MetaboLights

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data after the database search is uploaded to the Majorbio cloud platform (https://cloud.majorbio.com) for data analysis. Metabolic features detected at least 80% in any set of samples were retained. After filtering, minimum metabolite values were imputed for specific samples in which the metabolite levels fell below the lower limit of quantitation and each Metabolic features were normalized by sum. In order to reduce the errors caused by sample preparation and instrument instability, the response intensity of the sample mass spectrum peaks was normalized by the sum normalization method, and the normalized data matrix was obtained. At the same time, variables with relative standard deviation (RSD) > 30% of QC samples were removed, and log10 logarithmization was performed to obtain the final data matrix for subsequent analysis.MetaboLightsPublicLiquid Chromatography MS - negative - reverse phaseLiquid Chromatography MS - positive - reverse phaseA total of 2 μL of sample was separated using a Thermo Scientific Vanquish Horizon UHPLC System and a ACQUITY UPLC HSS T3 (1.8 µm, 2.1 mm x 100 mm; Waters) column. The mobile phases consisted of 0.1% formic acid in water:acetonitrile (95:5, v/v) (solvent A) and 0.1% formic acid in acetonitrile:isopropanol:water (47.5:47.5:5, v/v) (solvent B). The solvent gradient changed according to the following conditions: from 0.0 to 0.1 min, from 0% to 5% B; from 0.1 to 2.0 min, from 5% to 25% B; from 2.0 to 9.0 min, from 25% to 100% B; from 9.0 to 13.0 min, from 100% to 100% B; from 13.0 to 13.1 min, from 100% to 0% B ; from 13.1 to 16.0 min, from 0% to 0% B for equilibrating the systems. The sample injection volume was 2 µL and the flow rate was set to 0.4 mL/min. The column temperature was maintained at 40 °C. During the period of analysis, all these samples were stored at 4 °C.Multi-omics analysis revealed spermidine contributes to ovarian function improvement.chengweng JiSichuan agricultural universityblood serummass spectrometryBriefly, 200 μL liquid sample were extracted using a 400 µL methanol:acetonitrile (1:1, v/v) solution. The mixture then sonicated at 40 kHz for 30 min at 5 °C. The samples were placed at -20 °C for 30 min to precipitate proteins. After centrifugation at 13,000 x g at 4 °C for 15 min, the supernatant were carefully transferred to new microtubes and evaporated to dryness under a gentle stream of nitrogen. For UHPLC-MS/MS anlaysis, the samples were reconstituted in 100 µL loading solution of acetonitrile:water (1:1, v/v) by brief sonication in a 5 °C water bath. Extracted metabolites were spun for 15 min at 13,000 x g at 4 °C on a bench-top centrifuge and cleared supernatant were transferred to sample vials for LC-MS/MS analysis.Mus musculushttps://www.ebi.ac.uk/metabolights/MTBLS11115Shuo Li. Sichuan Agricultural University, Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china. Sichuan Agricultural University, Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china. 2023202044@stu.sicau.edu.cn.Lu Lu. Sichuan Agricultural University, Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china. Sichuan Agricultural University, Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china. lu.lu@sicau.edu.cn.Xiaoguang An. Sichuan Agricultural University, Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china. Sichuan Agricultural University, Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china. anxiaoguang@stu.sicau.edu.cn.Kang Bo. Sichuan Agricultural University, Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china. Sichuan Agricultural University, Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china. bokang@sicau.edu.cn.Mingzhou Li. Sichuan Agricultural University, Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china. Sichuan Agricultural University, Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china. mingzhou.li@sicau.edu.cn.Chengweng Ji. Sichuan Agricultural University，Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china. jichengweng@stu.sicau.edu.cn. Sichuan Agricultural University，Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china.Xin Wang. Sichuan Agricultural University，Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china. Sichuan Agricultural University，Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china. xinwang@stu.sicau.edu.cn.Yuxin Qi. Sichuan Agricultural University，Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china. Sichuan Agricultural University，Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china. qiyuxin@stu.sicau.edu.cn.Dongmei Jiang. Sichuan Agricultural University，Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china. Sichuan Agricultural University，Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china. jiangdm@sicau.edu.cn.Weijie Zhang. Sichuan Agricultural University，Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china. Sichuan Agricultural University，Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china. 1550214884@qq.com.Weikang Ling. Sichuan Agricultural University, Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china. Sichuan Agricultural University, Huimin Street, Wenjiang District, Chengdu City, Sichuan Province，china. lingweikang@stu.sicau.edu.cn.The data after the database search is uploaded to the Majorbio cloud platform (https://cloud.majorbio.com) for data analysis. Metabolic features detected at least 80% in any set of samples were retained. After filtering, minimum metabolite values were imputed for specific samples in which the metabolite levels fell below the lower limit of quantitation and each Metabolic features were normalized by sum. In order to reduce the errors caused by sample preparation and instrument instability, the response intensity of the sample mass spectrum peaks was normalized by the sum normalization method, and the normalized data matrix was obtained. At the same time, variables with relative standard deviation (RSD) > 30% of QC samples were removed, and log10 logarithmization was performed to obtain the final data matrix for subsequent analysis.WGE dose1097423037@qq.comThe experimental animals included 189 6-week-old C57BL/6 female mice and 35 male mice purchased from Chengdu Dossy Experimental Animals Co., Ltd (Chengdu, China). Female mice were divided into 4 groups: a 0 g/kg WGE group (control), 50 g/kg WGE group (WGE50), 100 g/kg WGE group (WGE100) and 150 g/kg WGE group (WGE150). These groups corresponded to daily spermidine consumption of 0 mg/kg, 250 mg/kg, 500 mg/kg and 750 mg/kg body weight. Each group comprised 7 cages of mice, with 5 mice per cage. All mice were pre-fed 1 week in advance. The feed was then changed to WGE feed, and the mice were fed for 12 weeks. Food and water were provided ad libitum. Before euthanasia, blood was collected from the hearts of anesthetized mice with a 1 mL syringe. Subsequently, the blood was centrifuged at 4 °C for 10 min at 3000 rpm to separate serum samples for LC-MS analysis.Metabolomicsspermidinehigh-performance liquid chromatography-mass spectrometryuntargeted metabolitesMultiomicsspermidinehigh-performance liquid chromatography-mass spectrometryuntargeted metabolitesMultiomicsThe mass spectrometric data was collected using a Thermo UHPLC-Q Exactive Mass Spectrometer equipped with an electrospray ionization (ESI) source operating in either positive or negative ion mode. The optimal conditions were set as followed: heater temperature, 400 °C; 6Capillary temperature, 320 °C; sheath gas flow rate, 40 arb; Aux gas flow rate, 10 arb; ion-spray voltage floating (ISVF), -2800 V in negative mode and +3500 V in positive mode, respectively; Normalized collision energy, 20-40-60 V rolling for MS/MS. Full MS resolution was 70000, and MS/MS resolution was 17500. Data acquisition was performed with the Data Dependent Acquisition (DDA) mode. The detection was carried out over a mass range of 70-1050 m/z.falseMulti-omics analysis revealed spermidine contributes to ovarian function improvementSpermidine is involved in extensive cellular processes, including mammalian oocyte development. Wheat germ is a natural source of polyamines and contains high concentrations of spermidine. No studies have assessed the effects and mechanisms of action of wheat germ spermidine in regulating mammalian ovarian functions. Herein, a dietary experiment was conducted in mice administered wheat germ extract (WGE) for 90 days. Dietary WGE significantly increased spermidine absorption and affected the metabolism of the other polyamines (spermine and putrescine) in mouse intestines. Dietary intake of 50 g/kg WGE significantly increased the number of mouse pups per litter (p < 0.01), and the secreted levels of the sex hormones E2 and P4 (p < 0.05). Furthermore, compared with a normal diet, WGE feeding led to significantly greater ovarian reserves and fewer atretic follicles (p < 0.001). Finally, co-relationship analysis revealed gut microbes that significantly correlated with sex hormone levels. In summary, our results confirm that wheat germ spermidine plays a beneficial role in mouse ovarian function. Moreover, changes in the structure of the gut microbiota are likely to be crucial for improving ovarian function.2024-10-282024-09-18MTBLS11115