顺铂诱导的Beagle犬急性肾损伤模型中尿液生物标志物研究

目的:应用顺铂诱导Beagle犬急性肾损伤模型,评价新型尿液生物标志物的诊断效能.方法:通过对Beagle犬单次给予3 mg·kg-1顺铂静脉注射(iv),建立了急性肾损伤模型,应用ELISA和Luminex液相芯片方法检测分析尿液中9种新型生物标志物浓度,并与传统血清指标尿素氮(BUN)和肌酐(Cr)进行比较,结合肾...     

肾损伤患者尿液酶谱分析

将我院不同病因及不同程度的肾损伤患者58例,检测具有代表意义的7种酶进行对比分析,现报道如下:1 材料和方法1.1 样品收集:健康成人176例为对照组,年龄18-64岁,男106例,女70例。肾损伤为本院住院患者58例为观察组,经检验、B超、肾图或肾造影确诊,其中男28例,女30例,年龄15-     

急性肾损伤新的标志物NGAL的临床检测意义

急性肾损伤(AKI)以往的诊断标准通常使用血清肌酐浓度和尿量两个指标,其诊断的敏感度不高。中性粒细胞明胶酶相关脂质运载蛋白(NGAL)作为急性肾损伤更具敏感性和特异性的新型生物学标志物正越来越受到关注。     

早期尿生物标志物合并肾绞痛指数对重症监护病房儿童急性肾损伤的预测研究

目的:探讨早期尿生物标志物联合肾绞痛指数对重症监护病房儿童急性肾损伤(AKI)的预测能力。方法:选取肾绞痛并急性肾损伤患儿184例,确定患儿入院当天(Day0)的肾绞痛指数(RAI),由此判断是否对危重患儿第3天(Day3)持续、严重性急性肾损伤的发生具有预测作用(Day3-AKI:KDIGO Stage 2~3);同时研究RAI合并尿生物标志物是否对AKI 的预测具有优化作用。结果:184例患儿入院当天共有60例患儿(32.6%)出现肾绞痛(RAI≥8)。与Day0 非肾绞痛患儿相比,Day0 肾绞痛患儿的机械通气时间更长(P<0.01),器官衰竭时间更长(P<0.01),病死率更高(P<0.01)。Day3 共有15例患儿(9.6%)出现AKI,其中12例(80.0%)为Day0 肾绞痛患儿。尿中的生物标志物联合RAI可增加AKI预测的特异性和准确率,对Day3-AKI的预测具有明显的改善作用(P<0.01)。尿中性粒细胞明胶酶相关载脂蛋白的纳入可使Day3-AKI的RAI-ROC曲线下面积从0.80(95% CI:0.58,1.00)增加到0.97(95% CI:0.93,1.00)。结论:RAI可作为AKI危重患儿功能学危险分级的一种方法,为新型尿液标志物的测量和改善重度、持续性AKI的预测提供依据。     

FDA和EMEA批准7种肾损伤生物标志物正式使用

药物安全性预测联盟（PSTC）集合众多制药企业的力量,在美国食品药品监督局（FDA）和欧洲医药评价署（EMEA）的指导下致力于药物安全性监测的研究,并向FDA和EMEA呈交了7种肾损伤生物标志物的研究结果。目前,FDA和EMEA评估并接受了这7种新的生物标志物,使之用于检测尿中反映肾损伤的信号。这些新的检测指标现在可作为药物评价的一部分,用于实验室探索实验性药物的安全性,从而使药物在安全性上更加可靠。     

药物性肾损伤的早期发现及干预

药物性肾损伤起病隐匿,损伤早期为可逆反应,预后关键在于早期发现、早期干预。对就诊者建立完整的信息档案,临床医、护、药人员熟悉药物特性,因人而异个性化用药,用药前、中、后全过程定期检测早期肾损伤标志物,关注症状细节及时采取干预措施,以有效防止药物性肾损伤的发生发展。     

过敏性紫癜患儿早期肾损伤生物标志物特征分析

目的 分析过敏性紫癜患儿早期肾损伤相关生物标志物的临床特征,为临床诊治提供依据。方法 选择2015～2016年安徽省儿童医院收治的149例过敏性紫癜患儿,根据是否并发肾损害分为过敏性紫癜组（76例）和紫癜性肾炎组（73例）,分析两组患儿血清IgM、IgG、IgA和尿液β-N-乙酰氨基葡萄糖苷酶（NAG）、IgG、视黄醇结合蛋白（RBP）等指标的差异特征。结果 过敏性紫癜组患儿血清IgG阳性率（28.95%）、IgA阳性率（80.26%）高于紫癜性肾炎组,差异有统计学意义（P<0.05）;紫癜性肾炎组患儿尿液NAG、IgG、转铁蛋白（TRF）、微量白蛋白（MA）水平高于过敏性紫癜组,差异有统计学意义（P<0.05）。结论 血清IgA、IgG水平升高是过敏性紫癜发生风险的生物标志物,尿液NAG、IgG、TRF、MA水平差异对判断患儿早期肾损伤具有更明确的价值。     

尿液NGAL作为药物诱导急性肾毒性生物标志物的研究进展

寻找能早期、灵敏、特异地诊断肾毒性的新型生物标志物已成为目前药物临床前研究的热点。大量研究发现尿液中性粒细胞明胶酶相关脂质运载蛋白（NGAL）对肾损伤的诊断效能优于传统生物标志物血清肌酐和尿素氮。2018年尿液NGAL获得了美国食品药品监督管理局的批准,可用于I期临床试验中药物对肾小管损伤作用的监测,但其在临床前试验中的应用和推广仍有待进一步研究。就NGAL特性、作为药物诱导急性肾损伤生物标志物研究进展及其应用面临的挑战进行综述,以期为NGAL在我国新药研发中对急性肾毒性评价的应用奠定基础。     

Age-related differences in kidney injury biomarkers induced by cisplatin

Acute kidney injury (AKI) occurs in a half of cisplatin (CDDP)-treated patients. Traditional biomarkers including blood urea nitrogen (BUN) and serum creatinine (SCr) are still used for detection of CDDP-induced AKI, but these biomarkers are not specific or sensitive. The aim of this study was to identify the specific and sensitive biomarkers against CDDP-induced renal injury between young (3-week-old) and old (20-week-old) rats. All animals were intraperitoneally injected once with CDDP (6 mg/kg). After 3 days, all animals were sacrificed and serum, urine, and kidney tissues were collected. Urinary and serum biomarkers as well as histological changes were measured. CDDP-induced proximal tubular damage was apparent from histopathological examination, being more severe in 3-week-old rats accompanied by increased number of TUNEL-positive apoptotic cells. This was associated with elevated urinary kidney injury molecule-1 (KIM-1), glutathione-S-transferase alpha (GST-α), vascular endothelial growth factor (VEGF), and tissue inhibitor of metalloproteinases-1 (TIMP-1). In contrast, the levels of neutrophil gelatinase-associated lipocalin (NGAL) and osteopontin were significantly increased in 20-week-old rats after CDDP treatment. These results indicate that the use of age-specific urinary biomarkers is necessary to diagnosis of CDDP-induced AKI. Especially, urinary KIM-1, GST-α, TIMP-1, and VEGF levels may help in the early diagnosis of young patients with CDDP-induced AKI.     

Urinary kidney biomarkers for early detection of nephrotoxicity in clinical drug development

Early detection of drug-induced kidney injury is vital in drug development. Generally accepted biomarkers such as creatinine and blood urea nitrogen (BUN) lack sensitivity and early injury responses are missed. Many new biomarkers to detect nephrotoxicity for pre-clinical utilization have been described and their use is adopted in regulatory guidelines. However, guidance on appropriate biomarkers for clinical trials is minimal. We provide an overview of potentially useful kidney biomarkers that can be used in clinical trials. This includes guidance to select biomarkers suitable to capture specific characteristics of the (expected) kidney injury. We conclude that measurement of urinary kidney injury marker-1 (KIM-1) serves many purposes and is often an appropriate choice. Cystatin C captures effects on glomerular filtration rate (GFR), but this marker should preferably be combined with more specific markers to localize the origin of the observed effect. Untoward effects on tubules can be captured relatively well with several markers. Direct detection of glomerular injury is currently impossible since specific biomarkers are lacking. Indirect assessment of toxic effects on glomeruli is possible by using carefully selected panels of other injury markers. We conclude that it is possible to obtain appropriate information on nephrotoxicity in clinical drug development by using carefully selected panels of injury markers and suggest that identification and validation of specific glomerular biomarkers could be of great value.     

Comparison of kidney injury molecule-1 and other nephrotoxicity biomarkers in urine and kidney following acute exposure to gentamicin, mercury, and chromium.

Sensitive biomarkers are needed to detect kidney injury at the earliest stages. The objective of this study was to determine whether the appearance of kidney injury molecule-1 (Kim-1) protein ectodomain in urine and kidney injury molecule-1/hepatitis A viral cellular receptor-1 (Kim-1/Havcr1) gene expression in kidney tissue may be more predictive of renal injury after exposure to nephrotoxicants when compared to traditionally used biomarkers. Male Sprague-Dawley rats were injected with a range of doses of gentamicin, mercury (Hg; HgCl2), or chromium (Cr; K2Cr2O7). The results showed that increases in urinary Kim-1 and kidney Kim-1/Havcr1 gene expression paralleled the degree of severity of renal histopathology and were detected at lower doses of nephrotoxicants when compared to blood urea nitrogen (BUN), serum creatinine, and urinary N-acetyl-beta-D-glucosaminidase (NAG). In a time course study, urinary Kim-1 was elevated within 24 h after exposure to gentamicin (100 mg/kg), Hg (0.25 mg/kg), or Cr (5 mg/kg) and remained elevated through 72 h. NAG responses were nephrotoxicant dependent with elevations occurring early (gentamicin), late (Cr), or no change (Hg). At 72 h, after treatment with any of the three nephrotoxicants, there was increased Kim-1 immunoreactivity and necrosis involving approximately 50% of the proximal tubules; however, only urinary Kim-1 was significantly increased, while BUN, serum creatinine, and NAG were not different from controls. In rats treated with the hepatotoxicant galactosamine (1.1 mg/kg), serum alanine aminotransferase was increased, but no increase in urinary Kim-1 was observed. Urinary Kim-1 and kidney Kim-1/Havcr1 expression appear to be sensitive and tissue-specific biomarkers that will improve detection of early acute kidney injury following exposure to nephrotoxic chemicals and drugs.     

Time evolution of methotrexate‐induced kidney injury: A comparative study between different biomarkers of renal damage in rats

Methotrexate (MTX) is commonly used in the treatment of malignant diseases and autoimmune and chronic inflammatory disorders. Along with its effective therapeutic power, MTX has adverse effects on the kidneys. Discovery of new biomarkers is required to improve the early detection of renal damage and optimize the effectiveness of treatments. The aim of this study was to evaluate the time course of MTX‐induced nephrotoxicity and to compare the urinary excretion of the organic anion transporter 5 (uOat5) with alterations in other markers of renal function, and to elucidate the possible molecular mechanisms involved in uOat5. Animals were exposed to a unique dose of MTX (80 mg/kg body weight, intraperitoneal). Experiments were carried out at days 2, 4, 8 or 14 after MTX administration. Markers of renal damage, such as creatinine and urea plasma levels, urinary activity of alkaline phosphatase, microalbuminuria, urinary excretion of neutrophil gelatinase‐associated lipocalin (uNGAL) and histopathology, were evaluated. Renal organic anion transporter 5 (Oat5) expression and its presence in different urine fraction were assessed by western blotting. uOat5 was significantly increased 2 days after MTX treatment, before than any alteration in other parameters of kidney injury or renal morphology occurred. uNGAL showed an inverted pattern of urinary excretion compared to uOat5. Exosomal pathway is involved in the urinary excretion of Oat5 and depends on the degree of damage induced by MTX. These experimental data allow proposing uOat5 as a potential non‐invasive biomarker for early detection of MTX‐induced nephrotoxicity.     

KIM-1和 CysC在重症急性胰腺炎合并急性肾损伤中的早期诊断价值

目的：探讨尿液中肾损伤分子-1（KIM-1）和胱抑素C（CysC）在急性重症胰腺炎（SAP）合并急性肾损伤（AKI）患者中的早期诊断价值。方法收集15例健康体检者作为正常对照,将161例急性胰腺炎（AP）患者分为轻度（MAP）、中度（MSAP）、重度（SAP）3组,其中SAP组又分为AKI和非AKI亚组。采用酶联免疫分析法（ELISA法）测定正常对照组及AP患者发病后12～24 h的尿液KIM-1、CysC水平。结果 SAP组中AKI 亚组的KIM-1和CysC水平明显高于非AKI 亚组（P＜0．01）。结论尿KIM-1和CysC可以作为检测SAP合并AKI的早期生物标记物。     

肾损伤分子1对顺铂诱导的大鼠急性肾损伤的预测研究

目的 对肾损伤分子1 (KIM-1)预测顺铂诱导的大鼠急性肾损伤进行研究.方法 以顺铂为工具药,诱导大鼠急性肾损伤模型.采集尿样、肾组织样本,对肾组织样本进行病理切片检查,以确定造模是否成功.用ELISA法测定尿样中KIM-1蛋白含量;RT-PCR法检测大鼠肾脏KIM-1基因表达水平;Western blotting法检测大鼠肾组织中KIM-1的蛋白含量,并通过免疫组化法定性定位分析大鼠肾组织中KIM-1蛋白表达情况,以确定急性肾损伤发生时KIM-1是否可以作为敏感的检测指标.结果 当模型组大鼠肾脏皮髓交界处出现程度不等的肾小管扩张,肾小管上皮细胞变性、坏死脱落,基底膜裸露等病理改变时,ELISA法检测尿KIM-1,RT-PCR法和Western blotting法检测肾组织中的KIM-1表达水平均明显升高,免疫组化显示模型组所有大鼠在肾皮髓交界部位可见大量的染色阳性的肾小管,该染色阳性区域与组织病理学检查中的异常肾小管分布区域基本一致.结论 KIM-1可作为一种敏感的生物标志物对顺铂诱导的大鼠急性肾损伤进行预测.     

Use of a glyphosate-based herbicide-induced nephrotoxicity model to investigate a panel of kidney injury biomarkers

Accidental or intentional ingestion of glyphosate surfactant-based herbicides, like Roundup^®, leads to nephrotoxicity as well as death. In this study, a panel of kidney injury biomarkers was evaluated in terms of suitability to detect acute kidney injury and dysfunction. The Roundup^® intoxication model involved oral administration of glyphosate to rats at dose levels of 250, 500, 1200 and 2500 mg/kg. Urinary and plasma biomarker patterns were investigated at 8, 24 and 48 h after dosing. Biomarkers were quantified by absolute concentration; by normalising to urine creatinine; and by calculating the excretion rate. The diagnostic performances of each method in predicting of acute kidney injury were compared. By Receiver Operating Characteristic (ROC) analysis of the selected biomarkers, only urinary kidney injury molecule-1 (KIM-1) best predicted histological changes at 8 h (best cut-off point > 0.00029 μg/ml). Plasma creatinine performed better than other biomarkers at 24 h (best cut-off point > 0.21 mg/dl). Urinary KIM-1 was the best early biomarker of kidney injury in this glyphosate-induced nephrotoxicity model.     

Usefulness of urinary kidney injury molecule‐1 (Kim‐1) as a biomarker for cisplatin‐induced sub‐chronic kidney injury

We explored biomarkers suitable for monitoring sub‐chronic kidney injury using the three rat models of cisplatin (CDDP)‐induced kidney injury, which were designed to extend the current knowledge beyond the sub‐acute exposure period. In the pilot study, a single intravenous administration of 1.5 mg kg^–1 CDDP to rats was confirmed to result in no histopathological changes. Subsequently, CDDP was intravenously administered to rats at a dose of 1.5 mg kg^–1 for 4 days at 24‐h intervals (Experimental model 1) and for up to 10 weeks at weekly intervals (Experimental models 2 and 3), and the changes in blood and urine components, such as recently recommended urinary biomarkers (Kim‐1, clusterin and so on) and traditional blood biomarkers (blood urea nitrogen and serum creatinine), were examined together with the histopathological changes in renal tissues during the development of the kidney injury in each model. In these experimental models, a significant increase in urinary Kim‐1 was observed prior to the histopathological changes in renal tissues, and these changes were retained after the adverse histopathological changes. Significant changes in all of the other urinary biomarkers examined occurred along with the histopathological changes. In addition, the increase in urinary Kim‐1 after weekly treatment with CDDP for 4 weeks was reduced in a time‐dependent manner after cessation of the drug. The present findings indicate that urinary Kim‐1 is the most useful biomarker for CDDP‐induced rat sub‐chronic kidney injury among the biomarkers examined. Copyright © 2014 John Wiley & Sons, Ltd.