Calpain activation promotes dialysis-associated peritoneal fibrosis in rats

Abstract

To explore the role of calpain activation in the progression of peritoneal fibrosis. Twenty-four male Sprague-Dawley rats were randomized equally into control group, MDL28170 (a calpain inhibitor)+normal saline group, peritoneal dialysis (PD) model group and PD + MDL28170 group. In the latter two groups, the rats received daily intraperitoneal injections of 100 mL/kg of 4.25% glucose PD solution, and those in PD+MDL28170 group and MDL28170 saline group received daily infusion of 4 mg/kg MDL28170 every other day. Eight weeks later, the rats were euthanized for pathological examination of the parietal peritoneum, and the visceral peritoneum was used for examining the activation status of calpain and the expressions of fibronectin (FN) and collagen I (COL-I). Calpain activation and expressions of FN, COL-I and α-SMA were also examined using Western blotting and immunofluorescence assay in primary cultures of rat peritoneal mesothelial cells treated with MDL28170, transforming growth factor-β (TGF-β), or both. Compared with the control rats, the rats in PD model group showed significantly increased peritoneal peritoneum thickness, calpain activation in the peritoneal tissue, and expressions of FN and COL-I (P < 0.05). Treatment with MDL28170 significantly alleviated associated peritoneal fibrosis, decreased the thickness of the peritoneum (P < 0.05), and reduced the expressions of FN and COL-I in the rats with daily PD (P < 0.05). In the in vitro experiment, the expressions of FN and COL-I were also significantly lower in rat peritoneal mesothelial cells treated with both MDL28170 and TGF-β than in the cells treated with TGF-β alone (P < 0.05). Peritoneal calpain activity and expressions FN and COL-I all increase significantly in rat models of PD-associated peritoneal fibrosis. Calpain activation can promote peritoneal fibrosis, and inhibition of calpain can alleviate peritoneal fibrosis.