Study On The Phenotypic Transformation Of Smooth Muscle Cells Induced By PDGF-BB And Its Mechanism By Total Extract Of Salvia Miltiorrhiza

Objective: the main function of vascular smooth muscle cells(VSMCs)is to contract and regulate the size and state of vascular diameter,the distribution of blood flow and blood pressure.When vascular injury or pathological stimulation occurs,VSMCs will undergo phenotypic transition.Under normal condition,VSMCs will change from contractile type to synthetic type,which is characterized by increased proliferation,migration and extracellular matrix synthesis,eventually leading to the occurrence of cardiovascular disease.Therefore,drugs targeting the phenotypic transition of vascular smooth muscle cells may be an effective treatment for the prevention and treatment of cardiovascular diseases.Danshen is a plant of Labiatae in the plant kingdom.It has many pharmacological activities,such as protecting vascular endothelial cells,improving coronary blood flow in myocardial ischemic area etc.It has a wide range of applications in cardiovascular diseases.The aim of this study was to investigate the effect and mechanism of total extract of Danshen on PDGF-BB-induced phenotypic transition of mouse vascular smooth muscle cells.Methods: With C57 BL / 6 mice of 6-8 weeks old,the smooth muscle cells were isolated by tissue attachment method,identified and subcultured Smooth muscle cells were identified by ?-SMA immunofluorescence,and 3-8 passages of vascular smooth muscle cells were taken for related experiments.The cultured vascular smooth muscle cells were divided into control group,PDGF-BB(10 ng/ml),low-dose Danshen group(P + DS 0.6 mg / mL),medium dose Danshen group(P + DS 0.8 mg /mL)and high-dose Danshen group(P + DS 1.0 mg / mL).CCK8 was used to detect the cell activity and proliferation rate;scratch test was used to detect the migration ability of vascular smooth muscle cells;Western blot was used to detect the protein expression of ?-SMA,OPN,IL-6,HIF-1 and MMP9.Results:(1)the purity of SMCs cultured to the third passage was more than 95%by inverted microscope and immunofluorescence detection.(2)Compared with PDGF-BB group,the cell proliferation rate and cell migration rate of low-dose Danshen group(P + DS 0.6 mg / mL),medium dose Danshen group(P + DS 0.8 mg /mL)and high-dose Danshen group(P + DS 1.0 mg / mL)were significantly decreased by a dose-dependent manner.(3)In PDGF-BB intervention group,the expression of OPN protein was significantly higher than that in the control group,and the expression of ?-SMA protein was significantly lower than that in the control group.After intervention with Danshen,the expression of ?-SMA induced by PDGF-BB was increased and the expression of OPN induced by PDGF-BB was decreased in a concentration dependent manner.(4)The expression of IL-6,HIF-1 and MMP9 protein in PDGF-BB intervention group was significantly higher than that in control group;Danshen reduced the expression of IL-6,HIF-1 and MMP9 protein induced by PDGF-BB in a concentration dependent manner;Conclusion:(1)PDGF-BB can promote the proliferation,migration and phenotype transition of primary VSMCs.(2)The total extract of Danshen could inhibit the proliferation,migration and phenotype transition of primary VCMCs induced by PDGF-BB.(3)The total extract of Danshen can inhibit the up regulation of HIF-1 protein expression in primary VCMCs induced by PDGF-BB,and inhibit the expression of IL-6 protein and MMP9 protein,so as to decrease the occurrence of cardiovascular disease.Innovation:(1)The biological functions and molecular pathways of PDGF-BB-induced phenotypic transformation of VSMCs have been studied.(2)through the WB experiment,the interaction between HIF-1,MMP9 and IL-6 proteins was detected.(3)the total extract of Salvia miltiorrhiza was used to inhibit the phenotypic transformation of VCMCs.