Dataset Information

The impact of exercise on skeletal muscle proteome of prediabetic subjects analyzed with data independent mass spectrometry.

ABSTRACT: Skeletal muscle is a primary tissue responsible for both the exercise capacity and insulin-induced glucose uptake. Exercise-evoked proteomic alternations in muscle of subjects with prediabetes are of great importance for the elucidation of the connections between insulin resistance and physical exercise. To envision best aviable approach, we compared various ion-chromatogram libraries assembled with the use of off-line high-pH fractionation (HpH), gas-phase fractionation (GPF) and library-less DirectDIATM approach in LC/MS/HRMS DIA proteomic analysis of muscle from normoglycemic (NGT) and prediabetic (IGT) subjects after 3 months of supervised, mixed-mode exercise. Although HpH approach to library generation resulted in best proteome coverage, quantitative and qualitative analysis of HeLa standards, mixed samples and whole study set revealed, that GPF-fractionated hybrid DDA/DIA-acquired libraries yielded best overall balance between the speed of preparation, data collection, data analysis and identification in real-life clinical samples.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Skeletal Muscle Fiber, Vastus Lateralis

DISEASE(S): Prediabetes Syndrome

SUBMITTER: Piotr Zabielski

LAB HEAD: Piotr Zabielski

PROVIDER: PXD055536 | Pride | 2025-07-22

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	20240902_174109_BESD_NGT_vs_IGT_GPF_COMBINED_Report.setup.txt	Txt
	20240902_174109_BESD_NGT_vs_IGT_GPF_COMBINED_Report.tsv	Tabular
	20240902_180101_BESD_NGT_vs_IGT_HPH_COMBINED_Report.setup.txt	Txt
	20240902_180101_BESD_NGT_vs_IGT_HPH_COMBINED_Report.tsv	Tabular
	20240902_180943_BESD_NGT_vs_IGT_DirectDIA_COMBINED_Report.setup.txt	Txt

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Project description:Relatively brief bouts of exercise alter gene expression in peripheral blood mononuclear cells (PBMCs), but whether or not exercise changes gene expression in circulating neutrophils (whose numbers, like PBMCs, increase) is not known. We hypothesized that exercise would activate neutrophil genes involved in apoptosis, inflammation, and cell growth and repair, since these functions in leukocytes are known to be influenced by exercise. Blood was sampled before and immediately after 30-min of constant, heavy (about 80% peak O2 uptake) cycle-ergometer exercise in 12 healthy men (19-29 yr old) of average fitness. Neutrophils were isolated using density gradients; RNA was hybridized to Affymetrix U133+2 Genechip arrays. Using FDR<0.05 with 95% confidence a total of 526 genes were differentially expressed between before and after exercise. 316 genes had higher expression after exercise. The Jak/STAT pathway, known to inhibit apoptosis, was significantly activated (EASE score, p<0.005), but 14 genes were altered in a way likely to accelerate apoptosis as well. Similarly, both proinflammatory (e.g., IL32, TNFSF8 and CCR5) and anti-inflammatory (e.g., ANXA1) were affected. Growth and repair genes like AREG and FGF2 receptor genes (involved in angiogenesis) were also activated. Finally, a number of neutrophil genes known to be involved in pathological conditions like asthma and arthritis were altered by exercise, suggesting novel links between physical activity and disease or its prevention. In summary, brief heavy exercise leads to a previously unknown substantial and significant alteration in neutrophil gene expression. Experiment Overall Design: Twelve healthy men (19-29 yr old) participated in this study. A baseline blood sample was taken before the onset of exercise and immediately after 30-min exercise bout. Neutrophils were isolated using OptiPrep Density Gradient Medium (SIGMA). Total RNA was extracted using TRIzol. cRNA was hybridized onto Affymetrix U133+2 arrays (total of 24 chips).

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The impact of exercise on skeletal muscle proteome of prediabetic subjects analyzed with data independent mass spectrometry.

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