糖尿病肾病患者MIOX的表达及其临床意义研究

发布时间：2018-05-08 08:28

本文选题：糖尿病肾病 + MIOX　；参考：《中南大学》2014年硕士论文

【摘要】：研究背景糖尿病肾病(diabetic nephropathy, DN)是糖尿病微血管并发症的重要类型,也是引起终末期肾脏疾病(end stage renal disease, ESRD)的主要病因(约占40%-50%)。糖尿病肾病发病机制复杂,已有大量研究证实多元醇通路活性增高、糖基化终末产物形成增加、蛋白激酶C(protein kinase C,PKC)和RAS系统激活、GTP酶蛋白的活性变化、TGF-β-Smad信号通路的激活等细胞分子信号转导途径异常均参与了糖尿病肾病的发生与进展,但其具体的发病机制尚不完全清楚。目前临床常用针对上述单个因素及信号转导分子的治疗干预措施如PKC抑制剂等疗效有限。因此深入开展糖尿病肾病分子发病机制的研究,寻找糖尿病肾病新的治疗靶点,一直是国内外肾脏病学界研究领域的热点,具有重要的科学和社会意义。传统观念认为肾小球细胞损伤是糖尿病肾病的主要特征,但新近研究证实肾小管间质损伤在糖尿病肾病中具有重要的意义,且肾小管间质病变并不继发于肾小球病变,而是糖尿病肾病的早期和原始特征。肾小管上皮细胞损伤可导致肾小管重吸收和分泌功能异常,促进肾小管上皮细胞萎缩及肾间质纤维化。最近研究证实肌醇耗竭在糖尿病肾病肾小管间质损伤的过程中起着重要的作用,而肾脏特异性表达的肌醇加氧酶(Myo-inositol oxygenase, MIOX)主要表达在肾组织肾小管上皮细胞内,是组织中肌醇代谢的关键调节酶。体外实验表明高葡萄刺激可上调肾小管上皮细胞内肌醇加氧酶的表达,并产生促进活性氧(reactive oxygen species, ROS)生成及纤维连接蛋白(fibronectin, FN)表达的效应。基于以上分析,我们认为MIOX在糖尿病肾病患者肾组织内表达上调,并可能与糖尿病肾病肾小管上皮细胞氧化损伤及间质纤维化密切相关。故本试验中我们通过检测MIOX在糖尿病肾病患者肾活检组织及血液、尿液中的表达,并分析其余临床指标之间的相关性,旨在为进一步研究糖尿病肾病的发病机制及治疗提供新的思路与靶点。第一章糖尿病肾病患者肾组织中MIOX的表达及其与临床指标相关性分析目的：观察MIOx在糖尿病肾病患者肾组织中的表达,并分析其与临床各指标之间的关系。方法：随机选取经临床和肾脏病理活检确诊为糖尿病肾病的患者15人及轻微病变的患者(原发性肾病)15人。采用HE, PASM, Masson,PAS染色观察肾脏病理改变,免疫组化、免疫荧光检测MIOX表达病变,并使用SPSS17.0分析糖尿病肾病患者肾组织内MIOX相对表达程度与临床指标间的相关性。结果：肾脏病理显示糖尿病肾病患者肾活检组织肾小球体积增大,部分出现结节性肾小球硬化,出现典型的Kimmelstiel-Wilson结节,肾小球系膜基质的增生,基底膜的增厚、僵硬；肾小管片状萎缩并部分代偿性扩张,肾间质的片状纤维化改变。而对照组为轻微病变患者,肾小球部分基本正常,系膜细胞及系膜基质都无明显增生,肾小管间质无明显纤维化,血管无明显异常。糖尿病肾病患者的肾小管间质损伤评分明显比对照组高。免疫组化实验结果提示：MIOX主要表达在肾小管上皮细胞胞浆内。免疫荧光试验结果提示：MIOX主要表达在肾小管上皮细胞的胞浆及肾间质内。与非糖尿病肾病轻微病变相比,糖尿病肾病患者肾小管细胞内MIOX表达显著上调(P0.05)。相关性分析表明MIOX表达强度与糖尿病肾病患者肾小管间质损伤评分、血清肌酐、血糖、糖化血红蛋白水平呈正相关。结论：糖尿病肾病患者肾组织MIOX表达水平明显上升,MIOX可能参与糖尿病肾病肾小管间质损伤。第二章糖尿病肾病患者血液及尿液中MIOX的表达及其与临床指标的相关性分析目的：观察糖尿病肾病患者血清及尿液中MIOX的表达,分析其与临床指标的关系。方法：随机选取临床诊断为糖尿病肾病患者30人,非糖尿病肾病慢性肾脏疾病患者30人,单纯糖尿病患者30人,正常对照组30人。采用ELISA法检测外周血及尿液中间的MIOX表达。结果：MIOX检测结果显示：糖尿病肾病组,非糖尿病肾病组,糖尿病组,及正常对照组血清MIOX的水平分别为:2050.4士160.7、1069.2±96.23、619.3±64.1、49.47±3.3pg/ml。糖尿病肾病组,非糖尿病肾病组,糖尿病组,及正常对照组尿液MIOX的水平分别为：2675.3±241.1、1427.2±146.3、775.0±80.3、73.5±4.8pg/ml。糖尿病肾病患者血,尿MIOX水平明显高于非糖尿病肾病组、单纯糖尿病患者组及正常对照组,差异具有统计学差异(p0.01),相关性分析发现糖尿病肾病患者血清及尿液MIOX表达水平与血清肌酐、血糖、糖化血红蛋白水平呈正相关。结论：糖尿病肾病患者外周血及尿液中MIOX的表达水平明显上调。
[Abstract]:Research background
Diabetic nephropathy (DN) is an important type of diabetic microvascular complication and is also the main cause of end stage renal disease (ESRD). The pathogenesis of diabetic nephropathy is about 40%-50%. The pathogenesis of diabetic nephropathy is complex. A large number of studies have confirmed that the activity of polyol pathway is increased and the end products of glycosylation are increased. In addition, the activation of protein kinase C (protein kinase C, PKC) and RAS system, the changes in the activity of GTP enzyme protein and the activation of the TGF- beta -Smad signaling pathway are all involved in the development and progression of diabetic nephropathy, but the specific pathogenesis is not completely clear. The therapeutic intervention of number transducers, such as PKC inhibitors, is limited. Therefore, it is of great scientific and social significance to develop the molecular pathogenesis of diabetic nephropathy and find new therapeutic targets for diabetic nephropathy, which has always been a hot spot in the field of Nephrology at home and abroad.
Traditionally, glomerular cell damage is the main feature of diabetic nephropathy. However, recent studies have confirmed that renal tubulointerstitial damage is of great significance in diabetic nephropathy. Renal tubulointerstitial lesions are not secondary to glomerular lesions, but are early and original characteristics of diabetic nephropathy. Renal tubular epithelial cell damage can lead to kidney disease. The renal tubular epithelial cell atrophy and renal interstitial fibrosis are promoted by the abnormal reabsorption and secretory function of the tubules. Recent studies have shown that inositol depletion plays an important role in the process of renal tubulointerstitial damage in diabetic nephropathy, and the renal specific expression of inositol oxygenase (Myo-inositol oxygenase, MIOX) is mainly expressed in renal tissue. In vitro, it is the key regulator of the metabolism of inositol in the tissue. In vitro experiments show that high grape stimulation can increase the expression of inositol oxygenase in renal tubular epithelial cells and produce the effect of reactive oxygen species (ROS) generation and fibronectin (fibronectin, FN) expression. Based on the above analysis, we believe that M The expression of IOX is up-regulated in the renal tissue of patients with diabetic nephropathy and may be closely related to the oxidative damage and interstitial fibrosis of renal tubular epithelial cells in diabetic nephropathy. In this study, we detected the expression of MIOX in renal biopsy tissue, blood and urine in patients with diabetic nephropathy, and analyzed the correlation between the other clinical indicators. It will provide new ideas and targets for further study on the pathogenesis and treatment of diabetic nephropathy.
Chapter 1 expression of MIOX in renal tissue of patients with diabetic nephropathy and its correlation with clinical indicators
Objective: To observe the expression of MIOx in renal tissue of patients with diabetic nephropathy and analyze its relationship with clinical indicators.
Methods: 15 patients with diabetic nephropathy and 15 patients with mild pathological changes (primary nephrosis) were randomly selected by clinical and renal biopsy. HE, PASM, Masson, PAS staining were used to observe renal pathological changes, immunohistochemistry, immunofluorescence detection of MIOX expression, and SPSS17.0 analysis of MI in renal tissue of diabetic nephropathy patients. The correlation between the relative expression level of OX and clinical indicators.
Results: renal pathology showed that the glomerular volume increased in renal biopsy tissue, nodular glomerulosclerosis, typical Kimmelstiel-Wilson nodules, glomerular mesangial matrix hyperplasia, basilar membrane thickening, stiffness, renal tubuloatrophy and partial compensatory dilation, and renal interstitial fibrosis. In the control group, the glomerular part was basically normal, the mesangial cells and mesangial matrix had no obvious hyperplasia, there was no obvious fibrosis in the tubulointerstitium and no obvious abnormal blood vessels. The renal tubulointerstitial damage score of diabetic nephropathy patients was significantly higher than that of the control group. The immunohistochemical results suggested that MIOX was mainly expressed in the kidneys. The results of immunofluorescence test showed that MIOX was mainly expressed in the cytoplasm and renal interstitium of renal tubular epithelial cells. Compared with non diabetic nephropathy, the expression of MIOX in renal tubule cells in diabetic nephropathy patients was significantly up-regulated (P0.05). The correlation analysis showed that the expression intensity of MIOX was smaller than that of diabetic nephropathy. There was a positive correlation between tubulointerstitial injury score, serum creatinine, blood sugar and glycosylated hemoglobin level.
Conclusion: the expression level of MIOX in renal tissue of patients with diabetic nephropathy increases significantly, and MIOX may be involved in tubulointerstitial injury in diabetic nephropathy.
The second chapter is about the expression of MIOX in blood and urine of patients with diabetic nephropathy and its correlation with clinical indicators.
Objective: To observe the expression of MIOX in serum and urine of patients with diabetic nephropathy and analyze its relationship with clinical indicators.
Methods: 30 patients with diabetic nephropathy, 30 patients with chronic renal disease in non diabetic nephropathy, 30 patients with simple diabetes and 30 normal controls were randomly selected. The MIOX expression in the middle of peripheral blood and urine was detected by ELISA.
Results: the results of MIOX detection showed that the levels of serum MIOX in diabetic nephropathy group, non diabetic nephropathy group, diabetes group and normal control group were 2050.4 160.71069.2 + 96.23619.3 + 64.1,49.47 + 3.3pg/ml. diabetic nephropathy group, non diabetic nephropathy group, diabetes group, and normal control group urine MIOX level were 2675, respectively: The serum and urine MIOX levels of.3 + 241.11427.2 + 146.3775.0 + 80.3,73.5 + 4.8pg/ml. diabetic nephropathy patients were significantly higher than those of non diabetic nephropathy group. The difference was statistically significant (P0.01). The correlation analysis found the serum and urine MIOX expression level and serum creatinine, blood glucose, and blood glucose in patients with diabetic nephropathy. The level of glycosylated hemoglobin was positively correlated.
Conclusion: the expression level of MIOX in peripheral blood and urine of diabetic nephropathy patients is significantly increased.

【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R587.2;R692.9

【共引文献】