Albendazole Reduces Vascular Smooth Muscle Cell Proliferation And Neointimal Formation Following Vascular Injury In Mice

Objective:Albendazole is an antihelminthic drug in parasites. To determin the utility of albendazole as a potential treatment for vascular disease involving proliferation and migration of human aortic smooth muscle cells, the effects of albendazole were tested in vitro and in a mouse model of arterial injury.Methods:Human aortic smooth muscle cells (HASMCs) were treated with DMSO or 0.25μM,0.5μM,1 μM albendazole (ABZ). The cell proliferation was detected by CCK-8 at 24,48 and 72 hours. The cellular migration was detected by cell wound scratch assay at 10 and 24 hours. The cellular invasion was detected by transwell assasy at 10 and 24 hours. Apoptosis was evaluated via TUNEL staining. The effect of ABZ on cell cycle distribution was observed by PI staining and Western blot assay was used to detect the expression of cell cycle related proteins and signal proteins. The effect of ABZ on tube-like strctures formation in human vein endothelial cells in vitro was observed by tube formation assay. Phenotypic switch-related mRNA and proteins were detected using real time PCR and Western blot. The expression of AKT、p-AKT、mTOR、p-mTOR、p-S6K at the protein level was examined at 24 hours after transfection. In vivo, femoral arterial wire injury was induced in wild-type mice treated with either albendazole or placebo control. HE staining and Evans blue dye were used to determin neointimal area and re-endothelization.Results:CCK8 results showed that ABZ significantly inhibited proliferation at 24,28 and 72h, respectively. ABZ inhibited migration of human aortic smooth muscle cells by cell wound scratch assay and transwell assay showed at 10 and 24h. The flow cytometry assay and Western blot showed that ABZ could obviously induced HASMCs cell cycle arrest at G2/M phase and markedly downregulated the expression of cell cycle related proteins CKD1 and cyclinBl proteins. The semi-quantitative RT-PCR and Western blot showed that phenotypic switch-related mRNA and proteins SMA、SM22a、MYH11 were dramatically increased after treatment of ABZ; ABZ down-regulated the PI3K/Akt/mTOR signal pathway. In femoral arterial wire injury, compared with placebo-treated mice, albendazole treated mice formed less neointimal at the site of injury by about 50%in mice after vascular injury. Re-endotheliazation was not affected by ABZ.Conclusion:1. ABZ could effectively inhibit human aortic smooth muscle cell proliferation and migration, this result may due to the arrest of cell cycle at G2/M phase and downregulation of cell cycle related proteins CKD1 and cyclinB1 proteins.2. ABZ significantly increased VSMC differentiation marker genes and down regulated the PI3K/Akt/mTOR signal pathway. ABZ decreased tube formation of HUVECs; however, ABZ had little effect on HUVECs proliferation and apoptosis of HASMCs.3. In vivo, ABZ inhibited neointimal area but dose not affect the process of re-endolization in mouse femoral arterial wire injury model, and ABZ may be a promising candidate of anti-restenosis drugs.