【摘要】：目的:應(yīng)用基因測序的方法分析特發(fā)性膜性腎病患者血清及尿液外泌體中環(huán)狀RNA的差異表達(dá),為進(jìn)一步探索環(huán)狀RNA作為特發(fā)性膜性腎病診斷生物標(biāo)志物及其調(diào)控機(jī)制研究奠定基礎(chǔ)。方法:特發(fā)性膜性腎病患者(IMN組)及正常健康對照者(NC組)各取10例為實驗對象,分別收集其外周靜脈血分離出的血清及晨尿,并應(yīng)用試劑盒及超速離心法提取各組的外泌體,一系列酶解反應(yīng)獲取外泌體中的環(huán)狀RNA,然后利用基因測序的方法篩選出差異性表達(dá)的環(huán)狀RNA,并對其靶基因進(jìn)行分析。結(jié)果:與健康對照組相比,特發(fā)性膜性腎病患者血清外泌體中環(huán)狀RNA的種類減少,且主要表現(xiàn)為內(nèi)含子來源,顯著差異性表達(dá)的有89種,主要也是內(nèi)含子來源,其中49種表達(dá)上調(diào),40種表達(dá)下調(diào);而尿液外泌體中環(huán)狀RNA的種類增多,且主要表現(xiàn)為外顯子來源,顯著差異性表達(dá)的有60種,但主要是內(nèi)含子來源,其中54種表達(dá)上調(diào),6種表達(dá)下調(diào)。膜性腎病患者血清與尿液外泌體相比,顯著差異性表達(dá)的環(huán)狀RNA有59種,且主要是內(nèi)含子來源,其中32種表達(dá)上調(diào),27種表達(dá)下調(diào)。結(jié)論:特發(fā)性膜性腎病患者血清及尿液外泌體中環(huán)狀RNA的表達(dá)具有疾病特異性。顯著差異和特異性表達(dá)的環(huán)狀RNA,如chr7:100550808|100551062對應(yīng)的MUC3A基因可作為特發(fā)性膜性腎病潛在的生物學(xué)標(biāo)志物,并且可為特發(fā)性膜性腎病發(fā)病機(jī)制的研究提供參考和補(bǔ)充。
文內(nèi)圖片：
圖片說明：血清外泌體沉淀圖
[Abstract]:Objective: to analyze the differential expression of annular RNA in serum and urine exocrine bodies of patients with idiopathic membranous nephropathy by gene sequencing, and to lay a foundation for further exploring the use of annular RNA as a biomarker for the diagnosis of idiopathic membranous nephropathy and its regulatory mechanism. Methods: 10 patients with idiopathic membranous kidney disease (IMN group) and 10 normal controls (NC group) were selected as experimental subjects. The serum and morning urine isolated from peripheral venous blood were collected, and the exocrine bodies of each group were extracted by kit and ultracentrifugation. A series of enzyme hydrolysis reactions were used to obtain annular RNA, in exocrine. Then the differentially expressed circular RNA, was screened by gene sequencing and its target gene was analyzed. Results: compared with the healthy control group, the type of cyclic RNA in serum exocrine of patients with idiopathic membranous nephropathy decreased, and the main expression was intron source. 89 kinds of significantly differentially expressed, mainly intron source, of which 49 kinds of expression were up-regulated and 40 kinds of expression were down-regulated. However, the types of cyclic RNA in urine exocrine increased, and the main expression was exons. There were 60 kinds of significantly differentially expressed, but mainly intron sources, of which 54 were up-regulated and 6 were down-regulated. Compared with urine exocrine, there were 59 kinds of annular RNA differentially expressed in serum and urine of patients with membranous nephropathy, and they were mainly derived from introns, of which 32 were up-regulated and 27 were down-regulated. Conclusion: the expression of cyclic RNA in serum and urine exocrine bodies of patients with idiopathic membranous nephropathy is disease-specific. The MUC3A gene corresponding to significant difference and specific expression of circular RNA, such as chr7:100550808 / 100551062, can be used as a potential biomarker for idiopathic membranous nephropathy, and can be used as a reference and supplement for the study of the pathogenesis of idiopathic membranous nephropathy.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R692

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