Effect Of Sulodexide On Peritoneal Morphology And Function Of Peritoneal Dialysis Rats

【Abstract】ObjectiveTo investigate the effect of sulodexide on peritoneal morphology and function of long-term peritoneal dialysis rats.MethodsThirty-six SD rats were randomly divided into four groups: control group (n=6) received no treatment; model group (n=10) received 20mL 4.25% peritoneal dialysis fluid daily IP for 8 weeks; low-dose SLX group (n=10) received 20mL 4.25% peritoneal dialysis fluid plus 10mg·kg^-1.BW SLX daily IP for 8 weeks; high-dose SLX group (n=10) received 20mL 4.25% peritoneal dialysis fluid plus 20mg·kg^-1·BW SLX daily IP for 8 weeks. Except for control group, peritoneal catheters were implanted in another thirty rats. At the end of the study, a 1 hour peritoneal equilibration test was performed , and leukocytes in dialysate samples were evaluated using hemocytometer. Then all the rats were humanely killed and blood clotting were estimated. The parietal peritoneum tissues of rats were harvested and stained by hematoxylin-eosin and Masson trichrome. Vessels and leukocytes of peritoneum tissue were counted with sections stained by HE and numbered as n/mm2. The levels of inflammatory cytokine (MCP-1, TNFα) in dialysate samples were measured by enzyme-linked immunosorbent assay(ELISA). Mesenteric expression ofα-SMA,TGFβ₁and CTGF mRNA was evaluated by RT-PCR.The morphology changes of partial peritoneum were examined by light microscopy and the expression ofα-SMA,TGFβ₁ and CTGF in parietal peritoneum were detected with immunohistochemistry assay.Results1. Nine rats were excluded from further analysis because of clogging of the catheter or peritonitis. There were no differences in body weight, prothrombin time, activated partial thromboplastin time,thrombin time and fibrinogen among the four groups(P>0.05).2. Compared with control group, the ultrafilitration volume (UF) and glucose reabsorption (D₁/D₀) were significantly lower in model group,and the dialysate- to-plasma urea ratio(D/Purea), dialysate -to-plasma total protein ratio(D/P_tp) and dialysate-to-plasma albumin ratio(D/P_alb) were significantly higher (P < 0.05). Administration of SLX can ameliorate peritoneal function significantly.3.Compared with control group, the number of peritoneal vessels, inflammatory cells, and peritoneal thickness were significantly increased. The number of leucocyte and the levels of MCP-1 and TNFαin 1 hour dialysate samples were also significantly increased(P<0.05).Additional use of sulodexide resulted in preserved the number of peritoneal vessels, inflammatory cells,and peritoneal thickness, and significantly decreased the number of leucocyte and the levels of MCP-1 and TNFαin 1 hour dialysate samples (P<0.05). But there was no significant difference between the low-dose SLX group and the high-dose SLX group(P>0.05).4.Compared with control group,α-SMA,TGFβ₁ and CTGF mRNA levels and the expression ofα-SMA,TGFβ₁ and CTGF in peritoneum were significantly increased in model group(P < 0.05). Up-regulation of the expression ofα-SMA,TGFβ₁ and CTGF mRNA was suppressed by SLX (P<0.05). No significant difference was observed between the low-dose SLX group and the high-dose SLX group (P>0.05).Conclusions1. A type of peritoneal fibrosis model in rat was successfully established .2. SLX could ameliorate peritoneal fibrosis and improve the structural and functional parameters of peritoneum.3. SLX could improve peritoneal fibrosis through inhibit pertioneal chronic inflammation, down-regulation of TGFβ₁, CTGF,and inhibit EMT of peritoneal mesothelial cells.