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Transforming growth factor-β1-induced podocyte injury is associated with increased microRNA-155 expression, enhanced
Xintong Zheng1, Qiuhong Zhong2, Xu Lin1
1Department of Nephrology, The Affiliated Hospital of Youjiang Medical University for Nationalities, Baise, Guangxi 533000, P.R. China.
Abstract:
MicroRNA-155 (miR-155) is associated with various diseases. However, the potential role of miR-155 in early glomerular disease (EGD) remains elusive. In the present study, the clinical significance of urinary miR-155 expression was explored in patients with EGD using receiver operating characteristic curve analysis. Conditionally immortalized mouse podocytes were cultured in vitro and treated with transforming growth factor-β1 (TGF-β1) at different concentrations and durations. The gene expression levels of mRNAs and miR-155 were detected using reverse transcription-quantitative PCR. Synaptopodin, CD2-associated protein (CD2AP), p38, and extracellular signal-regulated kinase (Erk) 1/2 expressions were detected using western blotting. Cell supernatants were collected for assaying tumor necrosis factor (TNF)-α and interleukin (IL)-6 concentrations using enzyme-linked immunosorbent assay. The Pearson correlation analysis was used to analyze the correlation between miR-155 levels and TNF-α or IL-6. It was found that miR-155 levels in urine have high sensitivity and specificity in the diagnosis of EGD. Time- and dose-dependent TGF-β1 treatments downregulated synaptopodin and CD2AP expression levels, and activated the p38 and Erk 1/2 pathway. However, these effects were attenuated by p38 and Erk 1/2 phosphorylation inhibitors. Additionally, TNF-α and IL-6 secretions were elevated, and their concentrations were positively correlated with the expression of miR-155 during podocyte injury. Thus, the present study indicated that miR-155 is a potential biomarker for the diagnosis of EGD, and its expression is associated with the release of pro-inflammatory cytokines and activation of mitogen-activated protein kinase (MAPK) pathway in TGF-β1-induced podocyte injury. The present study suggests that the TGF-β1/miR-155/MAPK axis is a novel target in the mechanism of EGD.
Insights
Urinary microRNA-155 (miR-155) shows high accuracy for diagnosing early glomerular disease (EGD). Its levels correlate with inflammatory cytokines and MAPK pathway activation in podocyte injury.
Area of Science:
- Nephrology
- Molecular Biology
- Biomarker Discovery
Background:
- MicroRNA-155 (miR-155) is implicated in various diseases.
- The role of miR-155 in early glomerular disease (EGD) is not well understood.
- Identifying reliable biomarkers for EGD is crucial for early diagnosis and intervention.
Purpose of the Study:
- To investigate the clinical significance of urinary miR-155 as a diagnostic biomarker for EGD.
- To explore the molecular mechanisms involving miR-155 in transforming growth factor-β1 (TGF-β1)-induced podocyte injury.
- To examine the relationship between miR-155, pro-inflammatory cytokines, and the mitogen-activated protein kinase (MAPK) pathway.
Main Methods:
- Urinary miR-155 levels were analyzed in EGD patients using receiver operating characteristic (ROC) curve analysis.
- In vitro studies cultured mouse podocytes treated with TGF-β1, assessing gene and protein expression (mRNA, miR-155, synaptopodin, CD2AP, p38, Erk1/2) via RT-qPCR and Western blotting.
- Enzyme-linked immunosorbent assay (ELISA) measured tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) concentrations; Pearson correlation analyzed miR-155 and cytokine levels.
Main Results:
- Urinary miR-155 demonstrated high sensitivity and specificity for EGD diagnosis.
- TGF-β1 treatment downregulated synaptopodin and CD2AP, activated p38 and Erk1/2 pathways, and increased TNF-α and IL-6 secretion.
- Elevated miR-155 levels positively correlated with increased TNF-α and IL-6 concentrations during podocyte injury.
Conclusions:
- Urinary miR-155 serves as a potential diagnostic biomarker for EGD.
- miR-155 expression is linked to pro-inflammatory cytokine release and MAPK pathway activation in TGF-β1-induced podocyte injury.
- The TGF-β1/miR-155/MAPK axis represents a novel therapeutic target for EGD.
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