)制尚未闡明。既往研究發(fā)現(xiàn),外周血B淋巴細(xì)胞在無(wú)刺激時(shí)ST6GalNAcⅡmRNA的表達(dá)與正常對(duì)照組相比降低,另有研究發(fā)現(xiàn)IgAl分泌細(xì)胞在IL-4、IL-6的刺激下ST6GalNAcⅡmRNA顯著升高。 本課題中我們通過(guò)對(duì)IgA腎病患者以及對(duì)照組腭扁桃體摘除前后的研究,以腭扁桃體摘除術(shù)作為腭扁桃體“激惹”試驗(yàn),觀(guān)察腭扁桃體摘除前后外周血中的改變,分析其與外周血中B細(xì)胞活化、IgAl及其糖基化之間的關(guān)系,為IgA腎病的防治提供新的理論依據(jù)和治療靶點(diǎn)。 第一章IgA腎病患者腭扁桃體摘除前后外周血BAFF及IgAl比較 目的：檢測(cè)腭扁桃體摘除前后,外周血單個(gè)核細(xì)胞(PBMC)中BAFF以及血漿中IgAl表達(dá),分析腭扁桃體與B細(xì)胞活化及IgAl之間的關(guān)系。 方法：收集經(jīng)臨床和腎臟病理確診為IgA腎病患者在腭扁桃體摘除術(shù)術(shù)前(24-48h)術(shù)后(36-60h)的外周血16例做為IgA腎病組(IgAN組),行腭扁桃體摘除術(shù)的非IgA腎病患者術(shù)前術(shù)后外周血作為非IgA腎病組(non-IgAN組),以及正常對(duì)照組(healthy control組)16例。采用實(shí)時(shí)定量PCR(qRT-PCR)檢測(cè)BAFF在PBMC中mRNA的表達(dá)ELISA檢測(cè)血漿中IgAl含量。 結(jié)果： 1.qRT-PCR結(jié)果提示IgA腎病患者術(shù)前BAFF mRNA表達(dá)水平高于正常對(duì)照組(P0.01),IgA腎病患者術(shù)后BAFF表達(dá)較IgA腎病患者術(shù)前升高(P0.01),并高于非IgA腎病患者術(shù)后(P0.05)； 2.IgA腎病患者術(shù)前外周血血漿中IgAl的含量高于正常對(duì)照組(P0.05),IgA腎病患者術(shù)后血漿中IgAl的含量較術(shù)前升高(P0.01),并高于非IgA腎病患者術(shù)后(P0.05)； 3.相關(guān)性分析提示PBMC中BAFF mRNA表達(dá)與外周血IgAl含量呈正相關(guān)(r=0.604,P0.01)。 結(jié)論：本章實(shí)驗(yàn)通過(guò)模擬腭扁桃體感染,觀(guān)察到IgA腎病患者PBMC中BAFF基因表達(dá)增加,外周血血漿中IgAl含量升高與BAFF的關(guān)系密切,BAFF及其基因表達(dá)變化也有望成為一項(xiàng)評(píng)估患者IgA腎病病程變化的無(wú)創(chuàng)性檢測(cè)指標(biāo)。 第二章ST6GaINAcⅡ、C1GALT1及COSMC在外周血單個(gè)核細(xì)胞表達(dá)及與IgAl低糖基化相關(guān)性分析 目的：檢測(cè)腭扁桃體摘除前后,PBMC中ST6GalNAcⅡ, C1GALT1及COSMC表達(dá)以及外周血血漿中IgAl的低糖基化水平,分析腭扁桃體對(duì)低糖基化IgAl影響及IgAl低糖基化的調(diào)控機(jī)制。 方法：采用免疫組化方法檢測(cè)IgA腎病患者與非IgA腎病患者腭扁桃體中ST6GalNAcⅡ的表達(dá)；采用qRT-PCR檢測(cè)正常對(duì)照組、非IgA腎病組及IgA腎病組PBMC ST6GalNAcⅡ、C1GALT1及COSMCmRNA的表達(dá)；VVL-ELISA檢測(cè)外周血血漿中IgAl低糖基化水平。 結(jié)果： 1.免疫組化結(jié)果顯示ST6GalNAcⅡ在IgA腎病患者腭扁桃體中表達(dá)量高于非IgA腎病患者； 2.qRT-PCR結(jié)果提示IgA腎病患者術(shù)前ST6GalNAcⅡ mRNA表達(dá)水平高于正常對(duì)照組(P0.01),IgA腎病患者術(shù)后ST6GalNAcⅡ mRNA水平均較IgA腎病患者術(shù)前升高(P0.01),并高于非IgA腎病患者術(shù)后(P0.05)。IgA腎病患者術(shù)后36-48小時(shí)間,ST6GalNAcⅡ mRNA表達(dá)水平為術(shù)前的1.95倍(P0.01)；術(shù)后48-60小時(shí)間,ST6GalNAcⅡ mRNA表達(dá)水平為術(shù)前的1.38倍(P0.01)。 3.qRT-PCR結(jié)果提示IgA腎病患者術(shù)后C1GALT1、COSMCmRNA水平均較IgA腎病患者術(shù)前降低(P0.05)； 4.與正常對(duì)照組相比,IgA腎病組外周血血漿中的IgAl低糖基化程度明顯增高； 5.相關(guān)性分析提示PBMC中ST6GalNAcII mRNA表達(dá)與IgAl低糖基化水平呈正相關(guān)。 結(jié)論：IgA腎病患者腭扁桃體摘除術(shù)后,PBMC中ST6GalNAcII基因表達(dá)增加,C1GALT1、 COSMC基因表達(dá)減少,外周血血漿中Gd-IgAl含量升高,結(jié)合既往的研究,我們認(rèn)為PBMC中ST6GalNAcII活性的增高及C1GALT1/COSMC活性的降低,導(dǎo)致了對(duì)外周血中低糖基化IgAl分泌的增多。
[Abstract]:Background : IgA nephropathy is characterized by IgA1 / low glycosylated IgA1 deposited in glomerular mesangial area and capillary loop . It is the most common primary glomerular disease in the world . Approximately 20 - 40 % of patients progressed to end - stage renal failure during 20 years of diagnosis of IgA nephropathy , which is the most common cause of end - stage uremia .

IgA nephropathy patients often suffer from paroxysmal gross hematuria after mucosal tissue , especially in upper respiratory tract and intestinal mucosa .

B - cell - activation factor ( BAFF ) is an important member of tumor necrosis factor ( TNF ) ligand superfamily . BAFF plays an important role in the development of B - lymphocytes in vivo and in vivo .

IgA has two subtypes of IgA1 and IgA2 . In recent years , it has been proved that low glycosylated IgA1 is the key pathogenic factor of IgA nephropathy . The main difference between IgA1 and IgA2 is that IgA1 has a hinge region .

In this study , we studied the relationship between the activation of B - cell , IgA1 and its glycosylation in peripheral blood of patients with IgA nephropathy and the control group , and analyzed the relationship between the activation of B - cell , IgA1 and its glycosylation in peripheral blood , and provided a new theoretical basis and therapeutic target for the prevention and treatment of IgA nephropathy .

The comparison of BAFF and IgA1 in peripheral blood of patients with IgA nephropathy

Objective : To detect the expression of IgA1 in peripheral blood mononuclear cells ( PBMC ) before and after the removal of palatine tonsilla , and to analyze the relationship between the activation of the palate and the activation of B cells and IgA1 .

Methods : 16 cases of IgA nephropathy patients ( 36 - 60h ) were collected from peripheral blood of IgA nephropathy group ( 36 - 60h ) before operation ( 36 - 60h ) . 16 cases of non - IgA nephropathy group ( non - IgAN group ) and 16 normal control group ( healthy control group ) were detected by real - time quantitative PCR ( qRT - PCR ) .

Results :

1.qRT - PCR results suggest that the expression level of BAFF mRNA in IgA nephropathy patients is higher than that in normal control group ( P0.01 ) , and the expression of BAFF in IgA nephropathy patients is higher than that in normal control group ( P0.01 ) , which is higher than that in non - IgA nephropathy patients ( P0.05 ) .


2 . The content of IgA1 in plasma of IgA nephropathy patients was higher than that in normal control group ( P0.05 ) , and the content of IgA1 in plasma of IgA nephropathy patients was higher than that in normal control group ( P0.01 ) , and higher than that in non - IgA nephropathy patients ( P0.05 ) .


3 . Correlation analysis suggested that the expression of BAFF mRNA in PBMC was positively correlated with the content of IgA1 in peripheral blood ( r = 0.6604 , P0.01 ) .

Conclusion : In this chapter , the expression of BAFF gene in PBMC of patients with IgA nephropathy is increased and the expression of BAFF and its gene expression in peripheral blood plasma is closely related to BAFF . BAFF and its gene expression are expected to be a non - invasive detection index for assessing the course of IgA nephropathy in patients with IgA nephropathy .

The correlation between the expression of ST6GaINAc 鈪，

本文編號(hào)：1840364