Sulodexide inhibits neointimal hyperplasia of arteriovenous fistulas in rats through inactivation of YAP

Abstract

Objective To explore the role of sulodexide (SDX) in neointimal hyperplasia of arteriovenous fistulas (AVFs) in chronic kidney disease (CKD) rats and its possible mechanism. Methods A total of 18 male rats (weighing 300±50 g) were randomly and equally divided into AVF group, CKD+AVF group (CKD induction followed by AVF surgery and then gavaged with normal saline for 2 months), and CKD+AVF+SDX group[treated as in the CKD+AVF group but with 8 mg/(kg·d) SDX gavage].HE staining was used to observe the degree of neointimal hyperplasia.The expression of Hippo pathway related molecules, Yes-associated protein (YAP), pYAP and connective tissue growth factor (CTGF, YAP downstream target protein, one of mesenchymal marker) was detected by immunofluorescence assay.After human umbilical vein cell fusion EAHy926 cells were treated with 0, 2.5, 5, 10, 20 or 40 μg/mL SDX for 24 h, and with 2.5 μg/mL SDX for 24, 48 or 72 h, respectively, CCK-8 assay was used to measure cell survival rate.Moreover, the serum sample from CKD rat was used to treat EAHy926 cells, and then the cells were treated with SDX or YAP inhibitor verteporfin.The expression levels of YAP, pYAP, CTGF and endothelial cell marker CD31 were detected by Western blotting. Results HE staining and immunofluorescence assay showed that CKD rats had serious neointimal hyperplasia in AVFs (P < 0.05), and slightly lower expression of pYAP and enhanced expression of CTGF (P < 0.05) when compared with the rats of the AVF group.While, SDX treatment alleviated the neointimal hyperplasia of AVFs, enhanced the expression of pYAP and reduced the expression of CTGF (P < 0.05).CCK-8 assay showed that cell survival rate was decreased significantly in a dose-and time-dependent manner after SDX treatment (P < 0.05).Western blotting revealed that SDX increased the expression of pYAP and CD31 while inhibited the expression of CTGF in EAHy926 cells (P < 0.05), which was consistent with the effect of verteporfin treatment. Conclusion SDX can block YAP activation caused by CKD and attenuate neointimal hyperplasia in AVFs.

Keywords

• arteriovenous fistulas
• neointimal hyperplasia
• sulodexide
• yes-associated protein
• endothelial-mesenchymal transition