Summary of Study ST002298

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 PR001472. The data can be accessed directly via it's Project DOI: 10.21228/M8FT53 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.

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Study ID	ST002298
Study Title	NAD(P) deficiency plays an important role in the restraint-stress-induced depression in the rat model
Study Summary	The metabolic dysfunction or irreversible metabolic changes from stress may cause body vulnerability, potentially leading to the onset of psychiatric and non-psychiatric illnesses. Nevertheless, little is known about the biochemical events that cause depression due to stress. Our study employed open field test, plasma adrenocorticotropic hormone (ACTH) and corticosterone determination, serum biochemical analysis, quantitative PCR, immunoblotting, enzyme activity assay, and NMR-based metabolomics to analyze and identify the biochemical variations of body fluids (serum and urine) and tissues (brain, kidney, liver, lung, and spleen) in an acute restraint stress-induced rat model of depression. Our data suggested that the post-stress effects on biochemical alterations involved different biochemical pathways, including regulating the NAD(P) pool, glucose homeostasis, biosynthesis and degradation of heme, and uric acid production and metabolism. The urinary excretion of nicotinate and nicotinamide N-oxide increased significantly. Thus, we conclude that the depletion of NAD(P) precursors may occur in response to restraint stress. Our results show a close association between NAD(P) deficiency and post-stress metabolic dysfunction, which would provide a ground for developing recovery-promoting micronutrients in treating depression.
Institute	Anhui Science and Technology University
Last Name	Li
First Name	Jinquan
Address	No. 9, Donghua Road, Fengyang, Anhui Province, 233100, China
Email	lijinquan@ahstu.edu.cn
Phone	86 133 2875 1890
Submit Date	2022-07-30
Raw Data Available	Yes
Raw Data File Type(s)	fid
Analysis Type Detail	NMR
Release Date	2023-08-15
Release Version	1

Select appropriate tab below to view additional metadata details:

Project:

Project ID:	PR001472
Project DOI:	doi: 10.21228/M8FT53
Project Title:	NAD(P) deficiency plays an important role in the restraint-stress-induced depression in the rat model
Project Summary:	The metabolic dysfunction or irreversible metabolic changes from stress may cause body vulnerability, potentially leading to the onset of psychiatric and non-psychiatric illnesses. Nevertheless, little is known about the biochemical events that cause depression due to stress. Our study employed open field test, plasma adrenocorticotropic hormone (ACTH) and corticosterone determination, serum biochemical analysis, quantitative PCR, immunoblotting, enzyme activity assay, and NMR-based metabolomics to analyze and identify the biochemical variations of body fluids (serum and urine) and tissues (brain, kidney, liver, lung, and spleen) in an acute restraint stress-induced rat model of depression. Our data suggested that the post-stress effects on biochemical alterations involved different biochemical pathways, including regulating the NAD(P) pool, glucose homeostasis, biosynthesis and degradation of heme, and uric acid production and metabolism. The urinary excretion of nicotinate and nicotinamide N-oxide increased significantly. Thus, we conclude that the depletion of NAD(P) precursors may occur in response to restraint stress. Our results show a close association between NAD(P) deficiency and post-stress metabolic dysfunction, which would provide a ground for developing recovery-promoting micronutrients in treating depression.
Institute:	Anhui Science and Technology University
Last Name:	Li
First Name:	Jinquan
Address:	No. 9, Donghua Road, Fengyang, Anhui Province, 233100, China
Email:	lijinquan@ahstu.edu.cn
Phone:	86 133 2875 1890

Subject:

Subject ID:	SU002384
Subject Type:	Mammal
Subject Species:	Rattus norvegicus
Taxonomy ID:	10116

Factors:

Subject type: Mammal; Subject species: Rattus norvegicus (Factor headings shown in green)

mb_sample_id	local_sample_id	Experimental factor
SA220958	urine_44311	24 h post-stress
SA220959	urine_44312	24 h post-stress
SA220960	urine_44313	24 h post-stress
SA220961	urine_44310	24 h post-stress
SA220962	urine_44309	24 h post-stress
SA220963	urine_44314	24 h post-stress
SA220964	urine_44308	24 h post-stress
SA220965	liver_44084	48 h post-stress
SA220966	liver_44079	48 h post-stress
SA220967	spleen_44101	48 h post-stress
SA220968	spleen_44096	48 h post-stress
SA220969	lung_44107	48 h post-stress
SA220970	lung_44102	48 h post-stress
SA220971	liver_44074	48 h post-stress
SA220972	lung_44097	48 h post-stress
SA220973	liver_44089	48 h post-stress
SA220974	liver_44094	48 h post-stress
SA220975	kidney_44085	48 h post-stress
SA220976	kidney_44080	48 h post-stress
SA220977	kidney_44090	48 h post-stress
SA220978	kidney_44095	48 h post-stress
SA220979	kidney_44105	48 h post-stress
SA220980	kidney_44100	48 h post-stress
SA220981	kidney_44075	48 h post-stress
SA220982	spleen_44106	48 h post-stress
SA220983	liver_44104	48 h post-stress
SA220984	liver_44099	48 h post-stress
SA220985	lung_44092	48 h post-stress
SA220986	lung_44087	48 h post-stress
SA220987	lung_44077	48 h post-stress
SA220988	lung_44082	48 h post-stress
SA220989	spleen_44076	48 h post-stress
SA220990	urine_44318	48 h post-stress
SA220991	serum_44010	48 h post-stress
SA220992	serum_44009	48 h post-stress
SA220993	serum_44011	48 h post-stress
SA220994	serum_44012	48 h post-stress
SA220995	serum_44013	48 h post-stress
SA220996	serum_44008	48 h post-stress
SA220997	serum_44007	48 h post-stress
SA220998	brain_44108	48 h post-stress
SA220999	brain_44103	48 h post-stress
SA221000	brain_44098	48 h post-stress
SA221001	brain_44093	48 h post-stress
SA221002	urine_44321	48 h post-stress
SA221003	brain_44088	48 h post-stress
SA221004	urine_44316	48 h post-stress
SA221005	urine_44317	48 h post-stress
SA221006	urine_44319	48 h post-stress
SA221007	urine_44320	48 h post-stress
SA221008	brain_44083	48 h post-stress
SA221009	urine_44315	48 h post-stress
SA221010	spleen_44091	48 h post-stress
SA221011	spleen_44081	48 h post-stress
SA221012	spleen_44086	48 h post-stress
SA221013	brain_44078	48 h post-stress
SA221014	lung_44052	non-stress
SA221015	lung_44057	non-stress
SA221016	lung_44047	non-stress
SA221017	brain_44053	non-stress
SA221018	brain_44063	non-stress
SA221019	brain_44068	non-stress
SA221020	brain_44073	non-stress
SA221021	brain_44058	non-stress
SA221022	brain_44048	non-stress
SA221023	lung_44067	non-stress
SA221024	lung_44072	non-stress
SA221025	lung_44062	non-stress
SA221026	kidney_44060	non-stress
SA221027	urine_44305	non-stress
SA221028	urine_44304	non-stress
SA221029	urine_44306	non-stress
SA221030	urine_44307	non-stress
SA221031	liver_44049	non-stress
SA221032	liver_44044	non-stress
SA221033	urine_44303	non-stress
SA221034	urine_44302	non-stress
SA221035	serum_44003	non-stress
SA221036	serum_44002	non-stress
SA221037	serum_44004	non-stress
SA221038	serum_44005	non-stress
SA221039	serum_44006	non-stress
SA221040	liver_44054	non-stress
SA221041	liver_44059	non-stress
SA221042	spleen_44046	non-stress
SA221043	kidney_44070	non-stress
SA221044	spleen_44051	non-stress
SA221045	spleen_44056	non-stress
SA221046	spleen_44066	non-stress
SA221047	spleen_44061	non-stress
SA221048	kidney_44065	non-stress
SA221049	serum_44001	non-stress
SA221050	liver_44069	non-stress
SA221051	liver_44064	non-stress
SA221052	kidney_44045	non-stress
SA221053	kidney_44050	non-stress
SA221054	kidney_44055	non-stress
SA221055	spleen_44071	non-stress

Showing results 1 to 98 of 98

Showing results 1 to 98 of 98

Collection:

Collection ID:	CO002377
Collection Summary:	Individual urine samples were collected in ice-cooled vessels containing 1% sodium azide (0.1 ml) for 2 h using a metabolic cage at 0, 24, and 48 h post-stress, respectively, and immediately frozen at -80°C. Animals were sacrificed by exsanguination under isoflurane anesthesia at 48 h post-stress. The blood sample was divided into two aliquots, one serum for biochemical analysis and the other heparinized plasma for NMR spectroscopic analysis. After weighing, brain, kidney, liver, lung, and spleen tissue were excised in duplicate: one being fixed in 10% formalin for histopathological examination, the other immediately snap-frozen in liquid nitrogen for tissue extraction. These samples were stored at -80°C until used.
Collection Protocol Filename:	Protocols.pdf
Sample Type:	serum, urine, brain, kidney, liver, lung, spleen

Treatment:

Treatment ID:	TR002396
Treatment Summary:	According to the National Institutes of Health Guide for the Care and Use of Laboratory Animals, all animals involved in this study were cared for, and protocols were reviewed and approved by the Anhui Laboratory Animal Care Committee. The specific pathogen-free (SPF) seven-week-old male Sprague Dawley (SD) rats (weight 233 ± 5 g) were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd and used in this study. The environmental conditions were set at 21-26°C with a relative humidity of 50 ± 10% and a 12/12 h light/dark cycle. Food and tap water were provided ad libitum, and body weights were recorded daily. After one week of acclimatization, rats were randomly assigned to the groups of non-stressed control (n = 6) or the stressed (n = 7). For restraint stress, rats were individually placed in a ventilated plastic tube restrainer for 120 minutes, using a previously modified method. According to the general protocol, control rats were left undistributed in a home cage and allowed to contact each other without food and water.

Treatment ID:

TR002396

Treatment Summary:

According to the National Institutes of Health Guide for the Care and Use of Laboratory Animals, all animals involved in this study were cared for, and protocols were reviewed and approved by the Anhui Laboratory Animal Care Committee. The specific pathogen-free (SPF) seven-week-old male Sprague Dawley (SD) rats (weight 233 ± 5 g) were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd and used in this study. The environmental conditions were set at 21-26°C with a relative humidity of 50 ± 10% and a 12/12 h light/dark cycle. Food and tap water were provided ad libitum, and body weights were recorded daily. After one week of acclimatization, rats were randomly assigned to the groups of non-stressed control (n = 6) or the stressed (n = 7). For restraint stress, rats were individually placed in a ventilated plastic tube restrainer for 120 minutes, using a previously modified method. According to the general protocol, control rats were left undistributed in a home cage and allowed to contact each other without food and water.

Sample Preparation:

Sampleprep ID:	SP002390
Sampleprep Summary:	Samples of plasma (255 μl) were mixed with 255 μl of phosphate D2O buffer solution (NaH2PO4 and K2HPO4, 60 mM, pH 7.4). After centrifugation at 10000 × g at 4°C for 10 min to remove the precipitates, the supernatants were transferred to 5 mm NMR tubes and analyzed by NMR. Samples of urine (455 μl) were mixed with 55 μl of D2O buffer solution (NaH2PO4 and K2HPO4, 1.5 M, including 0.1% TSP (sodium 3-(trimethylsilyl) propionate-2,2,3,3-d4), pH 7.4) to minimize any gross variation in the pH of the urine samples. The mixture was left to stand for 10 min and centrifuged at 10000 × g at 4°C for 10 min to remove the precipitates. The supernatants were transferred to 5 mm NMR tubes and analyzed by NMR. The polar metabolites in the rat tissue were extracted according to the protocol established in our previous work. In brief, pre-weighed brain, kidney, liver, lung, or spleen samples (100 mg) were homogenized in 400 μl of CH3OH and 85 μl of H2O at 4°C. The homogenates were transferred into a 2.5-ml tube, combined with 400 μl of CHCl3 and 200 μl of H2O, and then kept in a vortex for 60 s. After 10-min partitioning on ice, the samples were centrifuged for 5 min (10000 × g, 4°C). The upper supernatants were transferred into 1.5-ml tubes and lyophilized to remove CH3OH and H2O. The extracts were reconstituted in 0.5 ml D2O containing 1 mM TSP, then transferred into 5-mm NMR tubes and analyzed by NMR spectroscopy.

Sampleprep ID:

SP002390

Sampleprep Summary:

Samples of plasma (255 μl) were mixed with 255 μl of phosphate D2O buffer solution (NaH2PO4 and K2HPO4, 60 mM, pH 7.4). After centrifugation at 10000 × g at 4°C for 10 min to remove the precipitates, the supernatants were transferred to 5 mm NMR tubes and analyzed by NMR. Samples of urine (455 μl) were mixed with 55 μl of D2O buffer solution (NaH2PO4 and K2HPO4, 1.5 M, including 0.1% TSP (sodium 3-(trimethylsilyl) propionate-2,2,3,3-d4), pH 7.4) to minimize any gross variation in the pH of the urine samples. The mixture was left to stand for 10 min and centrifuged at 10000 × g at 4°C for 10 min to remove the precipitates. The supernatants were transferred to 5 mm NMR tubes and analyzed by NMR. The polar metabolites in the rat tissue were extracted according to the protocol established in our previous work. In brief, pre-weighed brain, kidney, liver, lung, or spleen samples (100 mg) were homogenized in 400 μl of CH3OH and 85 μl of H2O at 4°C. The homogenates were transferred into a 2.5-ml tube, combined with 400 μl of CHCl3 and 200 μl of H2O, and then kept in a vortex for 60 s. After 10-min partitioning on ice, the samples were centrifuged for 5 min (10000 × g, 4°C). The upper supernatants were transferred into 1.5-ml tubes and lyophilized to remove CH3OH and H2O. The extracts were reconstituted in 0.5 ml D2O containing 1 mM TSP, then transferred into 5-mm NMR tubes and analyzed by NMR spectroscopy.

Analysis:

Analysis ID:	AN003754
Analysis Type:	NMR
Results File:	ST002298_AN003754_Results.txt
Units:	Peak area

NMR:

NMR ID:	NM000253
Analysis ID:	AN003754
Instrument Name:	Varian 500 MHz spectrometer/Bruker-AV600 spectrometer
Instrument Type:	FT-NMR
NMR Experiment Type:	1D-1H
Spectrometer Frequency:	500 MHz/600 MHz