【摘要】：研究背景：膜性腎病(MN, membranous nephropathy)和微小病變性腎病(MCN, minimal change nephropathy)分別是成人和兒童腎病綜合征中最常見的病理類型,它們臨床上都是以大量蛋白尿、低白蛋白血癥、水腫和高脂血癥為主要特點(diǎn),病理改變上兩者都會(huì)發(fā)生足突細(xì)胞的融合,同時(shí)也有各自特征性的改變。長期大量蛋白尿?qū)δI病患者的預(yù)后具有極大的影響,近年來,以MCD大鼠為研究對(duì)象的研究發(fā)現(xiàn)：與腎病綜合征其他病理類型相比, MCD大鼠腎組織中血管生成素樣蛋白-4(Angpt14, Angiopoietin-like4)的表達(dá)明顯升高。 目的：通過對(duì)比分析足細(xì)胞損傷腎病綜合征患者、足細(xì)胞損傷非腎病綜合征患者、IgA腎病患者和正常健康對(duì)照者的血漿、尿液和腎組織石蠟切片中Angpt14的表達(dá)特點(diǎn)及差異。 實(shí)驗(yàn)方法：實(shí)驗(yàn)分四組,分別為足細(xì)胞損傷腎病綜合征組(NS組,nephrotic syndrome group)72例、足細(xì)胞損傷非腎病綜合征組(non-NS組)64例、IgA腎病組(IgAN組,IgA nephropathy group)37例和健康對(duì)照組60例(Con組,healthy control group),收集四組個(gè)體的血漿、尿液和腎組織石蠟切片標(biāo)本,運(yùn)用酶聯(lián)免疫吸附劑測(cè)定法(ELISA, enzyme linked immunosorbent assay)測(cè)定Angpt14蛋白的表達(dá)情況,并用方差分析(Analysis of Variance)進(jìn)行數(shù)據(jù)分析。應(yīng)用免疫組化測(cè)定腎活檢組織石蠟切片中Angptl4蛋白的表達(dá)情況。 結(jié)果：血漿中,NS組、non-NS組、IgAN組和Control組之間Angptl4OD值比較,結(jié)果除NS組與non-NS組之間沒有差異外,其余兩兩之間均有顯著差異(NS:1.296±0.634; non-NS:1.469±0.546; IgAN:1.731±0.387; Control:2.142±0.416),比較IgAN組中尿蛋白肌酐比值不同的三組患者(0-0.5g/g vs0.5-3.5g/g vs3.5-g/g)的血漿Angpt140D值發(fā)現(xiàn),三者之間有差異(1.875±0.381vs1.694±0.324vs1.016±0.098).同樣對(duì)non-NS組中尿蛋白肌酐比值不同的兩組患者和NS組進(jìn)行了比較(0-0.5g/g vs0.5-3.5g/g vs NS組),三者之間沒有顯著差異。應(yīng)用線性相關(guān)分析發(fā)現(xiàn)NS組中血漿Angpt14OD值與患者尿蛋白肌酐比值、尿蛋白濃度、GFR以及Scr之間均沒有顯著相關(guān)關(guān)系。IgAN組中血漿Angpt14與尿蛋白肌酐和尿蛋白濃度有線性相關(guān)關(guān)系。分析non-NS組,發(fā)現(xiàn)血漿Angpt14OD值與患者尿蛋白濃度之間呈負(fù)相關(guān)關(guān)系。 尿液中,NS組、non-NS組、IgAN組和Control組之間Angpt14OD值比較顯示兩兩之間均有顯著差異(NS:2.830±0.340; non-NS組：2.456±0.770; IgAN:1.569±0.810; Control:0.163±0.270)。比較IgA組中尿蛋白肌酐比值不同的三組患者(0-0.5g/g vs0.5-3.5g/g vs3.5-g/g)的尿液Angptl4OD值發(fā)現(xiàn),兩者之間沒有顯著差異。同樣對(duì)non-NS組中尿蛋白肌酐比值不同的兩組患者與NS組進(jìn)行了比較(0-0.5g/g vs0.5-3.5g/g vs NS組),發(fā)現(xiàn)兩兩之間均有顯著差異(1.150±0.086vs2.537±0.717vs2.830±0.340).線性相關(guān)分析發(fā)現(xiàn)NS組中尿液Angpt14OD值與患者尿蛋白肌酐比值、尿蛋白濃度、GFR以及Scr之間均沒有顯著相關(guān)關(guān)系。IgAN組中發(fā)現(xiàn)尿液Angpt14的OD值與尿蛋白肌酐比值以及尿蛋白濃度均呈正相關(guān)。分析non-NS組,發(fā)現(xiàn)尿液Angpt14OD值與患者尿蛋白肌酐比值之間也呈正相關(guān)關(guān)系。 免疫組化結(jié)果顯示,NS組和non-NS組的足細(xì)胞均有陽性染色,IgAN組和Control組腎小球足細(xì)胞均為陰性,其中NS組的陽性染色高于non-NS組。 結(jié)論：足細(xì)胞損傷腎病綜合征患者、足細(xì)胞損傷非腎病綜合征患者、IgA腎病患者和正常健康對(duì)照者(兩兩之間)的血漿、尿液和腎組織Angptl4的表達(dá)有差異。Angpt14在足細(xì)胞損傷患者,尤其是腎病綜合征患者中有特異性表達(dá),尿液中Angpt14蛋白的差異可以反映足細(xì)胞損傷程度的差異。該蛋白在足細(xì)胞損傷的病理生理過程中有重要作用。
[Abstract]:Background: The most common pathological types in adult and children's nephrotic syndrome are membranous nephropathy (MN, mbinous nephron) and micro-pathological renal disease (MCN, minimbal change nephron), which are mainly characterized by a large amount of proteinuria, hypoalbuminemia, edema and hyperlipidemia. The fusion of the apophysis cells can occur both in the pathological change and the characteristic changes. In recent years, the expression of angiopoietin-like protein-4 (Angpt14, Angiopoietin-like4) in the kidney of the MCD was significantly increased compared with the other pathological types of the nephrotic syndrome. Objective: To compare the expression and difference of Angpt14 in serum, urine and renal tissue paraffin sections of patients with nephrotic syndrome, diabetic nephropathy and normal healthy controls. Methods: The experimental methods were divided into four groups: group NS group, nephrastic syndrome group (72 cases), foot cell injury non-nephrotic syndrome group (non-NS group)64 cases, IgA nephropathy group (IgAN group, IgA nephron group)37 cases and healthy control group 60 cases (Con group, healthy control gro). Up), blood plasma, urine and kidney tissue paraffin section specimens of four groups were collected, and the expression of the Angpt14 protein was determined by an enzyme-linked immunosorbent assay (ELISA, enzyme linked immunosorbent assay) and the number of analysis of variance was used to count the expression of the Angpt14 protein. An immunohistochemical method for the determination of Angptl4 protein in paraffin section of renal biopsy tissue As a result, the Angptl4OD values between the NS group, the non-NS group, the IgAN group and the Control group were compared, and there was no difference between the NS group and the non-NS group, and there was a significant difference between the other two groups (NS: 1.296-0.634; non-NS: 1.469-0.546; IgAN: 1.731-0.387; Control: 2.142-0 .416) The plasma Angpt140D values of three groups of patients (0-0.5 g/ g vs0.5-3.5 g/ g vs3.5-g/ g) in the IgAN group were found to be different (1.875) 0.381 vs1.694 and 0.324 vs1.016-0 in comparison with the plasma Angpt140D values of the different groups of patients (0-0.5 g/ g vs0.5-3.5 g/ g vs3.5-g/ g) in the IgAN group. .0-0.5 g/ g vs0.5-3.5 g/ g vs. NS group) were also compared in two groups of patients and NS groups with different urinary protein levels in the non-NS group (0-0.5 g/ g vs0.5-3.5 g/ g vs. NS group). There was a significant difference. There was no significant difference in the ratio of Angpt14OD in the NS group to the ratio of the patient's urinary protein, the concentration of urinary protein, the GFR, and the Scr in the NS group. The relationship between plasma Angpt14 and urinary protein and urinary protein concentration in the IgAN group The relationship between plasma Angpt14OD and urinary protein concentration was found in non-NS group. There were significant differences in the Angpt14OD between the NS group, the non-NS group, the IgAN group and the Control group (NS: 2.830-0.340; non-NS group: 2.456-0.770; IgAN: 1.569-0.810; Control: 0.163 (0.270) The urine Angptl4OD values of three groups of patients (0-0.5 g/ g vs0.5-3.5 g/ g vs3.5-g/ g) in the IgA group were found to be different from those in the IgA group, both of which There was no significant difference between the two groups (0-0.5 g/ g vs0.5-3.5 g/ g vs. NS group) in the non-NS group, and there was a significant difference between the two groups (1.150, 0.086 vs2.537) 0.717 vs2.830 (0.340). The linear correlation analysis found that the urine Angpt14OD value in the NS group was not related to the ratio of the urinary protein of the patient, the concentration of the urinary protein, the GFR, and the Scr. There was a significant correlation. In the IgAN group, the ratio of OD value of the urine Angpt14 to the urinary protein muscle and the urinary protein were found in the IgAN group. The concentrations were positively correlated. The non-NS group was analyzed to find the ratio of Angpt14OD of urine to the ratio of urinary protein in the patient. The positive staining was positive in both the NS group and the non-NS group. The positive staining of the NS group was negative. Higher than non-NS group. Conclusion: The plasma, urine and renal tissue An in the patients with diabetic nephrotic syndrome, patients with foot cell injury, non-nephrotic syndrome, patients with IgA nephropathy and normal healthy controls (between two) The expression of gtl4 is different. Angpt14 is specifically expressed in patients with foot cell injury, especially in nephrotic syndrome, and the difference of Angpt14 protein in urine can be reversed. The difference of the degree of damage to the foot cell. The protein is in the disease of the foot cell.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R692

【共引文獻(xiàn)】

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