本文選題：IgA腎病 + TLR4��； 參考：《南昌大學(xué)》2014年碩士論文

【摘要】：目的： 初步探討Toll受體4（Toll-like receptor-4,TLR4）及其調(diào)控因子細(xì)胞因子信號(hào)傳導(dǎo)抑制因子（3suppressor of cytokine signaling-3,SOCS3）在人類IgA腎病(IgAnephrology,IgAN)中的作用及機(jī)制。 方法： 收集IgAN患者及健康正常人(對(duì)照組)的一般資料及IgAN患者的臨床、病理資料。采用Ficoll法分離21例IgAN患者和21例正常人的外周血單個(gè)核細(xì)胞(peripheral blood mononuclear cell,PBMC)，實(shí)時(shí)熒光定量PCR技術(shù)（real-timepolymerase chain reaction,real-time PCR）檢測(cè)IgAN患者和正常對(duì)照組PBMC中TLR4和SOCS3mRNA的相對(duì)表達(dá)量；蛋白免疫印跡法（Western blotting,WB）檢測(cè)IgAN患者和正常人PBMC中TLR4和SOCS3的蛋白相對(duì)表達(dá)量；免疫組織化學(xué)染色（Immunohistochemistry staining,IHC）方法觀察TLR4和SOCS3在21例IgAN患者腎組織和5例腎腫瘤行腎切除患者正常腎組織的表達(dá)定位并進(jìn)行半定量分析。比較IgAN患者外周血及腎組織中TLR4和SOCS3表達(dá)量與健康對(duì)照組間的差異，相關(guān)分析法分析其與臨床、病理資料的相關(guān)性。 結(jié)果： （1）Real-time PCR結(jié)果：IgAN患者PBMC中TLR4和SOCS3mRNA的表達(dá)水平較正常對(duì)照組增高，差異無統(tǒng)計(jì)學(xué)意義。 （2）Western blotting結(jié)果：IgAN患者PBMC中TLR4和SOCS3蛋白的表達(dá)水平較正常對(duì)照組升高，其中TLR4蛋白表達(dá)量顯著高于正常對(duì)照組，差異有統(tǒng)計(jì)學(xué)意義（P 0.01）。 （3）免疫組織化學(xué)結(jié)果：TLR4和SOCS3在腎切除患者正常腎組織的腎小球、腎小管中僅有少量表達(dá)，而在IgAN患者腎組織的腎小球、腎小管中的表達(dá)量明顯高于正常對(duì)照組。 （4）相關(guān)性分析結(jié)果：IgAN患者腎組織中，TLR4在腎小球中的表達(dá)量與尿紅細(xì)胞計(jì)數(shù)成正相關(guān)，在腎小管中的表達(dá)量與血肌酐、血尿素氮、血尿酸均成正相關(guān)，與腎小球?yàn)V過率成負(fù)相關(guān)；SOCS3在腎小球中的表達(dá)量與24小時(shí)尿蛋白定量成正相關(guān)，與血漿白蛋白、總蛋白成負(fù)相關(guān)，在腎小管中的表達(dá)量與尿隱血成正相關(guān)。IgAN患者PBMC中SOCS3蛋白相對(duì)表達(dá)量與TLR4蛋白相對(duì)表達(dá)量成正相關(guān)。 結(jié)論： IgAN患者PBMC及腎組織中TLR4和SOCS3的表達(dá)量均較正常對(duì)照組升高，，且與腎臟損害程度正相關(guān)，提示TLR4和SOCS3可能均參與了IgAN的發(fā)生發(fā)展。
[Abstract]:Objective:To investigate the role and mechanism of Toll receptor 4(Toll-like receptor-4 (TLR4) and its regulatory cytokine signal transduction suppressor of cytokine signaling-3SSOCS3 in human IgA nephropathy.Methods:The general data of IgAN patients and healthy people (control group) and the clinical and pathological data of IgAN patients were collected.Peripheral blood mononuclear cells were isolated from 21 patients with IgAN and 21 normal controls by Ficoll method. The relative expressions of TLR4 and SOCS3mRNA in IgAN patients and controls were detected by real-time quantitative PCR technique.Western blotting method was used to detect the relative expression of TLR4 and SOCS3 in PBMC of IgAN patients and normal controls.Immunohistochemical staining was performed to observe the expression of TLR4 and SOCS3 in renal tissues of 21 IgAN patients and 5 cases of renal neoplasms undergoing nephrectomy and to carry out semi-quantitative analysis.The expression of TLR4 and SOCS3 in peripheral blood and kidney of patients with IgAN were compared with those in healthy controls. The correlation between the expression of TLR4 and SOCS3 in peripheral blood and renal tissues and the clinical and pathological data was analyzed by correlation analysis.Results:The results of Real-time PCR showed that the expression of TLR4 and SOCS3mRNA in PBMC of the patients with IgAN was higher than that of the control group, but the difference was not statistically significant.The results of Western blotting showed that the expression of TLR4 and SOCS3 protein in PBMC was higher than that in the control group, and the expression of TLR4 protein was significantly higher than that in the normal control group (P 0.01).(3) Immunohistochemical results showed that the expression of TLR4 and SOCS3 in the glomeruli of normal renal tissues of patients with nephrectomy was only a little in renal tubules, but the expression of TLR4 and SOCS3 in renal tubules of patients with IgAN was significantly higher than that of normal controls.The results of correlation analysis showed that the expression of TLR4 in glomeruli and renal tubules were positively correlated with serum creatinine, blood urea nitrogen and uric acid.There was a negative correlation between the expression of SOCS3 in glomeruli and the quantity of 24 hour urine protein, and a negative correlation between the expression of SOCS3 and plasma albumin and total protein.The expression of SOCS3 protein in renal tubules was positively correlated with the urine occult blood. The relative expression of SOCS3 protein was positively correlated with the relative expression of TLR4 protein in IgAN patients.Conclusion:The expression of TLR4 and SOCS3 in PBMC and renal tissues of IgAN patients was higher than that in normal controls, and was positively correlated with the degree of renal damage, suggesting that both TLR4 and SOCS3 might be involved in the pathogenesis and development of IgAN.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R692.31

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本文編號(hào)：1767368