The Effects Of Febuxostat On VSMC Proliferation And Migration In Mouse

Background:The abnormal proliferation and migration of vascular smooth muscle cells (VSMC) play an important role in the development of atherosclerosis lesions. reactive oxygen species(ROS) is a key factor to stimulate the smooth muscle cell proliferation and migration. Febuxostat,a new type of xanthine oxidase inhibitors,can reduce the production of ROS by inhibiting the activity of xanthine oxidase. However, the effects of febuxostat in the regulation platelet derived growth factor(PDGF) induced VSMC proliferation and migration has not been identified.Objective:In this study, we investigate the effect of febuxostat on PDGF-BB-induced VSMC proliferation and migration.Method:Male SD rats weighted between 180 and 200 g were selected, the thoracic aortas of rats were isolated, vascular smooth muscle cells between the fourth and the fifth generation were extracted by explant culture. These cells can be divided into the following four groups:control、PDGF、PDGF+XO、PDGF+XO+Fx, PDGF +XO+Fx group can also be divided into PDGF+XO+Fx(1μM)、 PDGF+XO+ Fx(5μM) and PDGF+XO+Fx(10μM) according to the different concentrations of Fx. PDGF+XO group and PDGF+XO+Fx group were incubated with culture medium including the same concentration of XO(5mU/ml) and the different concentrations of Fx(1μM、5μM、10μM) for 24h. After 24h,the culture medium were replaced,and cells were incubated with culture medium including the same concentration of PDGF(20ng/ml) for 24h.CCK-8 method was used to detect the effect of VSMC proliferation, transwell method was used to detect the effect of VSMC migration, fluid cytology and Western Blot method was used to detect the effect of VSMC cell cycle, Western Blot method wes used to detect the effect of PDGF signaling pathway.Result:1. After giving stimulation with PDGF and XO, cell proliferation was significantly enhanced in PDGF+XO (0.964±0.049) group compared with control group(0.538±0.074), P<0.05; after giving stimulation with different concentration of Fx (1μM、5μM、10μM、20μM), cell proliferation was gradually weakened in a dose-dependent manner, cell proliferation was significantly weakened in PDGF+XO +Fx(10μM)(0.573±0.055) group compared with PDGF+XO group, P<0.05.2. the cells that migrated through the polycarbonate membrane were significantly increased in PDGF+XO(37.6±4.8)group compared with control group(9.6±2.2), P<0.05; after giving stimulation with Fx, the cells were significantly decreased in PDGF+XO+Fx (14.4±2.1) group compared with PDGF+XO group, P<0.05.3. After giving stimulation with PDGF and XO, the percent of S-phase(19.70±1.21%)of cell cycle in PDGF+XO group was higher than that in control group (8.49±1.50%),P<0.05; after giving stimulation with Fx, the percent (12.10±1.73%) in PDGF+XO+Fx was lower than that in PDGF+XO group (19.70±1.21%), P<0.05.4. After giving stimulation with PDGF and XO, the expression of cell cycle regulation proteins (cyclin D1,CDK4, cyclin E and CDK2) in PDGF+XO group was higher than that in control group, however, this phenomenon can be inhibited by Fx in a dose-dependent manner.5. febuxostat potently inhibited the increase of pAkt and pERK1/2 induced by PDGF in a dose-dependent manner.Conclusion:1.febuxostat significantly inhibited VSMC proliferation and migration induced by PDGF.2.The inhibition of VSMC proliferation by febuxostat was associated with cell cycle.3.febuxostat potently inhibited the activation of Akt and ERK signaling pathway induced by PDGF.