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Updated: Aug 7, 2026
Lipid Droplet Isolation for Quantitative Mass Spectrometry Analysis
Published on: April 17, 2017
Cytokine mRNA profile in lipoid nephrosis: evidence for increased IL-8 mRNA stability
Paul F Laflam1, Soichi Haraguchi, Eduardo H Garin
1Division of Nephrology, Department of Pediatrics, University of South Florida, Tampa 33606, USA.
Background/Aims:
Proteinuria in idiopathic minimal lesion nephrotic syndrome (IMLNS) is presumed to be due to the effect of circulating factors on glomerular permeability to plasma proteins. This study examines the expression of messenger ribonucleic acid (mRNA) for cytokines thought to mediate glomerular inflammation during different stages of the nephrotic syndrome.
Methods:
Messenger RNA expression and stability from peripheral blood mononuclear cells of IMLNS patients in relapse and in remission, and age matched normal controls were measured using a ribonuclease protection assay (RPA). The spontaneous and Interleukin 2 (IL-2) stimulated mRNA expression were studied.
Results:
Spontaneous mRNA expression for Interleukin 8 (IL-8) from IMLNS patients in relapse was significantly increased when compared to IMLNS patients in remission and normal controls (p < 0.05). After 14 h of IL-2 stimulation, mRNA IL-8 levels expressed by IMLNS PBMC patients in remission were not different from those observed in normal controls. However, after 5 days of PBMC incubation, a significant increase in mRNA for IL-8 in IMLNS patients compared to controls was found (p < 0.01). Stability assay demonstrated that IL-8 mRNA transcript from the nephrotic patients remained higher than those from controls and showed a significantly prolonged life t(1/2) (p = 0.02).
Conclusions:
IL-8 mRNA expression is increased in IMLNS patients in relapse. Moreover, stability studies show that IL-8 mRNA life t(1/2) is prolonged due to altered post-transcriptional regulation. This finding may explain the elevated serum IL-8 levels observed in these patients during relapse and may have pathogenic significance since IL-8 has been shown to induce proteinuria in the experimental animal.
Insights
Interleukin 8 (IL-8) messenger RNA (mRNA) expression is elevated in patients with idiopathic minimal lesion nephrotic syndrome (IMLNS) during relapse. This increase is linked to prolonged IL-8 mRNA stability, potentially explaining proteinuria in IMLNS.
Area of Science:
- Nephrology
- Immunology
- Molecular Biology
Background:
- Idiopathic minimal lesion nephrotic syndrome (IMLNS) proteinuria is linked to circulating factors affecting glomerular permeability.
- The role of cytokines in mediating glomerular inflammation in IMLNS requires further investigation.
Purpose of the Study:
- To examine messenger RNA (mRNA) expression of cytokines involved in glomerular inflammation during different stages of nephrotic syndrome.
- To investigate the expression and stability of Interleukin 8 (IL-8) mRNA in peripheral blood mononuclear cells (PBMCs) of IMLNS patients.
Main Methods:
- Messenger RNA (mRNA) expression and stability were measured using ribonuclease protection assay (RPA) in PBMCs from IMLNS patients (relapse and remission) and healthy controls.
- Spontaneous and Interleukin 2 (IL-2) stimulated mRNA expression of IL-8 was analyzed.
Main Results:
- Spontaneous IL-8 mRNA expression was significantly higher in IMLNS patients during relapse compared to remission and controls (p < 0.05).
- IL-8 mRNA levels in IMLNS patients showed a significant increase after prolonged incubation (5 days) with IL-2 compared to controls (p < 0.01).
- IL-8 mRNA transcripts from nephrotic patients exhibited prolonged half-life (t(1/2)), indicating altered post-transcriptional regulation (p = 0.02).
Conclusions:
- Increased IL-8 mRNA expression and prolonged IL-8 mRNA stability are observed in IMLNS patients during relapse.
- Altered post-transcriptional regulation contributes to elevated IL-8 mRNA levels and may explain increased serum IL-8 in relapse.
- Elevated IL-8 may play a pathogenic role in IMLNS, as IL-8 can induce proteinuria in experimental models.
