NGAL蛋白

ＮＧＡＬ(ｎｅｕｔｒｏｐｈｉｌｇｅｌａｔｉｎａｓｅ ａｓｓｏｃｉａｔｅｄｌｉｐｏｃａｌｉｎ)是ｌｉｐｏｃａｌｉｎ家族的一个新成员 ,是Ｋｊｅｌｄｓｅｎ等人[1] 在1993年研究中性粒细胞 92ｋＤ的明胶酶 ,即ＭＭＰ 9(基质金属蛋白酶 9)时发现的。随后 ,ＮＧＡＬｃＤＮＡ及其全基因序列分别于 1994和 1997年被克隆鉴定[2 ,3] 。已知ＮＧＡＬ蛋白具有运输疏水性小分子 ,调节ＭＭＰ 9活性等功能。近年来 ,随着ＮＧＡＬ蛋白在人体组织细胞中广泛分布情况的认证[4 ] ,特别是在一些肿瘤组织细胞中出现该基因异常过表达等事实的认识[5] ,有关Ｎ…     

SHEEC食管癌细胞中NGAL基因的功能

中性粒细胞明胶酶相关脂质运载蛋白 (neutrophilgelatinase associatedlipocalin ,NGAL)是脂质运载蛋白(lipocalin)家族的一个新成员 ,可能是人类的一种新的癌基因 ,但是在肿瘤中的功能不清楚。以往研究发现NGAL基因在SHEEC食管癌细胞中显著过表达 ,表明该细胞是一种用来揭示NGAL基因在肿瘤中功能的良好模型。采用反义封闭技术 ,同时结合裸鼠成瘤实验等研究了反义封闭NGAL基因转录对SHEEC食管癌细胞的浸润和分裂增殖等行为的影响。结果发现 ,反义封闭NGAL基因转录不但可以有效地降低SHEEC细胞分泌的基质金属蛋白酶 9和基质金属蛋白酶 2的活性 ,而与此同时裸鼠成瘤细胞的浸润行为也相应地受到了明显抑制 ,然而SHEEC细胞端粒的长度、拓扑异构酶II的含量以及细胞增殖指数等未发生明显变化。表明NGAL基因在食管癌细胞SHEEC中的功能可能主要是通过明胶酶在促进肿瘤细胞的浸润中发挥作用 ,而可能与肿瘤细胞分裂增殖的相关性不明显     

Neutrophil Gelatinase-associated Lipocalin in Normal and Neoplastic Human Tissues. Cell Type-specific Pattern of Expression

Neutrophil gelatinase-associated lipocalin (NGAL) has recently been identified in myeloperoxidase-negative neutrophil granules. Members of the lipocalin family are thought to bind and transport small lipophilic molecules such as retinoids and roles in cell regulation have been proposed. Recently, NGAL has also been demonstrated in the colonic mucosa in certain pathologic conditions.The aim of this study was to examine the distribution of NGAL in normal and neoplastic tissues by immunohistochemistry. Interestingly, NGAL was found in a variety of normal and pathological human tissues. A cell type-specific pattern of expression was seen in bronchus, stomach, small intestine, pancreas, kidney, prostate gland, and thymus. The comparative analysis of the putative rat homologue neu-related lipocalin showed a very similar pattern of expression with the exception of pancreas and kidney. Neoplastic human tissues showed a very heterogeneous expression of NGAL protein. High NGAL levels were found in adenocarcinomas of lung, colon and pancreas. In contrast, renal cell carcinomas of various subtypes and prostate cancers contained low NGAL levels. Lymphomas and thymic tumours were negative for NGAL immuno-labeling. Knowledge about the location of NGAL in normal cells and in disease states provides the first clues towards understanding its biological function.     

建立NGAL诊断阈值有利于临床判断和发现急性肾功能损伤

为探讨中性粒细胞相关载脂蛋白(neutrophil gelatinase-associated lipocalin, NGAL)检测在肝胆疾病患者继发急性肾功能损伤(acute kidney injury, AKI)中的诊断性能,本研究回顾性收集了476例肝胆疾病患者与225例健康受试者作为实验组,根据血清中性粒细胞相关载脂蛋白(NGAL)、血清肌酐(serum creatinine,s Cr)、尿素(Urea)、半胱氨酸蛋白酶抑制剂C (cystatin C, CysC)、估算肾小球滤过率(estimating glomerular filtration rate, eGFR)水平和尿量,分为急性肾功能损伤(AKI)组和高风险(high-risk, HR)组、低风险(low-risk,LR)组和对照(health control, HC)组,进而建立上述血清指标在急性肾功能损伤(AKI)诊断性能最大时的判断界值;随后,选取145例肝胆疾病患者作为验证组,以评估各观察指标对肝胆疾病继发急性肾功能损伤(AKI)的诊断性能。结果表明:本实验组中各亚组间所观察指标之血清水平均有统计学差异(p<0.05)。中性粒细胞相关载脂蛋白(NGAL)诊断性能最大时的判断界值为205.2μg/L。验证组患者各指标阳性检出率,在低风险(LR)亚组中中性粒细胞相关载脂蛋白(NGAL)均高于其它指标(p<0.05),但在高风险(HR)亚组中中性粒细胞相关载脂蛋白(NGAL)仅高于血清肌酐(sCr)(p<0.05),而与半胱氨酸蛋白酶抑制剂C (CysC)和估算肾小球滤过率(eGFR)无统计学差异(p>0.05);再经分层风险分析各指标预测高风险(HR)亚组患者急性肾功能损伤(AKI)发生的能力,中性粒细胞相关载脂蛋白(NGAL)的优势比(OR=21.0 (2.3, 192.8)),是半胱氨酸蛋白酶抑制剂C(CysC)或估算肾小球滤过率(eGFR)(OR (95%CI)=3.3 (0.7, 15.3))的6.4倍。本研究初步结论表明,肝胆疾病患者诊断急性肾功能损伤(AKI)时应考虑中性粒细胞相关载脂蛋白(NGAL)肝脏合成代谢的作用。因此实验室应建立适宜的中性粒细胞相关载脂蛋白(NGAL)诊断阈值以有利于临床准确判断及早期发现急性肾功能损伤(AKI)。     

血浆中性粒细胞明胶酶相关脂质运载蛋白(NGAL)的变化与婴幼儿先心病术后急性肺损伤的关系

目的:明确血浆中性粒细胞明胶酶相关脂质运载蛋白(NGAL)的变化能否预测婴幼儿先心病术后急性肺损伤(ALI)的发生。方法:纳入2013年10月至2014年5月年龄≤36月的先心病合并肺动脉高压的婴幼儿患者的相关资料,检测体外循环(CPB)术后2 h、12 h、24 h、48 h和72 h的血浆NGAL水平,分析NGAL与术后发生ALI之间的关系。结果:本研究共纳入患者61例,术后发生ALI患者9例(14.8%)。所有患者CPB术后5个时间点的变化趋势为:CPB术后2 h NGAL浓度开始升高,均值为27.28μg/L,至12 h达到最高峰,均值为30.61μg/L,24 h开始下降;其中12 h、24 h和72 h与前一时间点比较均具有显著性差异(P0.05)。进一步分组显示,NGAL浓度变化趋势主要在ALI组:ALI组患者12 h、24 h、48 h和72 h的NGAL浓度与前一个时间点比较,变化均具有显著差异(P0.05),NGAL峰值浓度出现在12 h,其均值为40.82μg/L;而无ALI组患者NGAL浓度只在24 h和12 h之间比较有显著差异(P0.05);ALI组患者5个时间点的NGAL浓度均比无ALI组患者高,具有显著性差异(P0.05)。结论:血浆NGAL在早期预测婴幼儿体外循环术后发生ALI可能具有重要作用。     

Biochemical and Structural Characterization of the Interaction between the Siderocalin NGAL/LCN2 (Neutrophil Gelatinase-associated Lipocalin/Lipocalin 2) and the N-terminal Domain of Its Endocytic Receptor SLC22A17

The neutrophil gelatinase-associated lipocalin (NGAL, also known as LCN2) and its cellular receptor (LCN2-R, SLC22A17) are involved in many physiological and pathological processes such as cell differentiation, apoptosis, and inflammation. These pleiotropic functions mainly rely on NGAL''s siderophore-mediated iron transport properties. However, the molecular determinants underlying the interaction between NGAL and its cellular receptor remain largely unknown. Here, using solution-state biomolecular NMR in conjunction with other biophysical methods, we show that the N-terminal domain of LCN2-R is a soluble extracellular domain that is intrinsically disordered and interacts with NGAL preferentially in its apo state to form a fuzzy complex. The relatively weak affinity (≈10 μm) between human LCN2-R-NTD and apoNGAL suggests that the N terminus on its own cannot account for the internalization of NGAL by LCN2-R. However, human LCN2-R-NTD could be involved in the fine-tuning of the interaction between NGAL and its cellular receptor or in a biochemical mechanism allowing the receptor to discriminate between apo- and holo-NGAL.     

MicroRNA-138 Suppresses Neutrophil Gelatinase-Associated Lipocalin Expression and Inhibits Tumorigenicity

The expression of neutrophil gelatinase-associated lipocalin (NGAL) is up-regulated in some cancers; therefore NGAL has potential as a tumor biomarker. Although the regulation mechanism for this is unknown, one study has shown that it is likely to involve a microRNA (miRNA). Here, we investigate the relation between miRNA expression and NGAL expression, and the role of NGAL in tumorigenesis. Using miRNA target–detecting software, we analyze the mRNA sequence of NGAL and identify a target site for microRNA-138 (miR-138) in nucleotides 25–53 of the 3′ UTR. We then analyze NGAL and miR-138 expression in three cancer cell lines originating from breast, endometrial and pancreatic carcinomas (the MCF-7, RL95-2 and AsPC-1 cell lines), respectively, using quantitative (real-time) PCR and western blot analysis. Metastasis is a critical event in cancer progression, in which malignant cell proliferation, migration and invasion increase. To determine whether miR-138-regulated NGAL expression is associated with metastasis, the proliferation and migration of the cell line are examined after miR-138 transfection. Using nude mice, we examine both the tumorigenicity of these cell lines and of miR-138-transfected cancer cells in vivo, as well as the effect of treating tumors with an antibody against NGAL. Our results show that these cancer cell lines down-regulate NGAL when miR-138 is highly expressed. Ectopic transfection of miR-138 suppresses NGAL expression and cell migration in RL95-2 and AsPC-1 cells, demonstrating that miR-138-regulated NGAL expression is associated with cell migration. Additionally, injection of the NGAL antibody diminishes NGAL-mediated tumorigenesis in nude mice, and miR-138 transfection of cancer cells reduces tumor formation. As the cell proliferation data showed that the tumor size should be regulated by NGAL-related cell growth. Taken together, our results indicate that NGAL may be a good target for cancer therapy and suggest that miR-138 acts as a tumor suppressor and may prevent metastasis.     

The Solution Structure,Binding Properties,and Dynamics of the Bacterial Siderophore-binding Protein FepB

The periplasmic binding protein (PBP) FepB plays a key role in transporting the catecholate siderophore ferric enterobactin from the outer to the inner membrane in Gram-negative bacteria. The solution structures of the 34-kDa apo- and holo-FepB from Escherichia coli, solved by NMR, represent the first solution structures determined for the type III class of PBPs. Unlike type I and II PBPs, which undergo large “Venus flytrap” conformational changes upon ligand binding, both forms of FepB maintain similar overall folds; however, binding of the ligand is accompanied by significant loop movements. Reverse methyl cross-saturation experiments corroborated chemical shift perturbation results and uniquely defined the binding pocket for gallium enterobactin (GaEnt). NMR relaxation experiments indicated that a flexible loop (residues 225–250) adopted a more rigid and extended conformation upon ligand binding, which positioned residues for optimal interactions with the ligand and the cytoplasmic membrane ABC transporter (FepCD), respectively. In conclusion, this work highlights the pivotal role that structural dynamics plays in ligand binding and transporter interactions in type III PBPs.     

Ir-LBP,an Ixodes ricinus Tick Salivary LTB4-Binding Lipocalin,Interferes with Host Neutrophil Function

Background

During their blood meal, ticks secrete a wide variety of proteins that can interfere with their host''s defense mechanisms. Among these proteins, lipocalins play a major role in the modulation of the inflammatory response.

Methodology/Principal Findings

We previously identified 14 new lipocalin genes in the tick Ixodes ricinus. One of them codes for a protein that specifically binds leukotriene B4 with a very high affinity (Kd: ±1 nM), similar to that of the neutrophil transmembrane receptor BLT1. By in silico approaches, we modeled the 3D structure of the protein and the binding of LTB4 into the ligand pocket. This protein, called Ir-LBP, inhibits neutrophil chemotaxis in vitro and delays LTB4-induced apoptosis. Ir-LBP also inhibits the host inflammatory response in vivo by decreasing the number and activation of neutrophils located at the tick bite site. Thus, Ir-LBP participates in the tick''s ability to interfere with proper neutrophil function in inflammation.

Conclusions/Significance

These elements suggest that Ir-LBP is a “scavenger” of LTB4, which, in combination with other factors, such as histamine-binding proteins or proteins inhibiting the classical or alternative complement pathways, permits the tick to properly manage its blood meal. Moreover, with regard to its properties, Ir-LBP could possibly be used as a therapeutic tool for illnesses associated with an increased LTB4 production.     

Complementing the Eukaryotic Protein Interactome

Protein interaction networks are important for the understanding of regulatory mechanisms, for the explanation of experimental data and for the prediction of protein functions. Unfortunately, most interaction data is available only for model organisms. As a possible remedy, the transfer of interactions to organisms of interest is common practice, but it is not clear when interactions can be transferred from one organism to another and, thus, the confidence in the derived interactions is low. Here, we propose to use a rich set of features to train Random Forests in order to score transferred interactions. We evaluated the transfer from a range of eukaryotic organisms to S. cerevisiae using orthologs. Directly transferred interactions to S. cerevisiae are on average only 24% consistent with the current S. cerevisiae interaction network. By using commonly applied filter approaches the transfer precision can be improved, but at the cost of a large decrease in the number of transferred interactions. Our Random Forest approach uses various features derived from both the target and the source network as well as the ortholog annotations to assign confidence values to transferred interactions. Thereby, we could increase the average transfer consistency to 85%, while still transferring almost 70% of all correctly transferable interactions. We tested our approach for the transfer of interactions to other species and showed that our approach outperforms competing methods for the transfer of interactions to species where no experimental knowledge is available. Finally, we applied our predictor to score transferred interactions to 83 targets species and we were able to extend the available interactome of B. taurus, M. musculus and G. gallus with over 40,000 interactions each. Our transferred interaction networks are publicly available via our web interface, which allows to inspect and download transferred interaction sets of different sizes, for various species, and at specified expected precision levels. Availability: http://services.bio.ifi.lmu.de/coin-db/.     

脂质运载蛋白突变文库的应用及研究现状

脂质运载蛋白（1ipocaan）蛋白家族是一类广泛存在于生物界、与体内小分子物质运输和贮存相关的蛋白质家族。由于其高级结构具有与抗体高级结构类似的特点：由保守区和超变区组成,因此,以脂质运载蛋白为骨架构建的突变文库也成为一个新的研究领域,该文介绍脂质运载蛋白家族的结构特点、脂质运载蛋白突变文库的应用及研究现状。     

Plasma Neutrophil Gelatinase-Associated Lipocalin Is Primarily Related to Inflammation during Sepsis: A Translational Approach

Acute kidney injury (AKI) during sepsis is common and underestimated. Plasma neutrophil gelatinase-associated lipocalin (plasma-NGAL) is discussed as new biomarker for AKI diagnosis, but during inflammation its function and diagnostic impact remain unclear. The association between plasma-NGAL and inflammatory markers in septic patients, but also in healthy controls and patients with chronic inflammation before and after either maximum exercise test or treatment with an anti-TNF therapy were investigated. In-vitro blood stimulations with IL-6, lipopolysaccharide, NGAL or its combinations were performed to investigate cause-effect-relationship. Plasma-NGAL levels were stronger associated with inflammation markers including IL-6 (Sepsis: r=0.785 P<0.001; chronic inflammation after anti-TNF: r=0.558 P<0.001), IL-8 (Sepsis: r=0.714 P<0.004; healthy controls after exercise r=0.786 P<0.028; chronic inflammation before anti-TNF: r=0.429 P<0.041) and IL-10 (healthy controls before exercise: r=0.791 P<0.028) than with kidney injury or function. Correlation to kidney injury or function was found only in septic patients (for creatinine: r= 0.906 P<0.001; for eGFR: r= -0.686 P=0.005) and in patients with rheumatic disease after anti-TNF therapy (for creatinine: r= 0.466 P<0.025). In stimulation assays with IL-6 and lipopolysaccharide plasma-NGAL was increased. Co-stimulation of lipopolysaccharide with plasma-NGAL decreased cellular injury (P<0.05) and in trend IL-10 levels (P=0.057). Septic mice demonstrated a significantly improved survival rate after NGAL treatment (P<0.01). Plasma-NGAL seams to be strongly involved in inflammation. For clinical relevance, it might not only be useful for AKI detection during severe inflammation - indeed it has to be interpreted carefully within this setting - but additionally might offer therapeutic potential.     

Association between Plasma Neutrophil Gelatinase Associated Lipocalin Level and Obstructive Sleep Apnea or Nocturnal Intermittent Hypoxia

Background

Both obstructive sleep apnea (OSA) and a novel lipocalin, neutrophil gelatinase associated lipocalin (Ngal), have been reported to be closely linked with cardiovascular disease and loss of kidney function through chronic inflammation. However, the relationship between OSA and Ngal has never been investigated.

Objectives

To evaluate the relationship between Ngal and OSA in clinical practice.

Methods

In 102 patients, polysomnography was performed to diagnose OSA and plasma Ngal levels were measured. The correlations between Ngal levels and OSA severity and other clinical variables were evaluated. Of the 46 patients who began treatment with continuous positive airway pressure (CPAP), Ngal levels were reevaluated after three months of treatment in 25 patients.

Results

The Ngal level correlated significantly with OSA severity as determined by the apnea hypopnea index (r = 0.24, p = 0.01) and 4% oxygen desaturation index (ODI) (r = 0.26, p = 0.01). Multiple regression analysis showed that the Ngal level was associated with 4%ODI independently of other clinical variables. Compliance was good in 13 of the 25 patients who used CPAP. Although the OSA (4%ODI: 33.1±16.7 to 1.1±1.9/h, p<0.01) had significantly improved in those with good compliance, the Ngal levels were not significantly changed (60.5±18.1 before CPAP vs 64.2±13.9 ng/ml after CPAP, p = 0.27).

Conclusions

Plasma Ngal levels were positively associated with the severity of OSA. However, the contribution rate of OSA to systemic Ngal secretion was small and changes in Ngal levels appeared to be influenced largely by other confounding factors. Therefore, it does not seem reasonable to use the Ngal level as a specific biomarker of OSA in clinical practice.     

Quality Control in Eukaryotic Membrane Protein Overproduction

The overexpression of authentically folded eukaryotic membrane proteins in milligramme quantities is a fundamental prerequisite for structural studies. One of the most commonly used expression systems for the production of mammalian membrane proteins is the baculovirus expression system in insect cells. However, a detailed analysis by radioligand binding and comparative Western blotting of G protein-coupled receptors and a transporter produced in insect cells showed that a considerable proportion of the expressed protein was misfolded and incapable of ligand binding. In contrast, production of the same membrane proteins in stable inducible mammalian cell lines suggested that the majority was folded correctly. It was noted that detergent solubilisation of the misfolded membrane proteins using either digitonin or dodecylmaltoside was considerably less efficient than using sodium dodecyl sulfate or foscholine-12, whilst these detergents were equally efficient at solubilising correctly folded membrane proteins. This provides a simple and rapid test to suggest whether heterologously expressed mammalian membrane proteins are indeed correctly folded, without requiring radioligand binding assays. This will greatly facilitate the high-throughput production of fully functional membrane proteins for structural studies.     

Urine/Plasma Neutrophil Gelatinase Associated Lipocalin Ratio Is a Sensitive and Specific Marker of Subclinical Acute Kidney Injury in Mice

Background

Detection of acute kidney injury (AKI) is still a challenge if conventional markers of kidney function are within reference range. We studied the sensitivity and specificity of NGAL as an AKI marker at different degrees of renal ischemia.

Methods

Male C57BL/6J mice were subjected to 10-, 20- or 30-min unilateral renal ischemia, to control operation or no operation, and AKI was evaluated 1 day later by histology, immunohistochemistry, BUN, creatinine, NGAL (plasma and urine) and renal NGAL mRNA expression.

Results

A short (10-min) ischemia did not alter BUN or kidney histology, but elevated plasma and urinary NGAL level and renal NGAL mRNA expression although to a much smaller extent than longer ischemia. Surprisingly, control operation elevated plasma NGAL and renal NGAL mRNA expression to a similar extent as 10-min ischemia. Further, the ratio of urine to plasma NGAL was the best parameter to differentiate a 10-min ischemic injury from control operation, while it was similar in the non and control-operated groups.

Conclusions

These results suggest that urinary NGAL excretion and especially ratio of urine to plasma NGAL are sensitive and specific markers of subclinical acute kidney injury in mice.     

Urinary Neutrophil Gelatinase-Associated Lipocalin: A Useful Biomarker for Tacrolimus-Induced Acute Kidney Injury in Liver Transplant Patients

Tacrolimus is widely used as an immunosuppressant in liver transplantation, and tacrolimus-induced acute kidney injury (AKI) is a serious complication of liver transplantation. For early detection of AKI, various urinary biomarkers such as monocyte chemotactic protein-1, liver-type fatty acid-binding protein, interleukin-18, osteopontin, cystatin C, clusterin and neutrophil gelatinase-associated lipocalin (NGAL) have been identified. Here, we attempt to identify urinary biomarkers for the early detection of tacrolimus-induced AKI in liver transplant patients. Urine samples were collected from 31 patients after living-donor liver transplantation (LDLT). Twenty recipients developed tacrolimus-induced AKI. After the initiation of tacrolimus therapy, urine samples were collected on postoperative days 7, 14, and 21. In patients who experienced AKI during postoperative day 21, additional spot urine samples were collected on postoperative days 28, 35, 42, 49, and 58. The 8 healthy volunteers, whose renal and liver functions were normal, were asked to collect their blood and spot urine samples. The urinary levels of NGAL, monocyte chemotactic protein-1 and liver-type fatty acid-binding protein were significantly higher in patients with AKI than in those without, while those of interleukin-18, osteopontin, cystatin C and clusterin did not differ between the 2 groups. The area under the receiver operating characteristics curve of urinary NGAL was 0.876 (95% confidence interval, 0.800–0.951; P<0.0001), which was better than those of the other six urinary biomarkers. In addition, the urinary levels of NGAL at postoperative day 1 (p = 0.0446) and day 7 (p = 0.0006) can be a good predictive marker for tacrolimus-induced AKI within next 6 days, respectively. In conclusion, urinary NGAL is a sensitive biomarker for tacrolimus-induced AKI, and may help predict renal event caused by tacrolimus therapy in liver transplant patients.     

骨形成蛋白诱导型真核载体的构建及其表达

成骨分化相关基因骨钙素 (OC)等的启动子内均含有成骨特异性转录因子Cbfa1特异性作用元件 ,而骨形成蛋白 (bonemorphogeneticprotein ,BMP)的促成骨分化作用正是通过其首先引起Cbfa1的升高 ,而后Cbfa1激活这些基因的表达 ,最终出现成骨分化表型 .为解决BMP没有理想的活性测定方法的问题 ,在RT PCR结果证实BMP 2可促进NIH3T3和C2C12细胞Cbfa1表达后 ,构建了串联6个Cbfa1作用元件的小鼠OC部分启动子 (6OCP)控制萤光素酶 (luciferase)报告基因的真核表达质粒 ,以期来放大BMP诱导报告基因表达的作用效果 .即通过细胞转染、rhBMP 2刺激后检测萤光素酶活性变化 ,从而间接定量测定rhBMP 2的生物学活性 .结果表明 ,pcDNA3 6OCP Luc转染细胞后其报告基因的基础活性较pcDNA3 Luc大为降低 ;而且在一定剂量范围内 ,转染细胞的萤光素酶活性 (荧光值 )随rhBMP 2剂量增加而升高 ,并呈线性正相关 ,为建立BMP活性定量测定的方法打下基础     

Neutrophil Gelatinase Associated Lipocalin (NGAL) in Leptospirosis Acute Kidney Injury: A Multicenter Study in Thailand

AKI is one of the most serious complications of leptospirosis, an important zoonosis in the tropics. Recently, NGAL, one of the novel AKI biomarkers, is extensively studied in various specific settings such as sepsis, cardiac surgery, and radiocontrast nephropathy. In this multicenter study, we aimed to study the role of NGAL as an early marker and an outcome predictor of leptospirosis associated AKI. Patients who presented with clinical suspiciousness of leptospirosis were prospectively enrolled in 9 centers from August 2012 to November 2014. The first day of enrollment was the first day of clinical suspicious leptospirosis. Blood and urine samples were serially collected on the first three days and day 7 after enrollment. We used three standard techniques (microscopic agglutination test, direct culture, and PCR technique) to confirm the diagnosis of leptospirosis. KDIGO criteria were used for AKI diagnosis. Recovery was defined as alive and not requiring dialysis during hospitalization or maintaining maximum KDIGO stage at hospital discharge. Of the 221 recruited cases, 113 cases were leptospirosis confirmed cases. Thirty seven percent developed AKI. Median uNGAL and pNGAL levels in those developing AKI were significantly higher than in patients not developing AKI [253.8 (631.4) vs 24.1 (49.6) ng/ml, p < 0.001] and [1,030 (802.5) vs 192.0 (209.0) ng/ml, p < 0.001], respectively. uNGAL and pNGAL levels associated with AKI had AUC-ROC of 0.91, and 0.92, respectively. Both of urine NGAL and pNGAL level between AKI-recovery group and AKI-non recovery were comparable. From this multicenter study, uNGAL and pNGAL provided the promising result to be a marker for leptospirosis associated AKI. However, both of them did not show the potential role to be the predictor of renal recovery in this specific setting.