急性肾损伤早期诊断氨基酸标志物的发现与评价

发布时间：2018-06-04 00:27

  本文选题：氨基酸 + 高效液相色谱串联质谱法　； 参考：《第二军医大学》2017年硕士论文

【摘要】：【背景及目的】急性肾损伤(Acute Kidney Injury,AKI)是肾脏结构和功能受损的一种疾病状态,以突发和持续的肾功能下降为特征,是由不同病因导致的、涉及全身各系统症状的临床综合征。2013年我国约有290万住院患者罹患AKI,死亡率约24%。AKI的发生增加了住院患者医疗费用和死亡率,同时造成医疗资源的浪费。目前AKI的诊断主要依靠血肌酐的升高和尿量的减少,而这两个指标对AKI的诊断较为滞后,不能及时反映患者肾脏的真实损伤情况,发掘新的有效AKI早期诊断标志物有助于解决当前AKI的诊断不足。肾脏参与体内多种氨基酸的代谢与排泄过程,体内氨基酸水平受到肾脏功能的调节,肾脏功能受到损伤时常发生氨基酸代谢紊乱。近年来研究发现,多种氨基酸水平与肾脏疾病患者的肾功能和预后显著相关,如非对称二甲基精氨酸、对称二甲基精氨酸、S-腺苷同型半胱氨酸等。由此可见氨基酸可能成为发掘有效AKI早期诊断标志物新的研究群体。本研究应用课题组前期建立的靶向氨基酸定量分析技术,以肾脏缺血再灌注大鼠和心脏手术患者为研究对象,进行氨基酸分析和AKI标志物筛选。AKI是急性肾病的一种状态同时与慢性肾病关系密切,一方面AKI可以在慢性肾病的基础上发生,另一方面部分AKI患者肾功能未能完全恢复也可能进展为慢性肾损伤。因此,我们还对慢性肾病患者体内氨基酸代谢紊乱情况进行了研究,探索氨基酸标志物对慢性肾病的诊断作用,以拓展氨基酸标志物的应用范围。【材料与方法】1、建立大鼠肾脏缺血再灌注模型,采集AKI发生后24小时内的多时间点血清样本进行23种氨基酸定量分析并筛选氨基酸标志物,采用受试者工作曲线评估氨基酸标志物对AKI的诊断情况,并与肌酐、尿素氮以及中性粒细胞明胶酶相关载脂蛋白进行比较。2、以心脏手术患者为研究对象,采集患者术后24小时内的多时间点血浆样本进行23种氨基酸定量分析。筛选氨基酸标志物,并对标志物进行AKI诊断分析。3、收集慢性肾病患者和健康志愿者血浆样本,筛选两组患者间差异氨基酸,分析差异氨基酸对慢性肾病的诊断作用。【研究结果】1、肾脏缺血再灌注大鼠血浆对称二甲基精氨酸、犬尿氨酸、马尿酸和氧脯氨酸水平在损伤后24小时内均对AKI具有诊断作用。四种氨基酸标志物从肾脏损伤后1小时开始对AKI具有较好的诊断作用,其中犬尿氨酸和马尿酸对AKI的诊断作用最佳,诊断能力高于中性粒细胞明胶酶相关载脂蛋白和血肌酐水平。2、心脏手术患者血浆对称二甲基精氨酸和马尿酸水平在术后多个时间具有AKI诊断作用。对称二甲基精氨酸从12小时开始对AKI具有诊断作用,且诊断能力高于术后12小时血肌酐;马尿酸水平在术后8小时开始对AKI具有诊断作用,诊断效能与12小时血肌酐水平相当,但诊断时间更早。3、对称二甲基精氨酸作为肾功能标志物不仅可以用于AKI的诊断,对慢性肾病也具有较高的诊断能力。此外,慢性肾病患者体内氨基酸代谢紊乱还包括色氨酸、酪氨酸、赖氨酸、缬氨酸、对称二甲基精氨酸、甲硫氨酸、组氨酸和亮氨酸;这些氨基酸水平均对慢性肾病具有较好的诊断效果。【结论】研究结果表明,课题组建立的靶向氨基酸技术可应用于肾前性因素导致的AKI,并由此发现具有早期诊断功能的代谢标志物,其中对称二甲基精氨酸和马尿酸水平在大鼠AKI模型中以及心脏手术患者中均具有较好的早期诊断效果。此外,临床研究结果表明对称二甲基精氨酸还能应用于慢性肾病的诊断。
[Abstract]:[background and purpose] Acute Kidney Injury (AKI) is a disease state of impaired renal structure and function. It is characterized by sudden and persistent decline in renal function. It is caused by different causes. In the year of the clinical syndrome involving systemic symptoms, about 2 million 900 thousand of our hospitalized patients suffered from AKI and the mortality rate was about 24%.AKI in.2013. At present, the diagnosis of AKI mainly depends on the increase of serum creatinine and the decrease of urine volume, and the diagnosis of the two indexes is lagging behind the diagnosis of AKI, which can not reflect the real damage of the kidney in time, and discover new effective AKI early diagnostic markers. It helps to solve the deficiency of the current AKI diagnosis. The kidney participates in the metabolism and excretion of various amino acids in the body. The level of amino acids in the body is regulated by the renal function and the renal function is damaged by the impairment of amino acid metabolism. In recent years, many amino acid levels have been found to be significantly related to renal function and prognosis in patients with renal disease. Such as asymmetric two methyl arginine, symmetric two methyl arginine, S- adenosine homocysteine, and so on. This shows that amino acids may be a new research group to explore the early diagnostic markers of effective AKI. This study used the target amino acid quantitative analysis technique established by the group in the early stage for renal ischemia reperfusion rats and patients with cardiac surgery. For the study, amino acid analysis and AKI marker screening.AKI are one of the states of acute kidney disease and chronic kidney disease. On the one hand, AKI can occur on the basis of chronic kidney disease. On the other hand, some AKI patients have not fully recovered the renal function and may progress to chronic renal injury. Therefore, we also have chronic kidney disease. The disorder of amino acid metabolism in the patient was studied in order to explore the diagnostic effect of amino acid markers on chronic kidney disease in order to expand the application of amino acid markers. [materials and methods] 1, a rat model of renal ischemia reperfusion was established, and 23 kinds of amino acid quantitation were collected from the multiple time point serum samples within 24 hours after the occurrence of AKI. Analysis and screening of amino acid markers, using the subjects' work curve to evaluate the diagnosis of amino acid markers on AKI, and compare.2 with creatinine, urea nitrogen and neutrophil gelatinase related apolipoprotein. In heart surgery patients, 23 kinds of ammonia were collected at 24 hours after the operation. Quantitative analysis of basic acid, screening of amino acid markers, and AKI diagnosis and analysis of the markers.3, collection of chronic renal disease and healthy volunteers plasma samples, screening two groups of patients with differential amino acids, analysis of differential amino acids for chronic renal disease diagnosis. [results] 1, renal ischemia reperfusion rat plasma symmetric two methyl semen The levels of ammonia, canine urinary ammonia, uric acid and proline were all diagnostic to AKI within 24 hours after injury. The four amino acid markers had a better diagnostic effect on AKI from 1 hours after renal injury. The diagnostic ability of canine urinary ammonia and uric acid for AKI was better than that of neutrophil gelatinase related load. The level of lipoprotein and serum creatinine was.2, and the plasma symmetric two methylarginine and the level of dianine had a AKI diagnostic effect at many times after the operation. Symmetric two methyl arginine had a diagnostic effect on AKI from 12 hours, and the diagnostic ability was higher than 12 hours after the operation; the level of urine acid began to AKI at 8 hours after the operation. The diagnostic efficacy is equal to 12 hours serum creatinine level, but the diagnosis time is earlier.3. Symmetric two methyl arginine as a renal function marker can not only be used in the diagnosis of AKI, but also have high diagnostic ability for chronic kidney disease. Valine, symmetrical two methyl arginine, methionine, histidine, and leucine, these amino acid levels have a good diagnostic effect on chronic kidney disease. [Conclusion] the results of the study suggest that the target amino acid technique established by the group can be applied to AKI induced by pre renal factors, and thus the metabolic markers that have early diagnostic functions are found. In the AKI model of rats and the patients with cardiac surgery, symmetrical two methyl arginine and dianine have good early diagnosis effect. In addition, the clinical results show that symmetric two methylarginine can be used in the diagnosis of chronic kidney disease.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R692

【参考文献】

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