TLR4及其调控因子SOCS3在IgA肾病发病机制中作用的初步探讨

发布时间：2018-04-18 07:27

本文选题：IgA肾病 + TLR4　；参考：《南昌大学》2014年硕士论文

【摘要】：目的：初步探讨Toll受体4（Toll-like receptor-4,TLR4）及其调控因子细胞因子信号传导抑制因子（3suppressor of cytokine signaling-3,SOCS3）在人类IgA肾病(IgAnephrology,IgAN)中的作用及机制。方法：收集IgAN患者及健康正常人(对照组)的一般资料及IgAN患者的临床、病理资料。采用Ficoll法分离21例IgAN患者和21例正常人的外周血单个核细胞(peripheral blood mononuclear cell,PBMC)，实时荧光定量PCR技术（real-timepolymerase chain reaction,real-time PCR）检测IgAN患者和正常对照组PBMC中TLR4和SOCS3mRNA的相对表达量；蛋白免疫印迹法（Western blotting,WB）检测IgAN患者和正常人PBMC中TLR4和SOCS3的蛋白相对表达量；免疫组织化学染色（Immunohistochemistry staining,IHC）方法观察TLR4和SOCS3在21例IgAN患者肾组织和5例肾肿瘤行肾切除患者正常肾组织的表达定位并进行半定量分析。比较IgAN患者外周血及肾组织中TLR4和SOCS3表达量与健康对照组间的差异，相关分析法分析其与临床、病理资料的相关性。结果：（1）Real-time PCR结果：IgAN患者PBMC中TLR4和SOCS3mRNA的表达水平较正常对照组增高，差异无统计学意义。（2）Western blotting结果：IgAN患者PBMC中TLR4和SOCS3蛋白的表达水平较正常对照组升高，其中TLR4蛋白表达量显著高于正常对照组，差异有统计学意义（P 0.01）。（3）免疫组织化学结果：TLR4和SOCS3在肾切除患者正常肾组织的肾小球、肾小管中仅有少量表达，而在IgAN患者肾组织的肾小球、肾小管中的表达量明显高于正常对照组。（4）相关性分析结果：IgAN患者肾组织中，TLR4在肾小球中的表达量与尿红细胞计数成正相关，在肾小管中的表达量与血肌酐、血尿素氮、血尿酸均成正相关，与肾小球滤过率成负相关；SOCS3在肾小球中的表达量与24小时尿蛋白定量成正相关，与血浆白蛋白、总蛋白成负相关，在肾小管中的表达量与尿隐血成正相关。IgAN患者PBMC中SOCS3蛋白相对表达量与TLR4蛋白相对表达量成正相关。结论： IgAN患者PBMC及肾组织中TLR4和SOCS3的表达量均较正常对照组升高，，且与肾脏损害程度正相关，提示TLR4和SOCS3可能均参与了IgAN的发生发展。
[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.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R692.31

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