Curcumin Attenuates Ang-Ⅱ-induced VSMC Proliferation And Migration Via Downregulation Of PFKFB3

Objective:Vascular smooth muscle cells(VSMC)play a critical role in the pathogenesis of atherosclerosis and intimal hyperplasia.Recent studies highlight the importance of glucose metabolism in the control of VSMC proliferation and migration.This study investigates whether curcumin regulates angiotensin Ⅱ(Ang-Ⅱ)-induced VSMC proliferation and migration via phosphofructokinase-2 / fructose-2,6-bisphosphatase3(PFKFB3).Methods: First,the effect of Ang-Ⅱ-induced VSMC cell’s viability,migration and PFKFB3 protein expression was detected.Then,we used 3PO,the PFKFB3 protein specific inhibitor,acting on Ang-Ⅱ-induced VSMC and tested the expression of PFKFB3 protein,the changes of2,6-diphosphate fructose,lactic acid content and cell viability,survival rate and cell migration.Then,the expression of PFKFB3 protein was observed by using different concentrations of curcumin between normal VSMC and Ang-Ⅱ-induced VSMC.Finally,the concentration of curcumin was determined,and acting on Ang-Ⅱ-induced VSMC,then testing the expression of PFKFB3 protein,the changes of 2,6-diphosphate fructose,lactic acid content and cell viability,survival rate and cell migration.In this study,PFKFB3 protein level was detected by Western blot.The content of 2,6 diphosphate fructose and lactate production were detected by ELISA,cell viability was detected by cell counting kit-8,cell survival was detected by BrdU and cell migration distance was detected by wound-healing.Results: Exposure of VSMC to Ang-Ⅱ resulted in a significant increase in proliferation and migration.This was accompanied by an up-regulation of PFKFB3 expression and elevation of2,6 diphosphate fructose and lactate levels.Treatment with PFKFB3 inhibitor 3PO led to a significant decrease in Ang-Ⅱ induced VSMC proliferation and migration together with reduction of 2,6 diphosphate fructose,lactate production.Treatment of VSMC with curcumin blunted Ang-Ⅱ-induced the expression of PFKFB3 and productions of 2,6 diphosphate fructose and lactate in a dose-dependent manner.Our study demonstrates the importance of PFKFB3 in curcumin mediated reduction of the proliferation and migration of VSMC in response to Ang-Ⅱ.Conclusion: PFKFB3 plays a key role in the proliferation and migration of VSMC induced by Ang-Ⅱ Ⅱ and curcumin regulates Ang-Ⅱ induced proliferation and migration via PFKFB3 in VSMC.