Pentraxin 3与糖尿病肾脏病的相关性研究

发布时间：2018-01-15 07:16

本文关键词：Pentraxin 3与糖尿病肾脏病的相关性研究　出处：《南京医科大学》2017年博士论文　论文类型：学位论文

【摘要】：研究背景和目的:糖尿病肾脏病(DKD)是糖尿病常见的微血管并发症,遗传和环境因素在DKD发生发展中起着重要的作用。Pentraxin 3(PTX3)是长链PTX的一员,与短链PTX,如C反应蛋白(CRP)和血清淀粉样蛋白P(SAP)在结构上同源,但是不同于CRP主要是在肝脏合成,PTX3是在炎症信号刺激下,主要在外周组织和细胞分泌表达。在肾组织中,PTX3是唯一被检测到表达的长链PTX。PTX3是急性期糖蛋白,其作为调理素的可溶性受体,在体液先天免疫中以及在变性和凋亡的细胞清除中起作用;而且PTX3结合多种配体,激活补体系统等。PTX3基因位于染色体3q25,全基因组连锁分析显示染色体3q与糖尿病肾脏病关联。既往临床研究显示血浆PTX3浓度与糖尿病肾脏病相关。本课题首次报道PTX3基因多态性与DKD易感性的相关性研究,另在早期DKD大鼠肾组织中初步分析PTX3的表达以及和肾损伤的关系。旨在探讨PTX3与DKD的关系,为早期发现或防治DKD探索新的可能指标或治疗靶点。研究方法:1、采用病例-对照的方法在135例2型糖尿病肾脏病患者(DKD)、155例没有并发DKD的2型糖尿病患者(NDKD)和152例正常对照者(NC)中,应用多重 SNaPshot SNPs 分型技术对 PTX3 基因的 8 个位点(rs2305619,rs2120243,rs1456099,rs7634847,rs1840680,rs2316706,rs2316709 和 rs7616177)进行分型。2、单次腹腔注射STZ建立糖尿病大鼠模型,12周后检测尿液白蛋白肌酐比和总蛋白肌酐比,判定早期DKD模型建立成功。早期DKD模型大鼠随机分成2组,胰岛移植治疗组(IT组)和无治疗的DKD组(DKD组)。并于16周麻醉后处死大鼠收集标本。免疫组织化学染色和western blot检测大鼠肾组织中PTX3及其他相关肾脏损伤指标(Bax、Bcl-2、Caspase-3、NPHS-2、alpha-SMA 和 TGF-beta)的表达情况。结果:1、PTX3基因位点rs2305619和rs2120243的基因型分布频率在DKD组和NDKD组间差异有统计学意义(分别P=0.017,P=0.033)。2型糖尿病患者中携带rs2305619 GG基因型的罹患DKD的风险是AA基因型的4.078倍(95%CI=1.370-12.135,P =0.012)。携带 rs2120243 AA 基因型的 2 型糖尿病患者与CC基因型相比患DKD的风险下降(OR=0.213,95%CI=0.055-0.826,P=0.025)。2、单倍型 CAGGG(SNPs 位点:rs2120243-rs2316709-rs7616177-rs2305619-rs1840680)与罹患DKD的风险增高有关(P=0.0319)。3、PTX3的表达位点主要在肾小管上皮细胞,早期DKD大鼠模型肾组织中,肾小管上皮细胞大量凋亡,肾小管PTX3的表达明显减少(P0.01)。DKD大鼠胰岛移植后随着肾脏细胞凋亡及纤维化等损伤指标改善,DKD得到了逆转。同时发现肾小管PTX3表达比DKD组明显升高(P0.01),接近于正常大鼠组。结论:1、PTX3基因SNPs与2型糖尿病肾脏病易感性相关,PTX3基因可能是中国汉族2型糖尿病患者中罹患DKD的候选易感基因。2、PTX3在DKD大鼠肾组织中的表达与肾损伤密切相关:在早期糖尿病肾脏病大鼠肾小管上皮细胞中表达下降。胰岛移植逆转DKD后,PTX3表达明显升高。
[Abstract]:Background and objective: diabetic kidney disease (DKD) is a common microvascular complication of diabetes mellitus. Genetic and environmental factors play an important role in the occurrence and development of DKD. Pentraxin 3 (PTX3) is a member of long chain PTX and short strand PTX. For example, C-reactive protein (CRP) and serum amyloid protein (PS-SAP) are structurally homologous, but unlike CRP, they are mainly synthesized in the liver and stimulated by inflammatory signals. PTX3 is the only long chain PTX.PTX3 detected in renal tissue, which is a glycoprotein in acute phase and is a soluble receptor for modulin. Play a role in innate humoral immunity and in the removal of degenerative and apoptotic cells; Moreover, PTX3 binds multiple ligands and activates complement system. PTX3 gene is located on chromosome 3q25. Genomic linkage analysis showed that chromosome 3q was associated with diabetic kidney disease. Previous clinical studies showed that plasma PTX3 concentration was associated with diabetic kidney disease. This study reported for the first time that PTX3 gene polymorphism and DK were associated with diabetic kidney disease. Correlation study of D susceptibility. In addition, the expression of PTX3 and the relationship between PTX3 expression and renal injury were analyzed in the early stage of DKD rats. The purpose of this study was to explore the relationship between PTX3 and DKD. In order to find or prevent and treat DKD in early stage, a new possible index or therapeutic target was explored. Method: 1. A case-control method was used in 135 patients with type 2 diabetic kidney disease (type 2 diabetic kidney disease). There were 155 patients with type 2 diabetes mellitus without DKD and 152 normal controls. Multiple SNaPshot SNPs typing technique was used to detect 8 PTX3 loci, rs2305619 / rs2120243rs1456099. Rs7634847, rs1840680, rs2316706, rs2316709 and rs7616177). The urine albumin creatinine ratio and total protein creatinine ratio were measured 12 weeks after the diabetic model was established by single intraperitoneal injection of STZ. Early DKD model rats were randomly divided into two groups. Islet transplantation therapy group (IT group) and untreated DKD group (DKD group). After 16 weeks of anesthesia, the rats were killed to collect the specimens, and the PTX3 and other related renal damage indexes were detected by immunohistochemical staining and western blot. The expression of alpha-SMA and TGF-beta in Bcl-2Caspase-3, NPHS-2 and TGF-beta. The genotypic frequencies of rs2305619 and rs2120243 in PTX3 gene locus were significantly different between DKD group and NDKD group (P0. 017, respectively). The risk of developing DKD in patients with rs2305619 GG genotype was 4.078 times higher than that in AA patients (P < 0.05). 95CII 1.370-12.135. The risk of DKD in type 2 diabetic patients with rs2120243 AA genotype was lower than that with CC genotype, and the risk of type 2 diabetes mellitus patients with rs2120243 AA genotype was lower than that of CC genotype. 95. CIQ 0.055-0.826 P0. 025. 2. Haplotype CAGGG(SNPs locus: rs2120243-rs2316709-rs7616177-rs2305619-rs1840680). It was associated with increased risk of DKD. The expression site of PTX3 was mainly located in renal tubular epithelial cells, early DKD rat model kidney tissue, renal tubular epithelial cells apoptosis. The expression of PTX3 in renal tubules decreased significantly with the improvement of renal cell apoptosis and fibrosis after islet transplantation. DKD was reversed. It was also found that the expression of PTX3 in renal tubules was significantly higher than that in DKD group, which was close to that in normal rats. Conclusion 1. PTX3 gene SNPs is associated with susceptibility to type 2 diabetic kidney disease. PTX3 gene may be a candidate susceptibility gene for DKD in type 2 diabetic patients of Han nationality in China. The expression of PTX3 in renal tissue of DKD rats was closely related to renal injury: the expression of PTX3 was decreased in renal tubular epithelial cells of early diabetic nephropathy rats, and DKD was reversed by islet transplantation. The expression of PTX3 was significantly increased.
【学位授予单位】：南京医科大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R587.2;R692.9

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