Abstract:

Objective To investigate the expression of a disintegrin and metalloproteinase 10 （ADAM10） in vascular tissue at the stenosis of human arteriovenous fistula （AVF） and its effect on the proliferation and migration of the vascular smooth muscle cells（VSMCs）， and to clarify its possible molecular mechanism. Methods The stenotic venous vascular tissue of 42 patients with end-stage renal disease （ESRD） who underwent reoperation due to AVF stenosis（AVF group） and their normal venous vascular tissue during the first operation （normal control group） were collected.Immunohistochemistry assay was used to detect the expressions of ADAM10 protein in venous vascular tissue of the patients in two groups；real-time fluorescence quantitative PCR（RT-qPCR） assay was used to detect the expression levels of ADAM10 mRNA in venous vascular tissue of the patients in two groups；the VSMCs were divided into control group and model group ［lipopoly saccharide（LPS） group，given 10 mg·L^－1 LPS］， LPS + si-NC group （transfected with si-NC plasmid +10 mg·L^－1LPS）， LPS+si-ADAM10 group （transfected with si-ADAM10 plasmid +10 mg·L^－1 LPS），LPS+Notch signal pathway inhibitor DAPT group （10 mg·L^－1 LPS+10 μmol·L^－1 Notch signal pathway inhibitor DAPT）， and LPS+si-ADAM10+DAPT group （transfected with si-ADAM10 plasmid +10 mg·L^－1 LPS+10 μmol·L^－1 DATP）； CCK-8 method was used to detect the proliferation activities of the cells in various groups； Transwell assay was used to detect the numbers of the migration cells in various groups； Western blotting method was used to detect the expression levels of ADAM10，proliferating cell nuclear antigen （PCNA），matrix metalloproteinase-9 （MMP-9）， Notch homolog protein 1 （Notch1）， Notch intracellular domain （NICD）， and hairy and enhancer of split 1 （Hes1） proteins in the cells in various groups. Results Compared with normal control group， the expression levels of ADAM10 protein and ADAM10 mRNA in venous vascular tissue of the patients in AVF group were significantly increased （P<0.05）. Compared with control group， the proliferation activity，number of the migration cells and expression levels of ADAM10， PCNA， MMP-9， Notch1， NICD， and Hes1 proteins in the cells in LPS group were significantly increased （P<0.05）； compared with LPS and LPS+si-NC group， the proliferation activity and number of the migration cells and expression levels of ADAM10， PCNA， MMP-9， Notch1， NICD， and Hes1 proteins in the cells in LPS+si-ADAM10 group were significantly decreased （P<0.05）. Compared with LPS group， the proliferative activities，numbers of the migration cells and expression levels of PCNA， MMP-9， Notch1， NICD， and Hes1 proteins in the cells in LPS+si-ADAM10 group and LPS+DAPI group were significantly decreased （P<0.05）； compared with LPS+si-ADAM10 group， the proliferation activity， number of the migration cells and expression levels of PCNA， MMP-9， Notch1， NICD， and Hes1 proteins in the cells in LPS+si-ADAM10+DAPT group were significantly decreased （P<0.05）. Conclusion ADAM10 is highly expressed in the vascular tissue at the stenosis of AVF，and silencing the expression of ADAM10 gene can inhibit the proliferation and migration of the VSMCs induced by LPS；its mechanism may be related to blocking the Notch signal pathway.

Key words: A disintegrin and metalloproteinase 10, Arteriovenous fistula, Vascular smooth muscle cells, Cell proliferation, Cell migration