ABSTRACT
Objective To investigate the effect and mechanism of chitosan on vascular smooth muscle cell proliferation of uremia patients with arteriovenous fistula.Methods Primarily culturing the VSMCs of uremia patients with arteriovenous fistula and patients without uremia by explants adherent method,and taking the second generation.VSMCs from patients without uremia cultured with 20％ FBS medium were non-uremia group,VSMCs of uremia patients cultured with 20％ FBS medium were uremia group,VSMCs of uremia patients with 100 pg/ml chitosan were uremia+ chitosan group.The expression of α-SMA was detected by immunohistochemistry.The changes of migration and invasion of VSMCs were detected by scratches and transwell migration assays.The mRNA expressions of TLR4 and PCNA were measured by real-time PCR.VSMCs of uremia patients with arteriovenous fistula were intervened with different doses of chitosan (0,100 and 500 μg/ml),and the protein expressions of TLR4,MyD88 and NF-κB were detected by Western blotting.Results Compared with those in non-uremia group,in uremia group and uremia+chitosan group α-SMA was upregulated,migration and invasion of VSMCs were enhanced,and mRNA expressions of TLR4 and PCNA were increased (all P ＜ 0.05).Compared with those in uremia group,the level of α-SMA was significantly decreased,the ability of migration and invasion of VSMCs were decreased,and the mRNA expressions of TLR4 and PCNA were decreased (all P ＜ 0.05).TLR4,MyD88 and NF-κB protein expressions were reduced in concentration-dependent manner by 100 and 500 μg/ml chitosan.Conclusions (1) In vitro,chitosan decreases the ability of migration and invasion of VSMCs of uremia patients with arteriovenous fistula.(2) Chitosan inhibits the proliferation of VSMCs,which may be relevant in the decreased expressions of TLR4,MyD88 and NF-κB.