Summary of Study ST002532
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001628. The data can be accessed directly via it's Project DOI: 10.21228/M8899W This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
| Study ID | ST002532 |
| Study Title | Nontargeted metabolomics analysis on kidney tissue treated with cisplatin |
| Study Type | Nontargeted metabolomics analysis |
| Study Summary | Cisplatin-induced acute kidney injury (AKI) is a severe clinical complication with no satisfactory therapies in the clinic. Tumor necrosis factor receptor (TNFR)-associated factor 1 (TRAF1) plays an important role in both inflammation and metabolism. However, the role of TRAF1 in cisplatin-induced AKI needs to be evaluated. In this study, TRAF1 expression was decreased in cisplatin-treated mice and mouse proximal tubular cells (mPTCs), suggesting a potential role of TRAF1 in cisplatin-associated kidney injury. Thus, TRAF1 plasmids were introduced into male C57BL/6J mice by a tail vein high-pressure injection method to overexpress TRAF1. Then, cisplatin was administrated by a single intraperitoneal (i.p.) injection (20 mg/kg). Mice were sacrificed 72 h after cisplatin administration. Serum and kidney tissues were collected for further analysis. In vitro, mPTCs were transfected with TRAF1 plasmids before treatment with cisplatin (5 µg/mL) for 24 h. Western blotting, Masson’s trichrome and hematoxylin-eosin (HE) staining and tandem mass spectrometry (LC‒MS/MS) analysis were employed to evaluate kidney injury. Following cisplatin treatment, we observed a marked downregulation of TRAF1 in mouse kidneys and mPTCs treated with cisplatin. In mice, TRAF1 overexpression attenuated cisplatin-induced AKI, as evidenced by decreased levels of serum creatinine (Scr) and blood urea nitrogen (BUN). Moreover, TRAF1 delivery obviously ameliorated cisplatin-induced renal tubular injury, as shown by the improved histological damage and blocked upregulation of NGAL and KIM-1. Moreover, the NF-κB activation and inflammatory cytokine production enhanced by cisplatin were significantly blunted by TRAF1. In line with the attenuated inflammatory response, the increased number of apoptotic cells (TUNEL staining) and enhanced expression of BAX and cleaved Caspase-3 were markedly decreased by TRAF1 overexpression. In vitro, TRAF1 also attenuated renal tubular cell inflammation and apoptosis induced by cisplatin. In addition, disordered cellular metabolism, which was reported as an important pathogenic factor of AKI, was examined by metabolomics analysis. Interestingly, a significant correction of the metabolic disturbance, including perturbations in energy generation and lipid and amino acid metabolism, was observed in the kidneys of cisplatin-treated mice. In conclusion, TRAF1 overexpression significantly attenuated cisplatin-induced nephrotoxicity, possibly by correcting the impaired metabolism, inhibiting inflammation, and blocking apoptosis in renal tubular cells. |
| Institute | Children's Hospital of Nanjing Medical University |
| Last Name | xiaolu |
| First Name | zhang |
| Address | 72 Guangzhou Road |
| zxiaolu0802@163.com | |
| Phone | 18351976523 |
| Submit Date | 2023-03-21 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-03-21 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001628 |
| Project DOI: | doi: 10.21228/M8899W |
| Project Title: | Nontargeted metabolomics studies on kidney tissue treated with cisplatin |
| Project Type: | LC?MS |
| Project Summary: | Nontargeted metabolomics analysis was carried out using the kidney tissues of mice killed at Day 3 after cisplatin treatment. |
| Institute: | Children's Hospital of Nanjing Medical University |
| Last Name: | xiaolu |
| First Name: | zhang |
| Address: | 72 Guangzhou Road |
| Email: | zxiaolu0802@163.com |
| Phone: | 18351976523 |
Subject:
| Subject ID: | SU002632 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Genotype | Strain |
|---|---|---|---|
| SA254732 | T-A14 | cisplatin+TRAF1 | WT |
| SA254733 | T-A13 | cisplatin+TRAF1 | WT |
| SA254734 | T-A11 | cisplatin+TRAF1 | WT |
| SA254735 | T-A15 | cisplatin+TRAF1 | WT |
| SA254736 | T-A12 | cisplatin+TRAF1 | WT |
| SA254737 | T-A3 | control | WT |
| SA254738 | T-A4 | control | WT |
| SA254739 | T-A5 | control | WT |
| SA254740 | T-A1 | control | WT |
| SA254741 | T-A2 | control | WT |
| SA254742 | T-A8 | vehicle+cisplatin | WT |
| SA254743 | T-A6 | vehicle+cisplatin | WT |
| SA254744 | T-A7 | vehicle+cisplatin | WT |
| SA254745 | T-A9 | vehicle+cisplatin | WT |
| SA254746 | T-A10 | vehicle+cisplatin | WT |
| Showing results 1 to 15 of 15 |
Collection:
| Collection ID: | CO002625 |
| Collection Summary: | 8 weeks old C57BL/6 male mice were randomly divided into three groups: control group (control; n=8), cisplatin-induced AKI group (cisplatin; n=8), and cisplatin plus TRAF1 plasmid injection group (cisplatin + TRAF1; n=8). Both cisplatin and cisplatin + TRAF1 groups were given 20 mg/kg cisplatin intraperitoneal (i.p.) injection, while the control group received the same amount of saline i.p. injection. The mice in cisplatin + TRAF1 group were pretreated with TRAF1 performed by a hydrodynamic-based gene delivery approach 36 h before cisplatin administration. Mice in the three groups were sacrificed after cisplatin treatment for 72 h. The kidney tissues were cut horizontally and collected. |
| Collection Protocol Filename: | protocol.pdf |
| Sample Type: | Kidney |
Treatment:
| Treatment ID: | TR002644 |
| Treatment Summary: | 8 weeks old C57BL/6 male mice were randomly divided into three groups: control group (control; n=8), cisplatin-induced AKI group (cisplatin; n=8), and cisplatin plus TRAF1 plasmid injection group (cisplatin + TRAF1; n=8). Both cisplatin and cisplatin + TRAF1 groups were given 20 mg/kg cisplatin intraperitoneal (i.p.) injection, while the control group received the same amount of saline i.p. injection. The mice in cisplatin + TRAF1 group were pretreated with TRAF1 performed by a hydrodynamic-based gene delivery approach 36 h before cisplatin administration. Mice in the three groups were sacrificed after cisplatin treatment for 72 h. The kidney tissues were cut horizontally and collected. |
| Treatment Protocol Filename: | protocol.pdf |
Sample Preparation:
| Sampleprep ID: | SP002638 |
| Sampleprep Summary: | 1. Take all samples into a 2 mL EP tube, add 2 steel balls, put them into the tissue grinder, grind at 50 Hz for 60 s, and then homogenize the samples; 2. Accurately weigh 100 mg (±1%) of the homogenized sample into a 2 mL EP tube, add accurately 0.6mL of methanol (including internal standard), and vortex for 30 s 3. Add 2 steel balls, put them into the tissue grinder, and grind for 60 s at 50 Hz; 4. Centrifuge at 4 ℃ for 10 min at 12000 rpm, and the supernatant was filtered through 0.22 µm membrane to obtain the prepared samples for LC-MS; 5. Take 20 µL from each sample to the quality control (QC) samples; (These QC samples were used to monitor deviations of the analytical results from these pool mixtures and compare them to the errors caused by the analytical instrument itself) 6. Use the rest of the samples for LC-MS detection. |
| Sampleprep Protocol Filename: | protocol.pdf |
Combined analysis:
| Analysis ID | AN004166 | AN004167 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Thermo Ultimate 3000 | Thermo Ultimate 3000 |
| Column | Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.8um) | Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.8um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Focus | Thermo Q Exactive Focus |
| Ion Mode | NEGATIVE | POSITIVE |
| Units | peak area | peak area |
Chromatography:
| Chromatography ID: | CH003085 |
| Methods Filename: | protocol.pdf |
| Instrument Name: | Thermo Ultimate 3000 |
| Column Name: | Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.8um) |
| Column Temperature: | 40 |
| Flow Gradient: | 0~1 min, 2% B/D; 1~9 min, 2%~50% B/D; 9~12 min, 50%~98% B/D; 12~13.5 min, 98% B/D; 13.5~14 min, 98%~2% B/D; 14~20 min, 2% D-positive model (14~17 min, 2% B-negative model) |
| Flow Rate: | 0.25 mL/min |
| Solvent A: | 0.1% formic acid in water and 0.1% formic acid in acetonitrile |
| Solvent B: | 5 mM ammonium formate in water and acetonitrile |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003913 |
| Analysis ID: | AN004166 |
| Instrument Name: | Thermo Q Exactive Focus |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | - |
| Ion Mode: | NEGATIVE |
| Analysis Protocol File: | protocol.pdf |
| MS ID: | MS003914 |
| Analysis ID: | AN004167 |
| Instrument Name: | Thermo Q Exactive Focus |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | - |
| Ion Mode: | POSITIVE |
| Analysis Protocol File: | protocol.pdf |
