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Background <p>Long-term peritoneal dialysis (PD) induces persistent inflammation and fibrosis of the peritoneal membrane (PM), altering the peritoneal solute transport rate (PSTR). We previously found elevated miR-503-5p levels in exosomes from PD effluent (PDE) in patients with high PSTR. As PSTR is strongly influenced by intraperitoneal inflammation, this study aimed to explore the relationship between miR-503-5p, inflammation, and peritoneal fibrosis and the underlying mechanisms.</p> Methods <p>Bioinformatics analyses were initially performed to identify the miR-503-5p target genes and determine the research subject. Real-time polymerase chain reaction (RT-PCR), miRNA RT-PCR, and western blotting were used to detect the expression levels of miR-503-5p and its targets in human MeT-5A mesothelial cells. A cell counting kit-8 assay was used to ascertain the impact of miR-503-5p on cellular proliferation, and cell cycle was evaluated using a FACScan flow cytometer. The miR-503-5p target genes were identified using mimic/inhibitor transfection and dual-luciferase reporter assays. The PDE exosomes (PDE-exos) labeled with PKH67 were used to observe their interactions with MeT-5A cells.</p> Results <p>Patients with PD who exhibited an increasing PSTR had notably elevated dialysate interleukin (IL)-6 concentrations, which demonstrated a positive correlation with the PSTR. The induction of lipopolysaccharides remarkably enhanced the expression of miR-503-5p and IL-6 in peritoneal mesothelial cells (PMCs). Overexpression of miR-503-5p induced cell cycle arrest in the G1/S phase and notably upregulated the expression of fibrotic indicators (alpha-smooth muscle actin and type I collagen) in PMCs. Cyclin D1 was a direct target of miR-503-5p and expressed in PMCs. Additionally, PKH67-labeled PDE-exos were internalized by PMCs.</p> Conclusions <p>Patients with PD who had a high PSTR exhibited an elevated inflammatory state, and the presence of inflammatory factors stimulated the upregulation of miR-503-5p expression, which led to cell cycle arrest in PMCs and promoted peritoneal fibrosis development.</p>

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MiR-503-5p mediates cell cycle arrest and fibrosis of peritoneal mesothelial cells with a high peritoneal solute transport status

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年11月24日

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  • Ouyang Ji,
  • Yingxin Xie,
  • Yunyao Lin,
  • Ruolin Li,
  • Yan Tong,
  • Pu Li,
  • Junyan Fang,
  • Yingli Liu

摘要

Background

Long-term peritoneal dialysis (PD) induces persistent inflammation and fibrosis of the peritoneal membrane (PM), altering the peritoneal solute transport rate (PSTR). We previously found elevated miR-503-5p levels in exosomes from PD effluent (PDE) in patients with high PSTR. As PSTR is strongly influenced by intraperitoneal inflammation, this study aimed to explore the relationship between miR-503-5p, inflammation, and peritoneal fibrosis and the underlying mechanisms.

Methods

Bioinformatics analyses were initially performed to identify the miR-503-5p target genes and determine the research subject. Real-time polymerase chain reaction (RT-PCR), miRNA RT-PCR, and western blotting were used to detect the expression levels of miR-503-5p and its targets in human MeT-5A mesothelial cells. A cell counting kit-8 assay was used to ascertain the impact of miR-503-5p on cellular proliferation, and cell cycle was evaluated using a FACScan flow cytometer. The miR-503-5p target genes were identified using mimic/inhibitor transfection and dual-luciferase reporter assays. The PDE exosomes (PDE-exos) labeled with PKH67 were used to observe their interactions with MeT-5A cells.

Results

Patients with PD who exhibited an increasing PSTR had notably elevated dialysate interleukin (IL)-6 concentrations, which demonstrated a positive correlation with the PSTR. The induction of lipopolysaccharides remarkably enhanced the expression of miR-503-5p and IL-6 in peritoneal mesothelial cells (PMCs). Overexpression of miR-503-5p induced cell cycle arrest in the G1/S phase and notably upregulated the expression of fibrotic indicators (alpha-smooth muscle actin and type I collagen) in PMCs. Cyclin D1 was a direct target of miR-503-5p and expressed in PMCs. Additionally, PKH67-labeled PDE-exos were internalized by PMCs.

Conclusions

Patients with PD who had a high PSTR exhibited an elevated inflammatory state, and the presence of inflammatory factors stimulated the upregulation of miR-503-5p expression, which led to cell cycle arrest in PMCs and promoted peritoneal fibrosis development.