Effect of ulinastatin on proliferation, migration and apoptosis of vascular smooth muscle cells in human aortic dissection by regulating GSK-3β/β-catenin signaling pathway

AIM To investigate the effect of ulinastatin (UTI) on the proliferation, migration and apoptosis of vascular smooth muscle cells (VSMCs) in human aortic dissection (AD) by regulating the glycogen synthase kinase-3β (GSK-3β)/β-catenin signaling pathway.

METHODS Primary VSMCs were extracted from the excised blood vessels of AD patients using tissue block adhesion method and were passaged for culture. Coomassie Brilliant Blue staining and α-SMA immunofluorescence staining were performed to identify cells. VSMCs cultured in vitro were randomly grouped into Control group, L-UTI group (500 μmol/L UTI), H-UTI group (1500 μmol/L UTI), H-UTI+pcDNA-NC group (1500 μmol/L UTI+transfected with pcDNA-NC) and H-UTI+pcDNA-SK-3β group (1500 μmol/L UTI+transfected with pcDNA-SK-3β). QRT-PCR was applied to detect the relative expression level of GSK-3β of cells in each group, CCK8 and plate clone formation experiments were performed to detect the proliferation of cells in each group, scratch experiment was conducted to detect the migration of cells in each group, flow cytometry was applied to detect apoptosis of cells in various groups and Western blot was used to detect the expression of proliferating cell nuclear antigen (PCNA), B cell-lymphotumor-2 (Bcl-2), Bcl-2 associated X protein (Bax), and GSK-3β/S1P/S1PR3 signaling pathway related proteins (p-GSK-3β/GSK-3β, β-catenin) of cells in each group.

RESULTS Compared with those in Control group, the survival rate, clone number, scratch healing rate, PCNA, Bcl-2, p-GSK-3β/GSK-3β and β-catenin protein expressions were lower in L-UTI group and H-UTI group, while the apoptosis rate and Bax protein expression were higher (P<0.05, P<0.01). Compared with those in H-UTI group or H-UTI+pcDNA-NC group, the survival rate, clone number, scratch healing rate, PCNA, Bcl-2, p-GSK-3β/GSK-3β and β-catenin protein expressions of cells were higher in H-UTI+pcDNA-SK-3β group, while the apoptosis rate and Bax protein expression were lower (all P<0.05).

CONCLUSION UTI may induce VSMC apoptosis and inhibit cell proliferation and migration in AD patients by regulating the GSK-3β/β-catenin signaling pathway.