| Objective:To explore the effect of puerarin on neointimal formation in vivo and in vitro,and to elucidate its effect and possible mechanism on the proliferation,migration and phenotypic transformation of vascular smooth muscle cells.Content:The main reason for neointimal formation is the phenotypic transformation of vascular smooth muscle cells.Puerarin is the main component of Puerariae Lobatae Radix.In this study,the model of common carotid artery endothelial injury in mice and the model of phenotypic transformation of vascular smooth muscle cells in vitro were established to explore the effect of puerarin on neointimal formation and the effect and mechanism of phenotypic transformation of vascular smooth muscle cells.The etiology and pathogenesis of related diseases in Chinese and Western medicine were discussed,and the research progress of modern Chinese medicine on restenosis after intervention was reviewed.Methods:1.Effect of puerarin on neointima after common carotid artery endothelial injury in mice: Male B129 mice were randomly divided into Control group,Sham group,Model group and Puerarin group,with 10 mice in each group.The Control group and Sham group were performed with an incision on the left side of the neck,while the Model group and Puerarin group were performed with common carotid artery endothelial injury.After 4 weeks of intragastric administration of normal saline or puerarin,the common carotid artery was taken out and embedded with OCT for HE staining,and the intima/media area ratio was analyzed and calculated,and the expression of OPN was detected by immunofluorescence.2.The effect of puerarin on the proliferation,migration and phenotypic transformation of vascular smooth muscle cells : vascular smooth muscle cells were extracted from SD rats and cultured to the third generation to verify their cell purity.PDGF-BB was used to induce vascular smooth muscle cells to transform from contractile phenotype to synthetic phenotype in vitro.CCK8 was used to detect drug toxicity and cell proliferation.Scratch test and transwell chamber were used to detect cell migration.Western blot was used to detect the protein expression of contractile phenotype markers SMMHC,smoothelin,calponin and synthetic phenotype marker OPN.3.The mechanism of puerarin on phenotypic transformation of vascular smooth muscle cells : Western blot was used to detect the expression of p38 MAPK and its phosphorylated protein in vascular smooth muscle cells treated with puerarin and PDGF-BB.Results:1.Puerarin can inhibit neointimal formation after endothelial injury of common carotid artery in mice: After endothelial injury of common carotid artery in mice,the lumen area decreased and the intima / media area increased,indicating that the model was successfully established.Compared with the Model group,the intima / media area of the Puerarin group was significantly reduced,and the expression of OPN in the neointima was also reduced.2.Puerarin can inhibit the proliferation,migration and phenotypic transformation of vascular smooth muscle cells cultured in vitro : Puerarin can inhibit the proliferation of vascular smooth muscle cells induced by PDGF-BB,and the inhibition is dose-dependent.Puerarin could inhibit PDGF-BB-induced vascular smooth muscle cell migration in a dose-dependent manner.PDGF-BB can increase the expression of OPN in vascular smooth muscle cells,and decrease the expression of SMMHC,smoothelin and calponin.Puerarin can reverse these results.3.Puerarin inhibits the phenotypic transformation of vascular smooth muscle cells by regulating p38 MAPK pathway : Compared with the Control group,the expression of p38 MAPK phosphorylation in PDGF-BB group was up-regulated.After puerarin intervention,the expression of p38 MAPK phosphorylation was down-regulated compared with PDGF-BB group,and the expression levels of p38 MAPK in the three groups were unchanged.Conclusion:1.Puerarin can inhibit neointimal formation after carotid artery endothelial injury in mice,which is mainly related to the inhibition of vascular smooth muscle cells phenotypic transformation,suggesting that puerarin has a certain preventive effect on restenosis after PCI.2.Puerarin inhibits PDGF-BB-induced phenotypic transformation of vascular smooth muscle cells by inhibiting p38 MAPK pathway. |
