Effect of prednisolone on glyoxalase 1 in an inbred mouse model of aristolochic acid nephropathy using a proteomics method with fluorogenic derivatization-liquid chromatography-tandem mass spectrometry

RESEARCH ARTICLE Effect of prednisolone on glyoxalase 1 in an inbred mouse model of aristolochic acid nephropathy using a proteomics method with fluorogenic derivatization-liquid chromatography-tandem mass spectrometry Shih-Ming Chen ID1¤*, Chia-En Lin1¤, Hung-Hsiang Chen1¤, Yu-Fan Cheng1¤, HuiWen Cheng1¤, Kazuhiro Imai2 a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 1 School of Pharmacy, Taipei Medical University, Taipei, Taiwan, 2 Research Institute of Pharmaceutical Sciences, Musashino University, Tokyo, Japan ¤ Current address: Taipei Medical University, Taipei, Taiwan * Abstract OPEN ACCESS Citation: Chen S, Lin C, Chen H-H, Cheng Y-F, Cheng H-W, Imai K (2020) Effect of prednisolone on glyoxalase 1 in an inbred mouse model of aristolochic acid nephropathy using a proteomics method with fluorogenic derivatization-liquid chromatography-tandem mass spectrometry. PLoS ONE 15(1): e0227838. https://doi.org/ 10.1371/journal.pone.0227838 Editor: Fabio Sallustio, University of Bari Aldo Moro, ITALY Received: October 4, 2019 Accepted: December 30, 2019 Published: January 22, 2020 Peer Review History: PLOS recognizes the benefits of transparency in the peer review process; therefore, we enable the publication of all of the content of peer review and author responses alongside final, published articles. The editorial history of this article is available here: https://doi.org/10.1371/journal.pone.0227838 Copyright: © 2020 Chen et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Prednisolone is involved in glucose homeostasis and has been used for treatment for aristolochic acid (AA) nephropathy (AAN), but its effect on glycolysis in kidney has not yet been clarified. This study aims to investigate the effect in terms of altered proteins after prednisolone treatment in a mice model of AAN using a proteomics technique. The six-week C3H/He female mice were administrated AA (0.5 mg/kg/day) for 56 days. AA+P group mice were then given prednisolone (2 mg/kg/day) via oral gavage for the next 14 days, and AA group mice were fed water instead. The tubulointerstitial damage was improved after prednisolone treatment comparing to that of AA group. Kidney homogenates were harvested to perform the proteomics analysis with fluorogenic derivatization-liquid chromatography-tandem mass spectrometry method (FD-LC-MS/MS). On the other hand, urinary methylglyoxal and D-lactate levels were determined by high performance liquid chromatography with fluorescence detection. There were 47 altered peaks and 39 corresponding proteins on day 14 among the groups, and the glycolysis-related proteins, especially glyoxalase 1 (GLO1), fructosebisphosphate aldolase B (aldolase B), and triosephosphate isomerase (TPI), decreased in the AA+P group. Meanwhile, prednisolone decreased the urinary amount of methylglyoxal (AA+P: 2.004 ± 0.301 μg vs. AA: 2.741 ± 0.630 μg, p < 0.05), which was accompanied with decrease in urinary amount of D-lactate (AA+P: 54.07 ± 5.45 μmol vs. AA: 86.09 ± 8.44 μmol, p < 0.05). Prednisolone thus alleviated inflammation and interstitial renal fibrosis. The renal protective mechanism might be associated with down-regulation of GLO1 via reducing the contents of methylglyoxal derived from glycolysis. With the aid of proteomics analysis and the determination of methylglyoxal and its metabolite-D-lactate, we have demonstrated for the first time the biochemical efficacy of prednisolone, and urinary methylglyoxal and its metabolite-D-lactate might be potential biomarkers for AAN. PLOS ONE | https://doi.org/10.1371/journal.pone.0227838 January 22, 2020 1 / 19 Proteomics analysis: Effect of prednisolone on glyoxalase 1 in aristolochic acid nephropathy using FD-LC-MS/MS Data Availability Statement: All relevant data are within the manuscript and its Supporting Information files. Funding: We are grateful to the financial support from the Cathay General Hospital (108CGH-TMU06). Competing interests: The authors have declared that no competing interests exist. Introduction Aristolochic acid nephropathy (AAN) was first introduced in 1993 [1]. After the Belgian women ingested slimming pills containing aristolochic acid (AA), their renal function dramatically decreased and managed by dialysis. The feature of AAN is rapid progression into interstitial renal fibrosis and end-stage renal disease [1, 2]. AA is extracted from the Aristolochiaceae species and was used for anti-inflammatory activities in traditional medicine. AA is a mixture of 8-methoxy-6-nitrophenanthro-(3,4-D)-1,3-dioxolo-5-carboxylic acid (aristolochic acid I [AAI]) and its 8-demethoxylated form (aristolochic acid II [AAII]) [3]. AAI shows stronger nephrotoxicity than AAII in AAN because of the O-methoxy group at position C-8 of the nitrophenanthrene ring [4]. This structure of AAI facilitates AA interactions with DNA, and this AA-DNA adduct leads to cytotoxicity and carcinogenicity. A recent study indicates that AA-induced upper tract urothelial cancer is related to p38 and extracellular signal regulated kinases (ERK) sub-pathways [5]. Despite prohibition of AAcontaining herbs, patients are still suffering from AAN [6]. Vanherweghem et al. successfully treated AAN with prednisolone, because AAN is thought to be related to the immune response, such as infiltration of immune cells into the renal cortex [7]. Recently, Ma et al. showed that low-dose prednisone (0.5 mg/kg) is effective at slowing the progression of AAN via the suppression of monocyte chemoattractant protein-1 (MCP-1) and transforming growth factor-β (TGF-β) activities [8]. Prednisolone, one of the glucocorticoids, acts as endogenous cortisol that is released from the hypothalamic-pituitary-adrenal (HPA) axis, particularly in the case of stress or injury [9]. Prednisolone fights inflammation via the regulation of tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1), and interleukin-6 (IL-6) etc, thus, widely applied to treating various inflammation diseases or states, such as allergies, asthma, dermatitis, rheumatic disorders, systemic lupus erythematosus, and autoimmune disorders[10–12]. Recently, Baudoux et al. demonstrated that cluster of differentiation CD4+ and CD8+ T-cells regulate immune responses in AA-induced acute tubular necrosis [13]. However, the mechanism of prednisolone treatment for AAN is still unclear. Moreover, prednisolone plays an important role in glucose homeostasis, but the relationship between prednisolone and glycolysis, including the impact of methylglyoxal, a by-product of glycolysis, has never been studied. The proteomics study with fluorogenic derivatization-liquid chromatography/tandem mass spectrometry (FD-LC-MS/ MS) was introduced in 2004 [14, 15]. This FD-LC-MS/MS method was widely applied to screening proteins in cell lines [16], ra (...truncated)